Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long noncoding RNA MALAT1 potentiates growth and inhibits senescence by antagonizing ABI3BP in gallbladder cancer cells

doi: 10.1186/s13046-019-1237-5

Figure Lengend Snippet: Primer sequences for reverse transcription quantitative polymerase chain reaction

Article Snippet: The ABI3BP-vector plasmid (50 ng/mL) and vector-NC plasmid (50 ng/mL) purchased from GenePharma Co., Ltd. (Shanghai, China) were respectively transfected into cells.

Techniques:

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long noncoding RNA MALAT1 potentiates growth and inhibits senescence by antagonizing ABI3BP in gallbladder cancer cells

doi: 10.1186/s13046-019-1237-5

Figure Lengend Snippet: ABI3BP is poorly expressed in GBC tissues and cells. a , microarray data analysis of GSE74048 dataset. The abscissa indicates the sample type and the ordinate indicates the differential genes. Histogram on the upper right represents color gradation and rectangle refers to the expression level. b , the correlation analysis between expression of MALAT1 and ABI3BP in GBC tissues; c , the relative expression of ABI3BP in normal gallbladder tissues (n = 16) and GBC tissues (n = 48) detected by immunohistochemistry (400 ×). d , the relative expression of ABI3BP in the GBC cells (GBC-SD, SGC-996, NOZ and OCUG1) and HGBECs. * p < 0.05 vs. the normal gallbladder tissue or HGBEC cell line. The data was measurement data and expressed as mean value ± standard deviation. Comparison was analyzed by independent sample t -test between two groups and by one-way analysis of variance among multiple groups, followed by Turkey’s post-hoc test. The experiment was repeated 3 times. ABI3BP, ABI family member 3 binding protein; GBC, gallbladder cancer; HGBECs, human gallbladder epithelial cells

Article Snippet: The ABI3BP-vector plasmid (50 ng/mL) and vector-NC plasmid (50 ng/mL) purchased from GenePharma Co., Ltd. (Shanghai, China) were respectively transfected into cells.

Techniques: Microarray, Expressing, Immunohistochemistry, Standard Deviation, Binding Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long noncoding RNA MALAT1 potentiates growth and inhibits senescence by antagonizing ABI3BP in gallbladder cancer cells

doi: 10.1186/s13046-019-1237-5

Figure Lengend Snippet: ABI3BP over-expression restrains the proliferation and metastasis of GBC cells while contributes to senescence. a and b , the proliferation of GBC-SD cells detected by EdU assay (200 ×). c and d , the protein expression of Ki67 and PCNA relative to β-actin in GBC-SD cells determined by Western blot analysis. e , the migration of GBC-SD cells detected by scratch test. f , the invasion of GBC-SD cells detected by Transwell assay (200 ×). g , the senescence of GBC-SD cells detected by SA-β-gal staining (400 ×). * p < 0.05 vs. the GBC cells manipulated with vector-NC. The data was measurement data and expressed as mean value ± standard deviation. Comparison was analyzed by independent sample t -test. The experiment was repeated 3 times. ABI3BP, ABI family member 3 binding protein; GBC, gallbladder cancer; EdU, 5-ethynyl-2′-deoxyuridine; PCNA, proliferating cell nuclear antigen; SA-β-gal, senescence-associated β-galactosidase; NC, negative control

Article Snippet: The ABI3BP-vector plasmid (50 ng/mL) and vector-NC plasmid (50 ng/mL) purchased from GenePharma Co., Ltd. (Shanghai, China) were respectively transfected into cells.

Techniques: Over Expression, EdU Assay, Expressing, Western Blot, Migration, Transwell Assay, Staining, Plasmid Preparation, Standard Deviation, Binding Assay, Negative Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long noncoding RNA MALAT1 potentiates growth and inhibits senescence by antagonizing ABI3BP in gallbladder cancer cells

doi: 10.1186/s13046-019-1237-5

Figure Lengend Snippet: MALAT1 down-regulates the expression of ABI3BP through EZH2 recruitment in the promoter region of ABI3BP. a , the subcellular localization of MALAT1 predicted on lncATLAS website. b , Blast comparison between sequences of MALAT1 and ABI3BP promoter. c , the subcellular localization of MALAT1 detected by FISH assay (400 ×). d , the enrichment of EZH2 by MALAT1 detected by RIP assay. e , the relative expression of EZH2 relative to β-actin detected by RNA pull down. f , EZH2 enrichment in the ABI3BP promoter region detected by ChIP assay. g , the relative expression of ABI3BP determined by RT-qPCR. * p < 0.05 vs. the cells treated with sh-NC, Bio-probe-NC or DMSO. The data was measurement data and expressed as mean value ± standard deviation. Comparison was analyzed by independent sample t -test between two groups and by one-way analysis of variance among multiple groups, followed by Turkey’s post-hoc test. The experiment was repeated 3 times. MALAT1, metastasis associated lung adenocarcinoma transcript 1; ABI3BP, ABI family member 3 binding protein; EZH2, enhancer of zeste homolog 2; FISH, fluorescence in situ hybridization; RIP, RNA immunoprecipitation; ChIP, chromatin immunoprecipitation; RT-qPCR, reverse transcription quantitative polymerase chain reaction; NC, negative control; DMSO, dimethyl sulfoxide

Article Snippet: The ABI3BP-vector plasmid (50 ng/mL) and vector-NC plasmid (50 ng/mL) purchased from GenePharma Co., Ltd. (Shanghai, China) were respectively transfected into cells.

