Journal: International Journal of Nanomedicine

Article Title: Docetaxel-loaded solid lipid nanoparticles suppress breast cancer cells growth with reduced myelosuppression toxicity

doi: 10.2147/IJN.S70919

Figure Lengend Snippet: Genes regulated by Taxotere ® and docetaxel-loaded solid lipid nanoparticles were confirmed by quantitative polymerase chain reaction and immunoblotting. Notes: Cell cycle-related genes of E2F8 ( A ) and OIP5 ( B ), proliferation-related genes of NASP ( C ) and SOD2 ( D ), and apoptosis-related genes of PDCD4 ( E ) and PIK3R2 ( F ) were chosen for detection by quantitative polymerase chain reaction and immunoblotting. In quantitative polymerase chain reaction detection, mock-treated cells were set as the control and samples were normalized with the control. In immunoblotting detection, β-actin was used as the loading control. Abbreviations: BSN, blank solid lipid nanoparticle; DSN, docetaxel-loaded solid lipid nanoparticle; GLU, glucose; qPCR, quantitative polymerase chain reaction; TAX, Taxotere.

Article Snippet: Primary antibodies of rabbit anti-β-actin, E2f8, MRE11A, ERBB3, IGFBP6, ATF3, CCNG2, SOD2, IGFBP3, CADM1, PDCD4, GADD45A, and MKI67 were purchased from Beijing Biosynthesis Technology Co., Ltd., (Beijing, People’s Republic of China), rabbit anti-MCM6, OIP5, and NASP were purchased from Proteintech Group, Inc., (Chicago, IL, USA), rabbit anti-FAM172A and MYB were purchased from Abgent, Inc. (San Diego, CA, USA) and rabbit anti-ATRX was purchased from GeneTex, Inc. (Irvine, CA, USA).

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: International Journal of Nanomedicine

Article Title: Docetaxel-loaded solid lipid nanoparticles suppress breast cancer cells growth with reduced myelosuppression toxicity

doi: 10.2147/IJN.S70919

Figure Lengend Snippet: Primers used for quantitative polymerase chain reaction

Article Snippet: Primary antibodies of rabbit anti-β-actin, E2f8, MRE11A, ERBB3, IGFBP6, ATF3, CCNG2, SOD2, IGFBP3, CADM1, PDCD4, GADD45A, and MKI67 were purchased from Beijing Biosynthesis Technology Co., Ltd., (Beijing, People’s Republic of China), rabbit anti-MCM6, OIP5, and NASP were purchased from Proteintech Group, Inc., (Chicago, IL, USA), rabbit anti-FAM172A and MYB were purchased from Abgent, Inc. (San Diego, CA, USA) and rabbit anti-ATRX was purchased from GeneTex, Inc. (Irvine, CA, USA).

Techniques: Sequencing

Journal: Molecular Oncology

Article Title: The Sp1/FOXC1/HOTTIP/LATS2/YAP/β‐catenin cascade promotes malignant and metastatic progression of osteosarcoma

doi: 10.1002/1878-0261.12760

Figure Lengend Snippet: FOXC1 played an essential role sustaining multiple malignant phenotypes of OS cells, both in vitro and in vivo . (A) Upon transfecting SW1353 (left panel) and 143B (right panel) cells with shFOXC1‐1, shFOXC1‐2, or control shRNA (shNC), the expression of FOXC1 protein was determined by western blot. The viability, long‐term proliferation, migration, and invasion of indicated cells at indicated time points were determined by MTT assay (B), colony‐forming assay (C), wound‐healing assay (D), and Transwell invasion assay (E), respectively, and compared between shNC and shFOXC1 cells. (F–H) shNC, shFOXC1‐1, or shFOXC1‐2 SW1353 and 143B cells were injected subcutaneously to establish xenograft tumors. The pictures of isolated xenografts after 28 days were shown in F, the growth curve in G, and the weights in H. (I–K) shNC, shFOXC1‐1, or shFOXC1‐2 SW1353 and 143B cells were injected intravenously to establish lung metastasis. The pictures of isolated lung tissues after 36 days were shown in I and microscopic metastasis detected upon HE staining shown in J and K. Error bars represent standard deviation. Student's t ‐test with four biological independent replicates was used to determine statistical significance; * P < 0.05, ** P < 0.01, *** P < 0.001, when compared to shNC cells. Scale bars: 1000 μm (D), 250 μm (E), and 100 μm (J).

