Journal: Biomolecules

Article Title: Transcriptome Analyses Identify Deregulated MYC in Early Onset Colorectal Cancer

doi: 10.3390/biom12091223

Figure Lengend Snippet: MYC and PVT1 copy number alterations are found in some patient tumors. ( A ) Bar graph depicting the copy number of the MYC gene locus in patient-matched uninvolved colonic segments (normal, N) and tumors (T). Highlighted in blue are tumor samples with 2.5 or greater copy number. ( B ) Correlation of MYC copy number and MYC expression in tumors. Red points are tumors expressing high- MYC and blue points are tumors expressing low- MYC (Pearson correlation estimate 0.68, p = 5.45 × 10 −6 ). ( C ) Bar graph depicting the copy number of the PVT1 locus in the six patient-matched samples displaying gains in the MYC genomic locus in tumors (T). Uninvolved colonic tissues (normal, N) are shown as a reference. In ( A , C ), highlighted in blue are tumors samples with 2.5 or greater increases in copy number. Error bars represent the range of copy numbers detected in four technical replicates per sample.

Article Snippet: The MYC genomic locus was evaluated using the Taqman probe Hs02602824_cn (Thermo Fisher), and the PVT1 genomic locus was evaluated using Taqman probe Hs06232749_cn.

Techniques: Expressing

Journal: Biomolecules

Article Title: Transcriptome Analyses Identify Deregulated MYC in Early Onset Colorectal Cancer

doi: 10.3390/biom12091223

Figure Lengend Snippet: Expression of PVT1 correlates with MYC expression. ( A ) rlog normalized expression plot of PVT1 in patient-matched adjacent colonic tissues (Normal) and tumors (paired Wilcoxon test, p = 4.8 × 10 −6 ). Each point represents a single patient with a line drawn between matched samples. ( B ) Correlation plot depicting normalized MYC versus normalized PVT1 gene expression in adjacent colonic segments (Normal, blue points) and tumors (red points). ( C ) Correlation plot depicting PVT1 and MYC differential expression in patient-matched adjacent colonic segments and tumors.

Article Snippet: The MYC genomic locus was evaluated using the Taqman probe Hs02602824_cn (Thermo Fisher), and the PVT1 genomic locus was evaluated using Taqman probe Hs06232749_cn.

Techniques: Expressing, Gene Expression, Quantitative Proteomics

Journal: BioMed Research International

Article Title: Association of lncRNA PVT1 Gene Polymorphisms with the Risk of Essential Hypertension in Chinese Population

doi: 10.1155/2022/9976909

Figure Lengend Snippet: Test results of Hardy-Weinberg Equilibrium among all PVT1 gene sites.

Article Snippet: Assay ID for rs10956390 is C__317048_10 (Thermo), and assay ID for rs80177647 is C__101092625_10 (Thermo).

Techniques: Control

Journal: BioMed Research International

Article Title: Association of lncRNA PVT1 Gene Polymorphisms with the Risk of Essential Hypertension in Chinese Population

doi: 10.1155/2022/9976909

Figure Lengend Snippet: Association between lncRNA PVT1 polymorphisms and EH risk in the south of China.

Article Snippet: Assay ID for rs10956390 is C__317048_10 (Thermo), and assay ID for rs80177647 is C__101092625_10 (Thermo).

Techniques: Control

Journal: BioMed Research International

Article Title: Association of lncRNA PVT1 Gene Polymorphisms with the Risk of Essential Hypertension in Chinese Population

doi: 10.1155/2022/9976909

Figure Lengend Snippet: Association between lncRNA PVT1 polymorphisms and EH risk in the north of China.

Article Snippet: Assay ID for rs10956390 is C__317048_10 (Thermo), and assay ID for rs80177647 is C__101092625_10 (Thermo).

Techniques: Control

Journal: BioMed Research International

Article Title: Association of lncRNA PVT1 Gene Polymorphisms with the Risk of Essential Hypertension in Chinese Population

doi: 10.1155/2022/9976909

Figure Lengend Snippet: Association between lncRNA PVT1 polymorphisms and EH risk in China.

Article Snippet: Assay ID for rs10956390 is C__317048_10 (Thermo), and assay ID for rs80177647 is C__101092625_10 (Thermo).

Techniques: Control

Journal: BioMed Research International

Article Title: Association of lncRNA PVT1 Gene Polymorphisms with the Risk of Essential Hypertension in Chinese Population

doi: 10.1155/2022/9976909

Figure Lengend Snippet: Influence of PVT1 rs10956390 polymorphism on lipid level of subjects: (a) TG; (b) TC; (c) HDL-C; (d) LDL-C. (Data are expressed as the mean ± SEM. Control: CC, n = 108, CT, n = 220, TT, n = 111; EH: CC, n = 192, CT, n = 233, TT, n = 99. ∗ P < 0.05, ∗∗ P < 0.01, as compared with CC genotype in interclass; # P < 0.05, as compared with CT genotype in interclass).

Article Snippet: Assay ID for rs10956390 is C__317048_10 (Thermo), and assay ID for rs80177647 is C__101092625_10 (Thermo).

Techniques: Control

Journal: BioMed Research International

Article Title: Association of lncRNA PVT1 Gene Polymorphisms with the Risk of Essential Hypertension in Chinese Population

doi: 10.1155/2022/9976909

Figure Lengend Snippet: Influence of PVT1 rs80177647 polymorphism on lipid level of subjects: (a) TG; (b) TC; (c) HDL-C; (d) LDL-C. (Data are expressed as the mean ± SEM. Control: TT, n = 377, TA, n = 57, AA, n = 5; EH: TT, n = 395, TA, n = 119, AA, n = 10.).

Article Snippet: Assay ID for rs10956390 is C__317048_10 (Thermo), and assay ID for rs80177647 is C__101092625_10 (Thermo).

Techniques: Control

Journal: BioMed Research International

Article Title: Association of lncRNA PVT1 Gene Polymorphisms with the Risk of Essential Hypertension in Chinese Population

doi: 10.1155/2022/9976909

Figure Lengend Snippet: Effect of PVT1 mRNA expression in whole blood from healthy normal donors: (a) rs10956390; (b) rs80177647.

Article Snippet: Assay ID for rs10956390 is C__317048_10 (Thermo), and assay ID for rs80177647 is C__101092625_10 (Thermo).

Techniques: Expressing

Journal: RSC Advances

Article Title: Retracted Article: PVT1 knockdown alleviates vancomycin-induced acute kidney injury by targeting miR-124 via inactivation of NF-κB signaling

doi: 10.1039/c8ra05724a

Figure Lengend Snippet: PVT1 was highly expressed in the kidneys of VAN-induced AKI mice. (A)–(E) Mice were intraperitoneally injected with VAN (600 mg kg −1 ) for the indicated time points (0, 1, 3, 7 day). (A) and (B) BUN and Scr levels were determined by respective commercial kits. (C) Histopathological alterations of kidney tissues were monitored by H&E staining. (D) TUNEL assay was performed to detect cell apoptosis pattern in kidney tissues. (E) Western blot assay was conducted to detect protein levels of Bax and Bcl-2 in kidney tissues. (F) PVT1 level in kidney tissues was measured by RT-qPCR assay. * P < 0.05.