Techniques: Expressing, Quantitative RT-PCR, Standard Deviation, Binding Assay, Fluorescence, In Situ Hybridization, Immunoprecipitation, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Negative Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long noncoding RNA MALAT1 potentiates growth and inhibits senescence by antagonizing ABI3BP in gallbladder cancer cells

doi: 10.1186/s13046-019-1237-5

Figure Lengend Snippet: EZH2 down-regulates the expression of ABI3BP via H3K27 methylation. a , the relative expression of H3K27me3 determined by Western blot analysis following the treatment of GSK343 (EZH2 inhibitor) or sh-MALAT1. b , the relative expression of H3K27me3 determined by Western blot analysis following the treatment of UTX (histone demethylase). c , the relative expression of ABI3BP determined by Western blot analysis following the treatment of GSK343 (EZH2 inhibitor) or sh-MALAT1. d , the relative expression of ABI3BP determined by Western blot analysis following the treatment of UTX (histone demethylase). * p < 0.05 vs. the cells treated with sh-NC or DMSO. # p < 0.05 vs. the cells transfected with sh-MALAT1. The data was measurement data and expressed as mean value ± standard deviation. Comparison was analyzed by independent sample t -test between two groups and by one-way analysis of variance among multiple groups, followed by Turkey’s post-hoc test. The experiment was repeated 3 times. EZH2, enhancer of zeste homolog 2; ABI3BP, ABI family member 3 binding protein; H3K27me3, trimethylation of lysine 27 on histone H3 protein subunit; MALAT1, metastasis associated lung adenocarcinoma transcript 1; UTX, ubiquitously transcribed tetratricopeptide repeat on chromosome X; NC, negative control; DMSO, dimethyl sulfoxide

Article Snippet: The ABI3BP-vector plasmid (50 ng/mL) and vector-NC plasmid (50 ng/mL) purchased from GenePharma Co., Ltd. (Shanghai, China) were respectively transfected into cells.

Techniques: Expressing, Methylation, Western Blot, Transfection, Standard Deviation, Binding Assay, Negative Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long noncoding RNA MALAT1 potentiates growth and inhibits senescence by antagonizing ABI3BP in gallbladder cancer cells

doi: 10.1186/s13046-019-1237-5

Figure Lengend Snippet: Over-expressed ABI3BP and silenced MALAT1 and EZH2 impede GBC cell proliferation, migration, invasion and tumorigenic potential while accelerate senescence. a , representative images of resected tumors from nude mice and tumor weights. b , the relative expression of ABI3BP determined by immunohistochemistry (400 ×). c , the relative expression of H3K27me3 relative to β-actin determined by Western blot analysis. d , the relative expression of Ki67 and PCNA relative to β-actin determined by Western blot analysis. e , the cell migration detected by scratch test. f , the cell invasion detected by Transwell assay (200 ×). g , the cell senescence detected by SA-β-gal staining (400 ×). * p < 0.05 vs. nude mice treated with vector-NC or sh-NC. The data was measurement data and expressed as mean value ± standard deviation. Comparison was analyzed by one-way analysis of variance among multiple groups, followed by Turkey’s post-hoc test. The experiment was repeated 3 times. ABI3BP, ABI family member 3 binding protein; MALAT1, metastasis associated lung adenocarcinoma transcript 1; EZH2, enhancer of zeste homolog 2; GBC, gallbladder cancer; H3K27me3, trimethylation of lysine 27 on histone H3 protein subunit; PCNA, proliferating cell nuclear antigen; SA-β-gal, senescence-associated β-galactosidase; NC, negative control

Article Snippet: The ABI3BP-vector plasmid (50 ng/mL) and vector-NC plasmid (50 ng/mL) purchased from GenePharma Co., Ltd. (Shanghai, China) were respectively transfected into cells.