Article Snippet: For HOTTIP shRNA (shHOTTIP), we designed four shRNA sequences shHOTTIP #1: 5′‐AAA AGC ATC TCA AAT TAA GCT TTG CCT CGA GGC AAA GCT TAA TTT GAG ATG C‐3′; shHOTTIP #2: 5′‐AAA AGG GAC TGA ATT CTT CGA GAT TTC TCG AGA AAT CTC AAG AAT TCA GTC CC‐3′; shHOTTIP #3: 5′‐AAA AGG TGC ACC TTA TTG ATC AAA TCT CGA GAT TTG AAT AAG GTG CAC C‐3′; shHOTTIP #4: 5′‐AAA AGC AGC CAA CAA ACT GAC TTG CCT CGA GGC AAG TCA GTT TGT TGG CTG C‐3′), cloned them into OIP5 shRNA lentiviral plasmid (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and generated lentiviral particles.

Techniques: In Vitro, In Vivo, Control, shRNA, Expressing, Western Blot, Migration, MTT Assay, Wound Healing Assay, Transwell Invasion Assay, Injection, Isolation, Staining, Standard Deviation

Journal: Molecular Oncology

Article Title: The Sp1/FOXC1/HOTTIP/LATS2/YAP/β‐catenin cascade promotes malignant and metastatic progression of osteosarcoma

doi: 10.1002/1878-0261.12760

Figure Lengend Snippet: HOTTIP recruited EZH1 and LSD1 to LATS2 promoter, enhanced promoter methylation, and suppressed the expression of LATS2. (A) The percentage distribution of HOTTIP in the nucleus and cytoplasm was examined by qRT‐PCR. GAPDH and U1 were detected as the marker for cytoplasm and nucleus, respectively. (B) EZH2 and LSD1 were identified as the potential interacting partner of HOTTIP using rpiseq software. (C) The interaction between HOTTIP and EZH2 or LSD1 was examined by RIP assay and presented as fold enrichment values relative to the amount of HOTTIP bound to IgG. (D, E) The expression of LATS2 in shHOTTIP‐1 and shHOTTIP‐2 vs. shNC cells was determined on the mRNA level by qRT‐PCR (D) and western blot (E), respectively. The levels of β‐catenin, YAP1, and p‐YAP1 were also detected by western blot (E). (F–H) SW1353 or 143B cells were transfected with siRNA targeting EZH2 (si‐EZH2‐1 and si‐EZH2‐2), LSD1 (si‐LSD1‐ and si‐LSD1‐2), or control (si‐NC). The expression levels of EZH2 (F), LSD1 (G), and LATS2 (H) were detected by qRT‐PCR. (I) The occupancy of EZH2, H3K27me3, LSD1, and H3K4me2 on LATS2 promoter in indicated cells was determined by ChIP analysis and presented as a fold value relative to the amount of IgG bound to the promoter. Error bars represent standard deviation. Student's t ‐test with three biological independent replicates was used to determine statistical significance; * P < 0.05, ** P < 0.01.