Article Snippet: Then, the relative expression of PVT1 (mouse or human) was measured using TaqMan Gene Expression Assays (Applied Biosystems, Foster City, CA, USA) for PVT1 (Hs01069023_m1 or Mm01203908_m1) and GAPDH (Hs00266705_g1 or Mm99999915_g1) with GAPDH as the endogenous control.

Techniques: Injection, Staining, TUNEL Assay, Western Blot, Quantitative RT-PCR

Journal: RSC Advances

Article Title: Retracted Article: PVT1 knockdown alleviates vancomycin-induced acute kidney injury by targeting miR-124 via inactivation of NF-κB signaling

doi: 10.1039/c8ra05724a

Figure Lengend Snippet: PVT1 knockdown ameliorated VAN-induced AKI in vivo . (A)–(E) Mice were injected with si-NC or si-PVT1 for 24 h and then intraperitoneally injected with VAN (600 mg kg −1 ) for 7 days. (A) PVT1 level in kidney tissues were measured by RT-qPCR assay. (B) and (C) BUN and Scr levels were determined by matching commercial kits. (D) Histopathological changes of kidney tissues were detected by H&E staining. (E) Cell apoptosis in kidney tissues was tested by TUNEL assay. (F) Western blot assay was conducted to detect protein levels of Bax and Bcl-2 in kidney tissues. * P < 0.05.

Article Snippet: Then, the relative expression of PVT1 (mouse or human) was measured using TaqMan Gene Expression Assays (Applied Biosystems, Foster City, CA, USA) for PVT1 (Hs01069023_m1 or Mm01203908_m1) and GAPDH (Hs00266705_g1 or Mm99999915_g1) with GAPDH as the endogenous control.

Techniques: Knockdown, In Vivo, Injection, Quantitative RT-PCR, Staining, TUNEL Assay, Western Blot

Journal: RSC Advances

Article Title: Retracted Article: PVT1 knockdown alleviates vancomycin-induced acute kidney injury by targeting miR-124 via inactivation of NF-κB signaling

doi: 10.1039/c8ra05724a

Figure Lengend Snippet: PVT1 directly interacted with miR-124. (A) Predicted binding sites between PVT1 and miR-124 by miRcode online website and mutant sites in PVT1-MUT reporter. (B) 293T cells were co-transfected with miR-NC or miR-124 and PVT1-WT reporter or PVT1-MUT reporter, followed by the determination of luciferase activities at 48 h after transfection. (C) RIP and RT-qPCR assays were carried out to measure the enrichment levels of PVT1 and miR-124 in IgG or Ago2 immunoprecipitation complex in HK-2 cells. (D) RNA pull down and RT-qPCR assays were performed to investigate whether miR-124 could interact with PVT1 by predicted binding sites in HK-2 cells. The experiments were repeated at least three times. * P < 0.05.

Article Snippet: Then, the relative expression of PVT1 (mouse or human) was measured using TaqMan Gene Expression Assays (Applied Biosystems, Foster City, CA, USA) for PVT1 (Hs01069023_m1 or Mm01203908_m1) and GAPDH (Hs00266705_g1 or Mm99999915_g1) with GAPDH as the endogenous control.

Techniques: Binding Assay, Mutagenesis, Transfection, Luciferase, Quantitative RT-PCR, Immunoprecipitation

Journal: RSC Advances

Article Title: Retracted Article: PVT1 knockdown alleviates vancomycin-induced acute kidney injury by targeting miR-124 via inactivation of NF-κB signaling

doi: 10.1039/c8ra05724a

Figure Lengend Snippet: The silencing of PVT1 abated VAN-induced HK-2 cell apoptosis in vitro . (A)–(C) HK-2 cells were stimulated with 4 mM VAN for 24 h. (A) PVT1 level was measured by RT-qPCR assay. (B) Apoptosis index was determined by double-staining of Annexin V-FITC and PI using a flow cytometry. (C) Protein levels of Bcl-2 and Bax were detected by western blot assay. (D)–(F) HK-2 cells were transfected with si-NC or si-PVT1 for 24 h and then stimulated with 4 mM VAN for additional 24 h, followed by the determination of PVT1 (D), Bcl-2 (F) and Bax (F) levels, and apoptosis index (E). The experiments were repeated at least three times. * P < 0.05.

Article Snippet: Then, the relative expression of PVT1 (mouse or human) was measured using TaqMan Gene Expression Assays (Applied Biosystems, Foster City, CA, USA) for PVT1 (Hs01069023_m1 or Mm01203908_m1) and GAPDH (Hs00266705_g1 or Mm99999915_g1) with GAPDH as the endogenous control.

Techniques: In Vitro, Quantitative RT-PCR, Double Staining, Flow Cytometry, Western Blot, Transfection

Journal: RSC Advances

Article Title: Retracted Article: PVT1 knockdown alleviates vancomycin-induced acute kidney injury by targeting miR-124 via inactivation of NF-κB signaling

doi: 10.1039/c8ra05724a

Figure Lengend Snippet: The inhibition of miR-124 abrogated si-PVT1-mediated anti-apoptosis effect in VAN-treated HK-2 cells. (A) HK-2 cells were stimulated with 4 mM VAN for 24 h, followed by the measurement of miR-124 level via RT-qPCR assay. (B)–(D) HK-2 cells were transfected with si-NC, si-PVT1, si-PVT1 + anti-miR-NC or si-PVT1 + anti-miR-124 for 24 h and then treated with 4 mM VAN for 24 h followed by the measurement of miR-124 (B), Bcl-2 (D) and Bax (D) levels, and apoptosis index (C). The experiments were repeated at least three times. * P < 0.05.

Article Snippet: Then, the relative expression of PVT1 (mouse or human) was measured using TaqMan Gene Expression Assays (Applied Biosystems, Foster City, CA, USA) for PVT1 (Hs01069023_m1 or Mm01203908_m1) and GAPDH (Hs00266705_g1 or Mm99999915_g1) with GAPDH as the endogenous control.

Techniques: Inhibition, Quantitative RT-PCR, Transfection

Journal: RSC Advances

Article Title: Retracted Article: PVT1 knockdown alleviates vancomycin-induced acute kidney injury by targeting miR-124 via inactivation of NF-κB signaling

doi: 10.1039/c8ra05724a

Figure Lengend Snippet: PVT1 activated NF-κB signaling by targeting miR-124 in VAN-induced HK-2 cells. (A) and (B) HK-2 cells were transfected with pcDNA, PVT1, PVT1 + miR-NC, PVT1 + miR-124, si-NC, si-PVT1, si-PVT1 + anti-miR-NC, or si-PVT1 + anti-miR-124 for 24 h and then treated with 4 mM VAN for 24 h. Then, protein levels of p-p65, p65, p-IκBα and IκBα were determined by western blot assay. The experiments were repeated at least three times.* P < 0.05.