Techniques: Migration, Expressing, Immunohistochemistry, Western Blot, Transwell Assay, Staining, Plasmid Preparation, Standard Deviation, Binding Assay, Negative Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long noncoding RNA MALAT1 potentiates growth and inhibits senescence by antagonizing ABI3BP in gallbladder cancer cells

doi: 10.1186/s13046-019-1237-5

Figure Lengend Snippet: Suppression of proliferation, invasion, migration and tumorigenic potential induced by MALAT1 silencing could be rescued by ABI3BP silencing. a , the proliferation determined by EdU assay (200 ×). b , the cell migration detected by scratch test. c , the cell invasion detected by Transwell assay (200 ×). d , representative images of resected tumors from nude mice and tumor weights. e , the relative expression of ABI3BP determined by immunohistochemistry (400 ×). * p < 0.05 vs. cells or nude mice treated with sh-MALAT1. The data was measurement data and expressed as mean value ± standard deviation. Comparison was analyzed by independent sample t -test between two groups. The experiment was repeated 3 times

Article Snippet: The ABI3BP-vector plasmid (50 ng/mL) and vector-NC plasmid (50 ng/mL) purchased from GenePharma Co., Ltd. (Shanghai, China) were respectively transfected into cells.

Techniques: Migration, EdU Assay, Transwell Assay, Expressing, Immunohistochemistry, Standard Deviation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long noncoding RNA MALAT1 potentiates growth and inhibits senescence by antagonizing ABI3BP in gallbladder cancer cells

doi: 10.1186/s13046-019-1237-5

Figure Lengend Snippet: The mechanism of MALAT1 on GBC. MALAT1 silencing or ABI3BP restoration can facilitate the treatment of patients with GBC by exerting suppressive effects on GBC cell growth properties both in vitro and in vivo accompanied by strengthened the ability of cell senescence

Article Snippet: The ABI3BP-vector plasmid (50 ng/mL) and vector-NC plasmid (50 ng/mL) purchased from GenePharma Co., Ltd. (Shanghai, China) were respectively transfected into cells.

Techniques: In Vitro, In Vivo

Journal: Cancer research

Article Title: KDM5B is essential for the hyper-activation of PI3K/AKT signaling in prostate tumorigenesis

doi: 10.1158/0008-5472.CAN-20-0505

Figure Lengend Snippet: (A) Left panel, western blotting analysis showing the protein levels of KDM5B, pAKT, P110α, and P85 in LNCaP KDM5B-KO and the parental control human PCa cells. Right panel, quantification analysis of pAKT, P110α, and P85 in LNCaP KDM5B-KO and the control cells. (B) A comparison of the cell proliferation between LNCaP KDM5B-KO and the control cells. (C) Immunofluorescence (IF) images and analysis of PIP3 in LNCaP KDM5B-KO cells. Left panel, IF images showing the levels and membrane localization of PIP3 in LNCaP KDM5B-KO cells and the control cells. Right panel, quantification of the fluorescence intensities of PIP3 in LNCaP KDM5B-KO and the control cells. Error bars represent means ± SD (20 cells/group). (D) RNA-Seq peaks showing the changes of KDM5B, IRS1, PIK3CA, and PIK3R1 expression between LNCaP KDM5B-KO and the control cells. (E) Quantitative RT-PCR analysis to show the relative mRNA levels of IRS1, PIK3CA, and PIK3R1 in LNCaP KDM5B-KO cells. (F) Top panel, western blotting analysis showing the protein levels of KDM5B, P110α and P85 in LNCaP KDM5B-KO and the control cells at indicated time points in CHX chase experiments. Bottom panel, quantification of protein remaining for P110α and P85 at indicated time points in KDM5B-KO and the control cells.

Article Snippet: For PIK3CA promoter reporter assay, cells were transfected with 200 ng empty pLightSwitch_Prom reporter vector, 50 ng, 100 ng or 200 ng of PIK3CA -promoter-Luc reporter plasmids (Switchgear Genomics, S706543).

Techniques: Western Blot, Immunofluorescence, Fluorescence, RNA Sequencing Assay, Expressing, Quantitative RT-PCR

Journal: Cancer research

Article Title: KDM5B is essential for the hyper-activation of PI3K/AKT signaling in prostate tumorigenesis

doi: 10.1158/0008-5472.CAN-20-0505

Figure Lengend Snippet: (A) A schematic showing the positions of 5 amplicons relative to the PIK3CA transcription start site (TSS). (B) ChIP analysis of KDM5B levels at the indicated regions near PIK3CA TSS in LNCaP KDM5B-KO and the parental control cells. (C) ChIP analysis of H3K4me3 levels at the indicated regions near PIK3CA TSS in LNCaP KDM5B-KO and the control cells. (D) Top panel, a schematic showing that KDM5B regulates the transcription of PIK3CA through affecting promoter activities. Bottom panel, comparisons of luciferase activities of the PIK3CA promoter between LNCaP KDM5B-KO cells and the parental control cells. (E) Luciferase activities showing the effects of KDM5B restoration on the PIK3CA promoter activities in LNCaP KDM5B-KO cells.

Article Snippet: For PIK3CA promoter reporter assay, cells were transfected with 200 ng empty pLightSwitch_Prom reporter vector, 50 ng, 100 ng or 200 ng of PIK3CA -promoter-Luc reporter plasmids (Switchgear Genomics, S706543).

Techniques: Luciferase