Article Snippet: For HOTTIP shRNA (shHOTTIP), we designed four shRNA sequences shHOTTIP #1: 5′‐AAA AGC ATC TCA AAT TAA GCT TTG CCT CGA GGC AAA GCT TAA TTT GAG ATG C‐3′; shHOTTIP #2: 5′‐AAA AGG GAC TGA ATT CTT CGA GAT TTC TCG AGA AAT CTC AAG AAT TCA GTC CC‐3′; shHOTTIP #3: 5′‐AAA AGG TGC ACC TTA TTG ATC AAA TCT CGA GAT TTG AAT AAG GTG CAC C‐3′; shHOTTIP #4: 5′‐AAA AGC AGC CAA CAA ACT GAC TTG CCT CGA GGC AAG TCA GTT TGT TGG CTG C‐3′), cloned them into OIP5 shRNA lentiviral plasmid (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and generated lentiviral particles.

Techniques: Methylation, Expressing, Quantitative RT-PCR, Marker, Software, Western Blot, Transfection, Control, Standard Deviation

Journal: Cancer control : journal of the Moffitt Cancer Center

Article Title: Opa-Interacting Protein 5 Expression in Human Glioma Tissues Is Essential to the Biological Function of U251 Human Malignant Glioma Cells.

doi: 10.1177/1073274820968914

Figure Lengend Snippet: Figure 1. Representative immunohistochemical staining of OIP5 expression in normal brain and glioma tissues. Immunohistochemical staining of OIP5 protein using an anti-OIP5 antibody in (A) normal human brain tissue (200), (B) grade I glioma (200), (C) grade II glioma (200), (D) grade III glioma (200), and (E) grade IV glioma (200).

Article Snippet: Sections were then incubated at 4 C overnight with a primary antibody to OIP5 (mouse antihuman, dilution 1:50, Catalog No. TA810617, OriGene, Rockville, MD, USA), and negative controls were incubated with PBS alone.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Cancer control : journal of the Moffitt Cancer Center

Article Title: Opa-Interacting Protein 5 Expression in Human Glioma Tissues Is Essential to the Biological Function of U251 Human Malignant Glioma Cells.

doi: 10.1177/1073274820968914

Figure Lengend Snippet: Figure 2. OIP5 expression in glioma cell lines and OIP5–siRNA inhibition of OIP5 expression in cells. (A): RT-PCR revealed that the OIP5 gene is abundantly or moderately expressed in U251, U373, A172, and glioblastoma of unknown origin cell lines. (B) and (C): Effect of OIP5 knockdown on the levels of OIP5 protein in 293 T cells as detected by western blot analysis. D-G: Analysis of GFP expression in U251 cells after infection with a lentiviral vector expressing GFP. (D) and (E): U251 cells in the control group (200). (F) and (G): GFP expression in the OIP5-siRNA group (200), reflecting infection efficiency in U251 cells. (H) Infection with Lentivirus-siOIP5 inhibited OIP5 mRNA expression in U251 cells as determined by RT-PCR. (**P < 0.05).

Article Snippet: Sections were then incubated at 4 C overnight with a primary antibody to OIP5 (mouse antihuman, dilution 1:50, Catalog No. TA810617, OriGene, Rockville, MD, USA), and negative controls were incubated with PBS alone.

Techniques: Expressing, Inhibition, Reverse Transcription Polymerase Chain Reaction, Knockdown, Western Blot, Infection, Plasmid Preparation, Control

Journal: Cancer control : journal of the Moffitt Cancer Center

Article Title: Opa-Interacting Protein 5 Expression in Human Glioma Tissues Is Essential to the Biological Function of U251 Human Malignant Glioma Cells.

doi: 10.1177/1073274820968914

Figure Lengend Snippet: Figure 3. Celigo Image Cytometer analysis shows that knocking down OIP5 significantly inhibits U251 cell proliferation. (A) and (B) The number of cells in each colony was counted over 5 days. (C) and (D) Cell number in the OIP5-siRNA group significantly decreased from days 3 to 5 compared to the control group (P < 0.05).

Article Snippet: Sections were then incubated at 4 C overnight with a primary antibody to OIP5 (mouse antihuman, dilution 1:50, Catalog No. TA810617, OriGene, Rockville, MD, USA), and negative controls were incubated with PBS alone.