Article Snippet: Then, the relative expression of PVT1 (mouse or human) was measured using TaqMan Gene Expression Assays (Applied Biosystems, Foster City, CA, USA) for PVT1 (Hs01069023_m1 or Mm01203908_m1) and GAPDH (Hs00266705_g1 or Mm99999915_g1) with GAPDH as the endogenous control.

Techniques: Transfection, Western Blot

Journal: RSC Advances

Article Title: Retracted Article: PVT1 knockdown alleviates vancomycin-induced acute kidney injury by targeting miR-124 via inactivation of NF-κB signaling

doi: 10.1039/c8ra05724a

Figure Lengend Snippet: The schematic diagram of PVT1 knockdown alleviates vancomycin-induced acute kidney injury by targeting miR-124 via inactivating NF-κB signaling.

Article Snippet: Then, the relative expression of PVT1 (mouse or human) was measured using TaqMan Gene Expression Assays (Applied Biosystems, Foster City, CA, USA) for PVT1 (Hs01069023_m1 or Mm01203908_m1) and GAPDH (Hs00266705_g1 or Mm99999915_g1) with GAPDH as the endogenous control.

Techniques: Knockdown

Journal: Bioscience Reports

Article Title: Long non-coding RNA PVT1, a molecular sponge for miR-149, contributes aberrant metabolic dysfunction and inflammation in IL-1β-simulated osteoarthritic chondrocytes

doi: 10.1042/BSR20180576

Figure Lengend Snippet: ( A ) The expression levels of PVT1 in human OA tissues were determined by qRT-PCR. ( B ) After exposure to IL-1β (10 ng/ml) for 24 h, the expression of PVT1 in OA chondrocytes was also detected. * P <0.05 vs. control group.

Article Snippet: The biotinylated DNA probe complementary to PVT1 RNA was synthesized by GenePharma.

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Bioscience Reports

Article Title: Long non-coding RNA PVT1, a molecular sponge for miR-149, contributes aberrant metabolic dysfunction and inflammation in IL-1β-simulated osteoarthritic chondrocytes

doi: 10.1042/BSR20180576

Figure Lengend Snippet: ( A ) Cells were transfected with si-PVT1 or si-NC for 48 h. The expression of PVT1 was determined. * P <0.05 vs. si-NC-treated group. ( B ) After transfection with si-PVT1, cells were exposed to IL-1β. Then, the mRNA levels of aggrecan and collagen II were measured by qRT-PCR. ( C ) The corresponding protein levels of aggrecan and collagen II were analyzed by Western blotting. ( D ) Quantified analysis of protein was performed by Quantity One software. ( E ) The subsequent transcript levels of MMP-3, MMP-9 and MMP-13 were determined by qRT-PCR. ( F – H ) The contents of MMP-3 (F), MMP-9 (G) and MMP-13 (H) in supernatants were evaluated by ELISA assay. * P <0.05 vs. control group. # P <0.05 vs. IL-1β-treated group.

Article Snippet: The biotinylated DNA probe complementary to PVT1 RNA was synthesized by GenePharma.

Techniques: Transfection, Expressing, Quantitative RT-PCR, Western Blot, Software, Enzyme-linked Immunosorbent Assay, Control

Journal: Bioscience Reports

Article Title: Long non-coding RNA PVT1, a molecular sponge for miR-149, contributes aberrant metabolic dysfunction and inflammation in IL-1β-simulated osteoarthritic chondrocytes

doi: 10.1042/BSR20180576

Figure Lengend Snippet: ( A ) Chondrocytes were transfected with si-PVT1, prior to exposure to IL-1β for 24 h. Then, the concentration of PGE2 in supernatants was detected by ELISA. ( B ) The production of NO was analyzed by a NO detection kit. ( C – E ) The inflammatory cytokine levels of IL-6 (C), IL-8 (D) and TNF-α (E) in culture medium were also measured by ELISA. * P <0.05 vs. control group. # P <0.05 vs. IL-1β-treated group.

Article Snippet: The biotinylated DNA probe complementary to PVT1 RNA was synthesized by GenePharma.

Techniques: Transfection, Concentration Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: Bioscience Reports

Article Title: Long non-coding RNA PVT1, a molecular sponge for miR-149, contributes aberrant metabolic dysfunction and inflammation in IL-1β-simulated osteoarthritic chondrocytes

doi: 10.1042/BSR20180576

Figure Lengend Snippet: ( A ) Following transfection with si-PVT1, the expression of miR-149 was assessed by qRT-PCR. * P <0.05 vs. si-NC-treated group. ( B ) Cells were transfected with recombinant PVT1 (rPVT1) plasmids, then the expression of PVT1 was detected. ( C ) The expression of miR-149 in PVT1-overexpressed cells. * P <0.05 vs. vector-treated group. ( D ) Cells were treated with si-PVT1 or si-NC, and then were transfected with miR-149 sensor. After that, the luciferase activity was measured. ( E ) Schematic illustration of the presumed site complementary to miR-149. ( F ) After co-transection with Luc- PVT1-wt, or Lcu-PVT1-mut, and miR-149 mimics. Luciferase activity was then assessed. ( G ) miR-149 was pulled down using biotinylated PVT1 probe or random probe. Then, the expression levels of miR-149 were detected by Northern blot. ( H ) PVT1 is associated with miR-149. Cells were transfected with biotinylated miR-149 wt (Bio-149-wt) or its mutant (Bio-149-mut). The negative control (Bio-NC) means that a biotinylated microRNA that is not complementary to PVT1. Approximately 48 h later, the expression of PVT1 was evaluated by qRT-PCR; * P <0.05, ** P <0.01.

Article Snippet: The biotinylated DNA probe complementary to PVT1 RNA was synthesized by GenePharma.

Techniques: Transfection, Expressing, Quantitative RT-PCR, Recombinant, Plasmid Preparation, Luciferase, Activity Assay, Northern Blot, Mutagenesis, Negative Control

Journal: Bioscience Reports

Article Title: Long non-coding RNA PVT1, a molecular sponge for miR-149, contributes aberrant metabolic dysfunction and inflammation in IL-1β-simulated osteoarthritic chondrocytes

doi: 10.1042/BSR20180576

Figure Lengend Snippet: ( A ) The transfected efficiency of miR-149 inhibitor. ( B ) Cells were transfected with si-PVT1 and anti-miR-149 inhibitor, following the exposure to IL-1β. Then, the mRNA levels of aggrecan and collagen II were determined by qRT-PCR. ( C ) The corresponding effects on the transcripts of MMPs. ( D – I ) The subsequent levels of MMP-3 and MMP-13 (D), MMP-9 (E), PGE2 (F), NO (G), IL-6 (H), IL-8 and TNF-α (I) in culture supernatants were also detected. * P <0.05 vs. control group. # P <0.05 vs. si-PVT1 group.

Article Snippet: The biotinylated DNA probe complementary to PVT1 RNA was synthesized by GenePharma.

Techniques: Transfection, Quantitative RT-PCR, Control

Journal: Molecular Cancer

Article Title: Long noncoding RNA Pvt1 regulates the immunosuppression activity of granulocytic myeloid-derived suppressor cells in tumor-bearing mice

doi: 10.1186/s12943-019-0978-2

Figure Lengend Snippet: Pvt1 is highly expressed in tumor-expanded G-MDSCs. A total of 2 × 10 ^6 Lewis lung carcinoma cells (LLCs) were introduced via s.c. injection into C57BL/6 mice. After 4 weeks, bone marrow cells, splenocytes and a single-cell suspension derived from tumor tissues were collected, and G-MDSCs were later sorted. Splenocytes from wild-type (WT) C57BL/6 mice were collected, and G-MDSCs were isolated. Hierarchical clustering analysis of lncRNAs and protein-coding RNAs that were differentially expressed (fold change > 2) in G-MDSCs sorted from tumor tissue of Lewis tumor-bearing mice and spleens of WT C57BL/6 mice. a Clustering tree for lncRNAs; the expression values are represented in shades of red and green, indicating expression above and below normal values, respectively. b The purity of sorted G-MDSCs was determined via flow cytometry by assessing the expression of two surface markers: Ly6G and CD11b. c The expression level of Pvt1 in total RNA isolated from G-MDSCs from the bone marrow, spleen and tumor tissues of Lewis-bearing mice was measured by qRT-PCR. Fresh G-MDSCs isolated from bone marrow (BM) from WT C57BL/6 mice served as the control. Bone marrow cells (1 × 10 ^6 ) from WT C57BL/6 mice were plated in 24-well plates in 1 mL of RPMI 1640 medium containing 10% FBS, 20 ng/mL IL-6 and 20 ng/mL GM-CSF. The cells were then collected, and G-MDSCs were sorted 3 days later. d G-MDSCs cocultured with CFSE-labeled CD4 + T cells at a ratio of 1:1 in the presence of anti-CD3 mAb and anti-CD28 mAb for 72 h. The proliferation of CD4 + T cells was detected by flow cytometry at 488 nm excitation light. e Arg1 activity in G-MDSCs induced from BM cells was measured. f ROS production in G-MDSCs was analyzed via flow cytometry. g The expression level of Pvt1 in G-MDSCs was detected using qRT-PCR. *** p < 0.001, and ** p < 0.01; ns: no significance

Article Snippet: G-MDSCs were plated in 48-well plates or 24-well plates (Costar, Corning, NY) with RPMI 1640 medium supplemented with 10% fetal calf serum and then transfected with 50 nM Pvt1 siRNA or the negative control (Ribobio Co., Guangzhou, China) using Entranster-R (Engreen Biosystem Co., Beijing, China) according to the manufacturer’s instructions.

Techniques: Injection, Suspension, Derivative Assay, Isolation, Expressing, Flow Cytometry, Quantitative RT-PCR, Control, Labeling, Activity Assay

Journal: Molecular Cancer

Article Title: Long noncoding RNA Pvt1 regulates the immunosuppression activity of granulocytic myeloid-derived suppressor cells in tumor-bearing mice

doi: 10.1186/s12943-019-0978-2

Figure Lengend Snippet: Knockdown of Pvt1 alters the suppressive capacity of G-MDSCs in vitro. G-MDSCs sorted from tumor tissues were obtained from TB mice injected with cells transfected with 50 nM Pvt1 siRNA (si-Pvt1) or negative control (NC) siRNA (si-NC). a qRT-PCR confirmed the efficiency of transfection with si-Pvt1. b G-MDSCs were transfected with Pvt1 siRNA, and then, the cells were harvested after 6 h and cocultured with CD4 + T cells at ratio of 1:1 in the presence of anti-CD3 mAb and anti-CD28 mAb for72 h. 3H-thymidine incorporation was used to detect T cells proliferation. c Arg1 activity in G-MDSCs transfected with si-Pvt1 was measured. d ROS production in G-MDSCs was analyzed via flow cytometry. ** p < 0.01, and * p < 0.05; ns: no significance; Geo MFI: geometric mean fluorescent intensity

Article Snippet: G-MDSCs were plated in 48-well plates or 24-well plates (Costar, Corning, NY) with RPMI 1640 medium supplemented with 10% fetal calf serum and then transfected with 50 nM Pvt1 siRNA or the negative control (Ribobio Co., Guangzhou, China) using Entranster-R (Engreen Biosystem Co., Beijing, China) according to the manufacturer’s instructions.

Techniques: Knockdown, In Vitro, Injection, Transfection, Negative Control, Quantitative RT-PCR, Activity Assay, Flow Cytometry

Journal: Molecular Cancer

Article Title: Long noncoding RNA Pvt1 regulates the immunosuppression activity of granulocytic myeloid-derived suppressor cells in tumor-bearing mice

doi: 10.1186/s12943-019-0978-2

Figure Lengend Snippet: c-myc is a potential downstream target of Pvt1 in G-MDSCs. a Scatter plot for protein-coding RNAs. b qRT-PCR was used to detect the mRNA level of c-myc in G-MDSCs sorted from spleen of WT mice and tumor tissues of TB mice. c , d After transfection with si-Pvt1, the mRNA and protein levels of c-myc in G-MDSCs isolated from tumor tissues were measured via qRT-PCR and western blot analyses. ** p < 0.01, and * p < 0.05

Article Snippet: G-MDSCs were plated in 48-well plates or 24-well plates (Costar, Corning, NY) with RPMI 1640 medium supplemented with 10% fetal calf serum and then transfected with 50 nM Pvt1 siRNA or the negative control (Ribobio Co., Guangzhou, China) using Entranster-R (Engreen Biosystem Co., Beijing, China) according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Transfection, Isolation, Western Blot

Journal: Molecular Cancer

Article Title: Long noncoding RNA Pvt1 regulates the immunosuppression activity of granulocytic myeloid-derived suppressor cells in tumor-bearing mice

doi: 10.1186/s12943-019-0978-2

Figure Lengend Snippet: Pvt1 knockdown reduces the ability of G-MDSCs to accelerate tumor progression and inhibit antitumor immune responses. Two groups of mice were given a s.c. injection of a mixture of LLCs and G-MDSCs transfected with si-Pvt1 (si-Pvt1 group) or si-NC (si-NC group). a Tumor volume was measured at the indicated time. b , c The proportions of CD8 + IFN-γ + CTLs and CD4 + IFN-γ + Th1 cells from draining lymph nodes, spleens and tumor tissues were analyzed via flow cytometry. ** p < 0.01, and * p < 0.05

Article Snippet: G-MDSCs were plated in 48-well plates or 24-well plates (Costar, Corning, NY) with RPMI 1640 medium supplemented with 10% fetal calf serum and then transfected with 50 nM Pvt1 siRNA or the negative control (Ribobio Co., Guangzhou, China) using Entranster-R (Engreen Biosystem Co., Beijing, China) according to the manufacturer’s instructions.