Techniques: Cytometry, Control

Journal: Cancer control : journal of the Moffitt Cancer Center

Article Title: Opa-Interacting Protein 5 Expression in Human Glioma Tissues Is Essential to the Biological Function of U251 Human Malignant Glioma Cells.

doi: 10.1177/1073274820968914

Figure Lengend Snippet: Figure 4. MTT assay showed that OIP5 knockdown inhibits U251 cell proliferation. Infection with lentivirus-siOIP5 significantly inhibited cell proliferation in U251 cells from day 3 to day 5. Cell counts were determined by MTT assay over 5 days (P < 0.05).

Article Snippet: Sections were then incubated at 4 C overnight with a primary antibody to OIP5 (mouse antihuman, dilution 1:50, Catalog No. TA810617, OriGene, Rockville, MD, USA), and negative controls were incubated with PBS alone.

Techniques: MTT Assay, Knockdown, Infection

Journal: Cancer control : journal of the Moffitt Cancer Center

Article Title: Opa-Interacting Protein 5 Expression in Human Glioma Tissues Is Essential to the Biological Function of U251 Human Malignant Glioma Cells.

doi: 10.1177/1073274820968914

Figure Lengend Snippet: Figure 6. Knockdown of OIP5 promotes apoptosis in U251 cells. FCM shows that knocking out OIP5 expression increases U251 cell apoptosis compared with the control group (**P < 0.05).

Article Snippet: Sections were then incubated at 4 C overnight with a primary antibody to OIP5 (mouse antihuman, dilution 1:50, Catalog No. TA810617, OriGene, Rockville, MD, USA), and negative controls were incubated with PBS alone.

Techniques: Knockdown, Expressing, Control

Journal: Cancer control : journal of the Moffitt Cancer Center

Article Title: Opa-Interacting Protein 5 Expression in Human Glioma Tissues Is Essential to the Biological Function of U251 Human Malignant Glioma Cells.

doi: 10.1177/1073274820968914

Figure Lengend Snippet: Figure 5. Knockdown of OIP5 induces cell cycle arrest in U251 cells. The effect of silencing of OIP5 on the cell cycle was evaluated by flow cytometric assay. A significant increase in the cell population in the G1 and G2 phase and a significant decrease of the cell population in the S phase were detected in the OIP5-siRNA group compared with control U251 cells (*P < 0.05, **P < 0.01).

Article Snippet: Sections were then incubated at 4 C overnight with a primary antibody to OIP5 (mouse antihuman, dilution 1:50, Catalog No. TA810617, OriGene, Rockville, MD, USA), and negative controls were incubated with PBS alone.

Techniques: Knockdown, Flow Cytometry, Control

Journal: Cancers

Article Title: Prognostic and Therapeutic Potential of the OIP5 Network in Papillary Renal Cell Carcinoma

doi: 10.3390/cancers13174483

Figure Lengend Snippet: Upregulation of OIP5 associates with adverse features of pRCC and predicts poor overall survival. ( A ) IHC staining for OIP5 was performed using a RCC TMA; typical images of OIP5 staining in the adjacent kidney (AJK) and pRCC tumor tissues are presented. ( B ) Quantification of OIP5 IHC staining by H-score in the indicated tissues; means ± standard deviations (SDs) are graphed. Statistical analyses were performed using 2-tailed Student’s t -test; ***: p < 0.001 compared to the respective AJK tissues, $$$: p < 0.001 compared to T1 tumors. ( C – F ) OIP5 mRNA expressions in the indicated setting were analyzed using the TCGA dataset organized by UALCAN . Student’s t -test ( C ) and other indicated paired statistics were provided by UALCAN. *: p < 0.05, **: p < 0.01, ***: p < 0.001 compared to normal kidney tissues; $: p < 0.05, $$: p < 0.01 compared to T1P ( D ), Stage 2 ( E ), and N0 ( F ); ##: p < 0.01, ###: p < 0.001 compared to T2P ( D ) and Stage 3 ( E ). ( G ) Survival analysis was performed using the TCGA Pancancer pRCC dataset within cBioPortal. Logrank test was performed. Cutoff point used to separate the high- and low-OIP5 expression groups was ≥2 z-score or 2SD. The graph was produced using tools provided by cBioPortal. The median months overall survival for patients in the high-OIP5 group was 15.48 months.