Techniques: Knockdown, Injection, Transfection, Flow Cytometry

Journal: Molecular Cancer

Article Title: Long noncoding RNA Pvt1 regulates the immunosuppression activity of granulocytic myeloid-derived suppressor cells in tumor-bearing mice

doi: 10.1186/s12943-019-0978-2

Figure Lengend Snippet: HIF-1α upregulates Pvt1 expression in G-MDSCs under hypoxic stress. a The mRNA level of HIF-1α in G-MDSCs sorted from spleens and tumor tissues of TB mice was detected using qRT-PCR. G-MDSCs isolated from spleens of TB mice were cultured in an incubator at 37 °C (20% O 2 , 5% CO 2 ) (normoxic conditions) or in a sealed box containing an anaerobic bag to consume oxygen (O 2 < 0.1%, 5% CO 2 ) (hypoxic conditions). b, c The mRNA and protein levels of HIF-1α were measured via qRT-PCR and western blot (WB) analyses, respectively. d Pvt1 expression was analyzed via qRT-PCR. YC-1, a specific inhibitor of HIF-1α, was used to block hypoxia. e, f HIF-1α and ( g ) Pvt1 expression in the normoxia, hypoxia, and hypoxia+YC-1 groups were detected via qRT-PCR and WB analyses. *** p < 0.001, ** p < 0.01, and * p < 0.05; ns: no significance

Article Snippet: G-MDSCs were plated in 48-well plates or 24-well plates (Costar, Corning, NY) with RPMI 1640 medium supplemented with 10% fetal calf serum and then transfected with 50 nM Pvt1 siRNA or the negative control (Ribobio Co., Guangzhou, China) using Entranster-R (Engreen Biosystem Co., Beijing, China) according to the manufacturer’s instructions.

Techniques: Expressing, Quantitative RT-PCR, Isolation, Cell Culture, Western Blot, Blocking Assay

Journal: Molecular Cancer

Article Title: LncRNA PVT1 up-regulation is a poor prognosticator and serves as a therapeutic target in esophageal adenocarcinoma

doi: 10.1186/s12943-019-1064-5

Figure Lengend Snippet: LncRNA PVT1 is amplified and overexpressed in EAC tumor tissues compared to Barrett’s epithelium and normal tissues. a. Analysis of PVT1 genomic alteration in TCGA dataset revealed PVT1 was amplified in over 20% of ESCA cases, and around 75% of ESCA cases containing both amplification and duplication (> 3 N) in total. b. Amplification of PVT1 gene in EAC is more common than that in ESCC. c. PVT1 expression levels significantly higher in amplification and duplication cases than that in LOH and diploid cases. d & e. Expressions PVT1 lncRNA and MYC mRNA were measured by qPCR and normalized to GAPDH in our own patient-cohort (156 cases)

Article Snippet: In addition, the expression of PVT1 as measured by qPCR in tumor tissues showed significant reduction when treated with PVT1 ASO compared with the control ASO group (Fig. ).

Techniques: Amplification, Expressing

Journal: Molecular Cancer

Article Title: LncRNA PVT1 up-regulation is a poor prognosticator and serves as a therapeutic target in esophageal adenocarcinoma

doi: 10.1186/s12943-019-1064-5

Figure Lengend Snippet: Correlation between expression of PVT1 and clinical characteristics. a. Higher expression of PVT1 lncRNA was significantly correlated with advanced tumor stage (T stage and N stage), as well as higher grade tumor and signet ring cell subtype. b. Kaplan-Meier analysis of OS time according to PVT1 and MYC expression level. EAC patients with higher PVT1 expression showed poorer survival than that of low expression (left). MYC expression is not associated with survival of patients with EAC. High expression was defined as ≥2 times of mean of normal tissue, whereas low expression was considered as < 2 times of mean of normal tissue. Log-rank probabilities between high and low expression were shown. Error bars, mean ± SEM. *, P < 0.05

Article Snippet: In addition, the expression of PVT1 as measured by qPCR in tumor tissues showed significant reduction when treated with PVT1 ASO compared with the control ASO group (Fig. ).

Techniques: Expressing

Journal: Molecular Cancer

Article Title: LncRNA PVT1 up-regulation is a poor prognosticator and serves as a therapeutic target in esophageal adenocarcinoma

doi: 10.1186/s12943-019-1064-5

Figure Lengend Snippet: PVT1 suppression by PVT1 ASOs inhibits EAC cells growth in vitro and in vivo. a. PVT1 expression in EAC cell lines (FLO1, SK-GT-4, JHESO, and OE19) and normal esophageal epithelial cell line HET-1A were determined by realtime qPCR analysis. b. PVT1 expression was analyzed by qPCR in PVT1 ASOs treated EAC cell lines. PVT1 specific antisense oligonucleotides (ASO4 and ASO5) reduced PVT1 expression in dose-dependent manner in three EAC cell lines. c and d. Inhibition of PVT1 by ASOs significantly suppressed colony formation ( c ) and decreased cell invasion ( d ) in both JHESO and OE19 cells in two individual PVT1 ASOs respectively. e . Average tumor volume in mice that treated with PVT1 ASO4 or control ASO via subcutaneous injection for 3 weeks (left). Actual tumor weights were retrieved from the mice at the termination of the experiment (middle). Representative tumors after scarified were shown (right). f. The expression of PVT1 in mouse tumor tissues measured by qPCR has been shown significant reduction upon treatment with PVT1 ASO4 compared to the control group. Error bars, mean ± SEM. *, P < 0.05; **, P < 0.01

Article Snippet: In addition, the expression of PVT1 as measured by qPCR in tumor tissues showed significant reduction when treated with PVT1 ASO compared with the control ASO group (Fig. ).

Techniques: In Vitro, In Vivo, Expressing, Inhibition, Control, Injection

Journal: Molecular Cancer

Article Title: LncRNA PVT1 up-regulation is a poor prognosticator and serves as a therapeutic target in esophageal adenocarcinoma

doi: 10.1186/s12943-019-1064-5

Figure Lengend Snippet: Suppression of PVT1 by ASOs reduces cancer stem cell (CSC) properties in EAC cells. a and b . Representative images (top) and the number (below) of tumorsphere formation showed that silencing of PVT1 by ASO4 and ASO5 suppressed sphere formation in both JHESO and OE19 cells. c . PVT1 ASO5 reduced the population of ALDH1+ cells (top: OE19; below: JHESO). JHESO and OE19 cells treated with PVT1 ASO5 at 10uM for 48 h and then labeling with ALDH1A1 using ALDH1 Labeling Kit. DEAB as negative control. Scale bar, 100 μm. Error bars, mean ± SEM. *, P < 0.05; **, P < 0.01

Article Snippet: In addition, the expression of PVT1 as measured by qPCR in tumor tissues showed significant reduction when treated with PVT1 ASO compared with the control ASO group (Fig. ).

Techniques: Labeling, Negative Control

Journal: Molecular Cancer

Article Title: LncRNA PVT1 up-regulation is a poor prognosticator and serves as a therapeutic target in esophageal adenocarcinoma

doi: 10.1186/s12943-019-1064-5

Figure Lengend Snippet: Mutual regulation of PVT1 and YAP1 in EAC cells. a . Reverse-phase protein arrays (RPPA) revealed changes in total YAP1 (left) and phosphorylated YAP1 (Ser 127) (right) treated with or without PVT1 ASOs. * p < 0.05. b . Correlation analysis (Pearson’s correlation) between PVT1 and YAP1 expression in 103 EAC patients. Pearson’s correlation (scatter plot) of expression levels between PVT1 and YAP1 in patients with EAC (r = 0.6319, p < 0.0001). c . Total YAP1 level and phosphor-YAP1 (Ser 127) or phosphor-LATS1(Ser 909) were determined by Western blot in OE19 and JHESO cell lines treated with PVT1 ASO at concentration indicated. d . Immunofluorescent staining of YAP1 and phosphor-LATS1 in OE19 and JHESO cell lines treated with PVT1 ASO. Scale bar; 25 μm. e . Level of YAP1 and its targets-SOX9 and CTGF as well as PVT1 level were determined by Western blot and qPCR in SKGT-4 cells with or without YAP1 induction (Doxycycline; DOX+)., f . YAP1 and PVT1 level was detected by q-PCR or western blot in YAP1 knockout clones (YAP1 KO1 and YAP1 KO2) with LentiCRISPR/Cas9 compared to control cells. PVT1 expression was dramatically decreased (bottom) upon knock out YAP1 in SKGT-4 EAC cells. Error bars, mean ± SEM. ** P < 0.01

Article Snippet: In addition, the expression of PVT1 as measured by qPCR in tumor tissues showed significant reduction when treated with PVT1 ASO compared with the control ASO group (Fig. ).

Techniques: Expressing, Western Blot, Concentration Assay, Staining, Knock-Out, Clone Assay, Control

Journal: Molecular Cancer

Article Title: LncRNA PVT1 up-regulation is a poor prognosticator and serves as a therapeutic target in esophageal adenocarcinoma

doi: 10.1186/s12943-019-1064-5

Figure Lengend Snippet: Simultaneous inhibition of PVT1 and YAP1 by their ASOs had better anti-tumor activity on EAC cells in vitro and in vivo. a. Expression of PVT1(left) and YAP1 (right) were detected in JHESO and OE19 cells treated with PVT1 ASOs, YAP1 ASOs or their combination by real-time q-PCR. b. Colony formation of JHESO (left) and OE19 (right) cells were significantly suppressed by PVT1 and YAP1 ASOs alone or in combination. c . Average tumor volume were demonstrated in mice that treated with PVT1 ASO, YAP1 ASO and their combination for 4 weeks. d . Tumors weights at the termination of the experiment were shown. e & f. Representative tumors ( e ) and mice weight ( f ) after 4 weeks were calculated as described as Materials&Methods. Error bars, mean ± SEM. E. *, P < 0.05; **, P < 0.01. g. Proposed model by which mutual regulation of PVT1 and YAP1

Article Snippet: In addition, the expression of PVT1 as measured by qPCR in tumor tissues showed significant reduction when treated with PVT1 ASO compared with the control ASO group (Fig. ).

Techniques: Inhibition, Activity Assay, In Vitro, In Vivo, Expressing

Journal: Molecular Cancer

Article Title: LncRNA PVT1 triggers Cyto-protective autophagy and promotes pancreatic ductal adenocarcinoma development via the miR-20a-5p/ULK1 Axis

doi: 10.1186/s12943-018-0845-6

Figure Lengend Snippet: ULK1 protein level parallels to that of PVT1 in a subset of human PDA tissues. a Western blot analysis of the expression of ULK1 protein in PDA tissues ( n = 20) and the corresponding adjacent non-tumor pancreatic specimens (n = 20). GAPDH acted as an endogenous control. T, tumor tissues; N, non-tumor pancreatic tissues; b The quantitation of the western blot results in (a). ULK1 protein expression was up-regulated in 13 PDA tissues compared with the corresponding adjacent non-tumor pancreatic specimens in 20 PDA tissues. The high value of ULK1 protein was defined as fold change > 1 ( n = 13); c The level of ULK1 mRNA in 20 PDA tissues was of no statistical significance compared to that of the corresponding adjacent non-tumor pancreatic specimens by qRT-PCR. Data are presented as the mean ± S.D. (n = 20); d PVT1 expression in PDA tissues from the 20 PDA cases based on qRT-PCR analysis. The high value of PVT1 was defined as fold change > 2 ( n = 15), the rest including down-regulation or no evident difference in expression in PDA tissues compared with PDA expression in the corresponding adjacent non-tumor tissue, was defined as low values ( n = 5); e PVT1 expression in PDA tissues based on ISH analysis and the percent of PVT1-positive samples in different groups; f The expression level of PVT1 mRNA was positively correlated with that of ULK1 protein in 20 PDA tissues. The statistical analysis was performed using Pearson’s correlation coefficient ( R = 0.8659, P < 0.0001); g The level of PVT1 mRNA in PDA cell lines by qRT-PCR. GAPDH acted as an endogenous control; h The expression level of PVT1 mRNA was positively correlated with that of ULK1 protein in PDA cell lines ( R = 0.8546, P = 0.0069). Data are presented as the mean ± S.D. from three independent experiments. * P < 0.05, ** P < 0.01 by Student’s t-test

Article Snippet: The human PVT1 3’UTR luciferase reporter construct (PVT1-wt) was generated by cloning the PVT1 mRNA 3’UTR sequence downstream of the psiCHECK luciferase reporter vector (Promega).