Article Snippet: OIP5 cDNA plasmid was obtained from Origene (Cat: RG202255, Rockville, MD, USA) and subcloned into pBABE-puro retroviral plasmid (From Dr. Tak Mak at University of Toronto).

Techniques: Immunohistochemistry, Staining, Expressing, Produced

Journal: Cancers

Article Title: Prognostic and Therapeutic Potential of the OIP5 Network in Papillary Renal Cell Carcinoma

doi: 10.3390/cancers13174483

Figure Lengend Snippet: OIP5 promotes oncogenic processes of ACHN cells in vitro and in vivo. ( A ) ACHN empty vector (EV) and OIP5 stable lines. Western blot was carried out using anti-OIP5 and Actin antibodies. OIP5 expression was normalized to Actin and presented at fold changes to OIP5 expression in EV cells. ( B ) ACHN EV and ACHN OIP5 cells were seeded in 6-well plate at 10 5 cell/well; cell numbers were recorded at the indicated days. Experiments were repeated three times; means ± SDs are graphed. Statistical analysis was performed using 2-way ANOVA. ***: p < 0.001 between the two curves. ( C ) The indicated cells were seeded at the indicated number in 6-well plates. Colonies were formed following 2 weeks culture. Experiments were repeated three times; means ± SDs are graphed. **: p < 0.01 compared to the respective EV by Student’s t -test (2-way). ( D , E ) Invasion and soft agar assays were repeated 3 times; means ± SDs are graphed. **: p < 0.01, ***: p < 0.001 compared to the respective EV control by Student’s t -test (2-way). ( F , G ) Xenografts were produced in NOS/SCID mice (5 mice per group) using ACHN EV cells and ACHN OIP5 cells. Means ± SEM (standard error of the mean) are graphed; ***: p < 0.001 between the two curved by two-way ANOVA ( F ). Kaplan-Meier curve; statistical analysis was performed using logrank test ( G ).

Article Snippet: OIP5 cDNA plasmid was obtained from Origene (Cat: RG202255, Rockville, MD, USA) and subcloned into pBABE-puro retroviral plasmid (From Dr. Tak Mak at University of Toronto).

Techniques: In Vitro, In Vivo, Plasmid Preparation, Western Blot, Expressing, Control, Produced

Journal: Cancers

Article Title: Prognostic and Therapeutic Potential of the OIP5 Network in Papillary Renal Cell Carcinoma

doi: 10.3390/cancers13174483

Figure Lengend Snippet: OIP5 induces network alterations during pRCC tumorigenesis. ( A ) GSEA (gene set enrichment analysis) on differentially expressed genes derived from the comparison of ACHN OIP5 tumors to ACHN EV tumors was performed with FGSEA within the Galaxy platform. The MSigDB oncogenic gene sets (C6) collection was used. ( B ) Summary of the major oncogenic gene sets affected in ACHN OIP5 tumors (see for individual gene sets affected). ( C ) Enrichment of the indicated gene set within the MSigDB hallmark gene sets collection (see for individual gene sets affected).

Article Snippet: OIP5 cDNA plasmid was obtained from Origene (Cat: RG202255, Rockville, MD, USA) and subcloned into pBABE-puro retroviral plasmid (From Dr. Tak Mak at University of Toronto).