Techniques: Western Blot, Expressing, Quantitation Assay, Quantitative RT-PCR

Journal: Molecular Cancer

Article Title: LncRNA PVT1 triggers Cyto-protective autophagy and promotes pancreatic ductal adenocarcinoma development via the miR-20a-5p/ULK1 Axis

doi: 10.1186/s12943-018-0845-6

Figure Lengend Snippet: Regulation of PVT1 made contributions to autophagy in PDA cells. a PVT1 level was overexpressed by specific lentivirus transfection in Capan-2 and MIA PaCa-2 cells; b PVT1 level was markedly suppressed by specific sh-RNAs in SW1990 and HPAF-II cells; c The ultrastructure of cells with or without up-regulation of PVT1 in Capan-2 and MIA PaCa-2 cells was observed by transmission electron microscopy. Autophagic vacuoles were increased after overexpression of PVT1 in Capan-2 and MIA PaCa-2 cells. Scale bar, 2 μm; d The ultrastructure of cells with or without down-regulation of PVT1 in SW1990 and HPAF-II cells. Autophagic vacuoles were decreased after attenuation of PVT1 expression in SW1990 and HPAF-II cells. Scale bar, 1 μm. Data are presented as the mean ± S.D. ( n = 3). * P < 0.05, ** P < 0.01 and *** P < 0.001 by Student’s t-test

Article Snippet: The human PVT1 3’UTR luciferase reporter construct (PVT1-wt) was generated by cloning the PVT1 mRNA 3’UTR sequence downstream of the psiCHECK luciferase reporter vector (Promega).

Techniques: Transfection, Transmission Assay, Electron Microscopy, Over Expression, Expressing

Journal: Molecular Cancer

Article Title: LncRNA PVT1 triggers Cyto-protective autophagy and promotes pancreatic ductal adenocarcinoma development via the miR-20a-5p/ULK1 Axis

doi: 10.1186/s12943-018-0845-6

Figure Lengend Snippet: Detection of GFP-LC3 puncta with or without regulation of PVT1. a , b Representative images of GFP-LC3 puncta in PVT1-overexpressed cells (Capan-2 and MIA PaCa-2) ( a ) and PVT1-downregulated cells (SW1990 and HPAF-II) ( b ); c , d Quantitation of GFP-LC3 puncta in PVT1-overexpressed cells (Capan-2 and MIA PaCa-2) ( c ) and PVT1-downregulated cells (SW1990 and HPAF-II) ( d ). Enhanced expression remarkably increased the number of GFP-LC3 puncta. Meanwhile, down-regulation of PVT1 decreased the number of GFP-LC3 puncta. Data are presented as the mean ± S.D. ( n = 3). * P < 0.05 and ** P < 0.01 by Student’s t-test

Article Snippet: The human PVT1 3’UTR luciferase reporter construct (PVT1-wt) was generated by cloning the PVT1 mRNA 3’UTR sequence downstream of the psiCHECK luciferase reporter vector (Promega).

Techniques: Quantitation Assay, Expressing

Journal: Molecular Cancer

Article Title: LncRNA PVT1 triggers Cyto-protective autophagy and promotes pancreatic ductal adenocarcinoma development via the miR-20a-5p/ULK1 Axis

doi: 10.1186/s12943-018-0845-6

Figure Lengend Snippet: PVT1 played a vital role in cyto-protective autophagy and PDA cells growth in vitro. a MTS assay indicated that up-regulation of PVT1 enhanced cells growth in Capan-2 cells. Whereas, impairment of growth was observed after incubation with autophagy inhibitor (3-MA) (upper). Meanwhile, down-regulation of PVT1 suppressed cells growth in HPAF-II cells. However, restoration of growth occurred after treatment with autophagy inducer (rapamycin) (lower); b Colony formation assay implied that increased PVT1 expression promoted cells proliferation in Capan-2 cells. However, 3-MA diminished cells proliferation in PVT1-overexpresssed cells. On the other hand, decreased PVT1 expression inhibited cells proliferation in HPAF-II cells. But rescue of cells growth was observed after treatment with rapamycin; c Cell proliferation was investigated by EdU incorporation assays; d Cell cycle analysis indicated that the percentage of mitotic S phase cells was increased after overexpression of PVT1 in Capan-2 cells. Decreased percentage occurred after incubation with 3-MA. Decreased expression of PVT1 diminished S phase cells in HPAF-II cells. Rescue of S phase happened after induction of autophagy by rapamycin; e Apoptosis prohibited after up-regulation of PVT1 in Capan-2 cells. Inhibition of autophagy promoted apoptosis. Meanwhile, down-regulation of PVT1 contributed to apoptosis in HPAF-II cells. Autophagy induction by rapamycin diminished the effect of down-regulation of PVT1 to apoptosis. The data were represented as the means ± S.D. of three independent experiments in vitro . * P < 0.05 by Student’s t-test

Article Snippet: The human PVT1 3’UTR luciferase reporter construct (PVT1-wt) was generated by cloning the PVT1 mRNA 3’UTR sequence downstream of the psiCHECK luciferase reporter vector (Promega).

Techniques: In Vitro, MTS Assay, Incubation, Colony Assay, Expressing, Cell Cycle Assay, Over Expression, Inhibition

Journal: Molecular Cancer

Article Title: LncRNA PVT1 triggers Cyto-protective autophagy and promotes pancreatic ductal adenocarcinoma development via the miR-20a-5p/ULK1 Axis

doi: 10.1186/s12943-018-0845-6

Figure Lengend Snippet: PVT1 made an effort on autophagy via regulation of ULK1. a Western blot analysis of LC3b in Capan-2 and MIA PaCa-2 cells with or without up-regulation of PVT1. It was observed that the level of LC3b II was enhanced after increase of PVT1 expression; b LC3b II expression was decreased after down-regulation of PVT1; c Western blot analysis of LC3b in PVT1 stable-expressed Capan-2 and MIA PaCa-2 cells with or without depletion of ULK1. Suppression of LC3b II expression was detected after depletion of ULK1; d LC3b II expression was rescued after restoration of ULK1 in PVT1-knowndown SW1990 and HPAF-II cells

Article Snippet: The human PVT1 3’UTR luciferase reporter construct (PVT1-wt) was generated by cloning the PVT1 mRNA 3’UTR sequence downstream of the psiCHECK luciferase reporter vector (Promega).

Techniques: Western Blot, Expressing

Journal: Molecular Cancer

Article Title: LncRNA PVT1 triggers Cyto-protective autophagy and promotes pancreatic ductal adenocarcinoma development via the miR-20a-5p/ULK1 Axis

doi: 10.1186/s12943-018-0845-6

Figure Lengend Snippet: Knockdown of PVT1 prohibited PDA growth in vivo. a Down-regulation of PVT1 in HPAF-II cells inhibited tumor growth after 4 weeks in vivo ( n = 5 per group). Upper: negative control group. Lower: knockdown groups formed in nude mice subcutaneously injected with PVT1-silencing HPAF-II cells; b , c Growth curves ( b ) and tumor weight ( c ) of xenograft tumors after subcutaneous injection with PVT1-silencing HPAF-II cells. The tumor volumes were investigated every week after injection with PVT1-silencing HPAF-II cells ( n = 5). Diminished expression of PVT1 markedly attenuated the tumor growth in PDA; d , e Representative images ( d ) and photon influx ( e ) of xenograft tumors after subcutaneous injection with PVT1-silencing HPAF-II cells by IVIS ( n = 5 per group). f Representative images of HE staining, TUNEL, immunohistochemical staining of Ki67 and ULK1; g-i The quantitation of Ki67 ( g ), TUNEL ( h ) and ULK1 ( i ) was calculated. Immunohistochemical staining of Ki67 and ULK1 showed that decreased expression of PVT1 led to suppression of Ki67 proliferation index and ULK1 expression, while TUNEL positive cells were increased. The data were represented as the mean ± S.D. ( n = 5). * P < 0.05, ** P < 0.01 and *** P < 0.001 by Student’s t-test

Article Snippet: The human PVT1 3’UTR luciferase reporter construct (PVT1-wt) was generated by cloning the PVT1 mRNA 3’UTR sequence downstream of the psiCHECK luciferase reporter vector (Promega).

Techniques: In Vivo, Negative Control, Injection, Expressing, Staining, TUNEL Assay, Immunohistochemical staining, Quantitation Assay

Journal: Molecular Cancer

Article Title: LncRNA PVT1 triggers Cyto-protective autophagy and promotes pancreatic ductal adenocarcinoma development via the miR-20a-5p/ULK1 Axis

doi: 10.1186/s12943-018-0845-6

Figure Lengend Snippet: PVT1 modulates ULK1 expression by sponging miR-20a-5p. a The venn diagram of common microRNAs targeting PVT1 3’UTR and ULK1 3’UTR. Three microRNAs were identified, including miR-20a-5p, miR-302a-3p and miR-17-5p; b PVT1 biotin pull down assay was performed. The level of target microRNAs in the pull down of biotin-labelled PVT1 or negative control was investigated and quantified by qRT-PCR in HPAF-II cells; c Western blot analysis of ULK1 expression in HPAF-II cells with or without overexpression (upper) or suppression (lower) of target microRNAs (miR-20a-5p, miR-302a-3p, miR-17-5p). Interestingly, overexpression of miR-20a-5p suppressed ULK1 protein expression most. While inhibition of miR-20a-5p promoted ULK1 protein expression effectively; d RIP assay and qRT-PCR were conducted to detect the enrichment of PVT1 and miR-20a-5p by using AGO2 antibody in HPAF-II cells. Non-immune IgG acted as an internal control. U6 was used as a non-specific control. Enhanced enrichment of PVT1 and miR-20a-5p with AGO2 antibody was observed compared with IgG; e Bioinformatics prediction of miR-20a-5p binding sites in PVT1 3’UTR sequence using Starbase or ULK1 3’UTR sequence using TargetScan was presented; f Dual-luciferase assays showed diminished luciferase activity was observed when co-transfection of psiCHECK-ULK1-WT and miR-20a-5p occurred in HEK 293 T and HPAF-II cells; g Luciferase reporter assay implied that decreased luciferase activity occurred in HEK 293 T and HPAF-II cells co-transfected with psiCHECK-PVT1-WT and miR-20a-5p compared with luciferase reporter with mutant type of PVT1 and miR-20a-5p; h , i qRT-PCR analysis of miR-20a-5p with augment ( h ) of PVT1 expression in Capan-2 and MIA PaCa-2 cells or attenuation ( i ) of PVT1 expression in SW1990 and HPAF-II cells; j Overexpression of miR-20a-5p made no contribution to the level of PVT1 expression. Data are presented as the mean ± S.D. ( n = 3). * P < 0.05, ** P < 0.01 by Student’s t-test

Article Snippet: The human PVT1 3’UTR luciferase reporter construct (PVT1-wt) was generated by cloning the PVT1 mRNA 3’UTR sequence downstream of the psiCHECK luciferase reporter vector (Promega).

Techniques: Expressing, Pull Down Assay, Negative Control, Quantitative RT-PCR, Western Blot, Over Expression, Inhibition, Binding Assay, Sequencing, Luciferase, Activity Assay, Cotransfection, Reporter Assay, Transfection, Mutagenesis

Journal: Molecular Cancer

Article Title: LncRNA PVT1 triggers Cyto-protective autophagy and promotes pancreatic ductal adenocarcinoma development via the miR-20a-5p/ULK1 Axis

doi: 10.1186/s12943-018-0845-6

Figure Lengend Snippet: PVT1 is up-regulated in PDA, correlates with PDA progression and predicts prognosis. a , b Data mining of PVT1 in PDA patients gene profiling GSE16515 ( a ) and GSE15471 ( b ). PVT1 is overexpressed in PDA tissues compared with the corresponding adjacent non-tumor pancreatic specimens. Data are presented as the mean ± S.D.. ** P < 0.001 by Student’s t-test; c PVT1 expression in 68 PDA patients. Fold change > 2 is used as the cut-off value in the expression. The high value of PVT1 was defined as fold change > 2 ( n = 53), the rest was defined as low expression ( n = 15); d PVT1 expression is up-regulated in patients with stage III + IV ( n = 24) compared with patients with stage I + II ( n = 44). Data are presented as the mean ± S.D.. * P < 0.05 by Student’s t-test; e High expression of PVT1 predicts poor prognosis in 68 PDA patients. * P < 0.05 by the log-rank test

Article Snippet: The human PVT1 3’UTR luciferase reporter construct (PVT1-wt) was generated by cloning the PVT1 mRNA 3’UTR sequence downstream of the psiCHECK luciferase reporter vector (Promega).

Techniques: Expressing

Journal: Molecular Cancer

Article Title: LncRNA PVT1 triggers Cyto-protective autophagy and promotes pancreatic ductal adenocarcinoma development via the miR-20a-5p/ULK1 Axis

doi: 10.1186/s12943-018-0845-6

Figure Lengend Snippet: Clinicopathological characteristics of 68 pancreatic ductal adenocarcinoma patients

Article Snippet: The human PVT1 3’UTR luciferase reporter construct (PVT1-wt) was generated by cloning the PVT1 mRNA 3’UTR sequence downstream of the psiCHECK luciferase reporter vector (Promega).

Techniques: Expressing

Journal: Molecular Cancer

Article Title: LncRNA PVT1 triggers Cyto-protective autophagy and promotes pancreatic ductal adenocarcinoma development via the miR-20a-5p/ULK1 Axis

doi: 10.1186/s12943-018-0845-6

Figure Lengend Snippet: Schematic diagram shows the results of the study. PVT1 is up-regulated in PDA. Moreover, it triggers cyto-protective autophagy and promotes PDA cells growth and progression. It acts as a ceRNA to compete with miR-20a-5p, which leads to enhanced expression of ULK1, a key molecule involved in triggering autophagy initiation

Article Snippet: The human PVT1 3’UTR luciferase reporter construct (PVT1-wt) was generated by cloning the PVT1 mRNA 3’UTR sequence downstream of the psiCHECK luciferase reporter vector (Promega).

Techniques: Expressing