Techniques: Derivative Assay, Comparison

Journal: Cancers

Article Title: Prognostic and Therapeutic Potential of the OIP5 Network in Papillary Renal Cell Carcinoma

doi: 10.3390/cancers13174483

Figure Lengend Snippet: Pathway enrichment of OIP5 DEGs. DEGs were first defined as p .adj < 0.05 and fold changes > |1.5| in the comparison of ACHN OIP5 tumors ( n = 3) vs. ACHN EV tumors ( n = 3). ( A ) Pathway enrichment in these DEGs was then performed using the Metascape platform. ( B ) Clustering of DEGs in ACHN OIP5 tumors and ACHN EV tumors. ( C ) The number of overlapping genes between primary (patient) pRCC-derived DEGs and DEGs obtained from xenografts at fold change > |1.5|. ( D , E ) The indicated DEGs were analyzed for expression in the histological subtypes of pRCC using the UALCAN platform . DEGs positively ( D ) and negatively ( E ) predict shortening of OS (see for details). **: p < 0.01, ***: p < 0.001 in comparison to normal kidney tissues.

Article Snippet: OIP5 cDNA plasmid was obtained from Origene (Cat: RG202255, Rockville, MD, USA) and subcloned into pBABE-puro retroviral plasmid (From Dr. Tak Mak at University of Toronto).

Techniques: Comparison, Derivative Assay, Expressing

Journal: Cancers

Article Title: Prognostic and Therapeutic Potential of the OIP5 Network in Papillary Renal Cell Carcinoma

doi: 10.3390/cancers13174483

Figure Lengend Snippet: Characterization of Overlap66 genes.

Article Snippet: OIP5 cDNA plasmid was obtained from Origene (Cat: RG202255, Rockville, MD, USA) and subcloned into pBABE-puro retroviral plasmid (From Dr. Tak Mak at University of Toronto).

Techniques:

Journal: Cancers

Article Title: Prognostic and Therapeutic Potential of the OIP5 Network in Papillary Renal Cell Carcinoma

doi: 10.3390/cancers13174483

Figure Lengend Snippet: PLK1 inhibitor reduces OIP5-mediated pRCC tumorigenesis. ( A , B ) RNA-seq and real-time PCR analyses of PLK1 expression in ACHN EV and ACHN OIP5 tumors. RNA-seq was performed in 3 each from ACHN EV and ACHN OIP5 tumors. ( C ) ACHN EV and ACHN OIP5 cells were treated with DMSO (−) or the PLK1 inhibitor (BI2536) in 40 nM PLK1 inhibitor for 72 h, followed by quantification of cell cycle distributions. Experiments were repeated 3 times; means ± SEMs are graphed. **: p < 0.01, ***: p < 0.001, ****: p < 0.0001 in the indicated comparisons by 2-tailed Student’s test. ( D – G ) Mice bearing ACHN EV or ACHN OIP5 tumors were treated with vehicle or BI2536 (50 mg/kg) intravenously. The overall profiles of tumor growth in the vehicle treated setting ( D ); tumor volumes were recorded following treatments ( E – G ). Statistical analyses were performed using 2-tailed Student’s t -test; *: p < 0.05, ***: p < 0.001. ( H ) Kaplan Meier curve for the indicated mice reaching endpoints. Statistical analysis was performed using logrank test.

Article Snippet: OIP5 cDNA plasmid was obtained from Origene (Cat: RG202255, Rockville, MD, USA) and subcloned into pBABE-puro retroviral plasmid (From Dr. Tak Mak at University of Toronto).

Techniques: RNA Sequencing, Real-time Polymerase Chain Reaction, Expressing

Journal: PeerJ

Article Title: LncRNA OIP5-AS1 modulates the proliferation and apoptosis of Jurkat cells by sponging miR-181c-5p to regulate IL-7 expression in myasthenia gravis

doi: 10.7717/peerj.13454

Figure Lengend Snippet: Primer sequences.

Article Snippet: Several components were procured for this assay, including short hairpin RNAs (shRNA) targeting OIP5-AS1 (shOIP5-AS1) and its negative control (shRNA-NC), miR-181c-5p mimics and its negative control, and miR-181c-5p inhibitor and its negative control were designed and synthesized by GenScript (Nanjing, China).

Techniques: