Journal: Cancer Cell International

Article Title: Silenced lncRNA DDX11-AS1 or up-regulated microRNA-34a-3p inhibits malignant phenotypes of hepatocellular carcinoma cells via suppression of TRAF5

doi: 10.1186/s12935-021-01847-6

Figure Lengend Snippet: Primer sequence

Article Snippet: The lentivirus containing sh-DDX11-AS1 and sh-NC, miR-34a-3p mimic and its NC were purchased from GenePharma Co. Ltd. (Shanghai, China). pcDDX11-AS1 was constructed by RiboBio Co., Ltd. (Guangdong, China).

Techniques:

Journal: Cancer Cell International

Article Title: Silenced lncRNA DDX11-AS1 or up-regulated microRNA-34a-3p inhibits malignant phenotypes of hepatocellular carcinoma cells via suppression of TRAF5

doi: 10.1186/s12935-021-01847-6

Figure Lengend Snippet: Elevated DDX11-AS1, TRAF5 and reduced miR-34a-3p are present in HCC tissues and cells. a DDX11-AS1, miR-34a-3p and TRAF5 expression in HCC tissues and adjacent tissues detected via RT-qPCR; b TRAF5 protein bands in HCC tissues and adjacent tissues; c TRAF5 protein expression in HCC tissues and adjacent tissues detected via western blot analysis; d The correlation between DDX11-AS1 and miR-34a-3p expression in HCC patients analyzed via Pearson test; e The correlation between DDX11-AS1 and TRAF5 expression in HCC patients analyzed via Pearson test; f The correlation between miR-34a-3p and TRAF5 expression in HCC patients analyzed via Pearson test; g DDX11-AS1, miR-34a-3p and TRAF5 expression in normal human hepatocytes LO2 and HCC cell lines SMMC-7721 and SK-hep1 detected via RT-qPCR; h TRAF5 protein bands in normal human hepatocytes LO2 and HCC cell lines SMMC-7721 and SK-hep1; i TRAF5 protein expression in normal human hepatocytes LO2 and HCC cell lines SMMC-7721 and SK-hep1 detected via western blot analysis; The measurement data were expressed as mean ± standard deviation. The t test was used for two-group comparison. One-way ANOVA was used for comparison among multiple groups, and Tukey’s post hoc test was used for pairwise comparison. Pearson test was used for correlation analysis. * vs LO2 cells, P < 0.05

Article Snippet: The lentivirus containing sh-DDX11-AS1 and sh-NC, miR-34a-3p mimic and its NC were purchased from GenePharma Co. Ltd. (Shanghai, China). pcDDX11-AS1 was constructed by RiboBio Co., Ltd. (Guangdong, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation

Journal: Cancer Cell International

Article Title: Silenced lncRNA DDX11-AS1 or up-regulated microRNA-34a-3p inhibits malignant phenotypes of hepatocellular carcinoma cells via suppression of TRAF5

doi: 10.1186/s12935-021-01847-6

Figure Lengend Snippet: Reduced DDX11-AS1 inhibits the proliferation, promotes apoptosis of HCC cells. a The cell growth curve detected by CCK-8 assay; b The cell proliferation rate detected via colony formation assay; c The apoptosis rate detected via Annexin V-FITC/PI double staining; The measurement data were expressed as mean ± standard deviation. The t test was used for two-group comparison. ^ vs the sh-NC group, P < 0.05

Article Snippet: The lentivirus containing sh-DDX11-AS1 and sh-NC, miR-34a-3p mimic and its NC were purchased from GenePharma Co. Ltd. (Shanghai, China). pcDDX11-AS1 was constructed by RiboBio Co., Ltd. (Guangdong, China).

Techniques: CCK-8 Assay, Colony Assay, Double Staining, Standard Deviation

Journal: Cancer Cell International

Article Title: Silenced lncRNA DDX11-AS1 or up-regulated microRNA-34a-3p inhibits malignant phenotypes of hepatocellular carcinoma cells via suppression of TRAF5

doi: 10.1186/s12935-021-01847-6

Figure Lengend Snippet: Reduced DDX11-AS1 inhibits the migration, invasion of HCC cells, promotes and represses the tumor growth in nude mice. a The cell migration detected by scratch test; b The cell invasion detected by Transwell assay; c Representative images of xenografted tumors in nude mice of each group; d Xenografted tumor volume in nude mice of each group; e Xenografted tumor weight in nude mice of each group; The measurement data were expressed as mean ± standard deviation. The t test was used for two-group comparison. ^ vs the sh-NC group, P < 0.05

Article Snippet: The lentivirus containing sh-DDX11-AS1 and sh-NC, miR-34a-3p mimic and its NC were purchased from GenePharma Co. Ltd. (Shanghai, China). pcDDX11-AS1 was constructed by RiboBio Co., Ltd. (Guangdong, China).

Techniques: Migration, Transwell Assay, Standard Deviation

Journal: Cancer Cell International

Article Title: Silenced lncRNA DDX11-AS1 or up-regulated microRNA-34a-3p inhibits malignant phenotypes of hepatocellular carcinoma cells via suppression of TRAF5

doi: 10.1186/s12935-021-01847-6

Figure Lengend Snippet: Reduced DDX11-AS1 increases miR-34a-3p to reduce TRAF5 expression. a DDX11-AS1, miR-34a-3p and TRAF5 expression detected by RT-qPCR; b TRAF5 protein bands; c TRAF5 protein expression detected by western blot analysis; d The binding sites of DDX11-AS1 and miR-34a-3p predicted via bioinformatics website https://cm.jefferson.edu/rna22/Precomputed/ ; e The regulatory relationship between DDX11-AS1 and miR-34a-3p verified via dual luciferase reporter gene assay; f The binding relationship between DDX11-AS1 and miR-34a-3p verified via RNA-pull down assay; g The targeting relationship between miR-34a-3p and TRAF5 predicted via bioinformatics website https://cm.jefferson.edu/rna22/Precomputed/ ; h The targeting relationship between miR-34a-3p and TRAF5 verified via luciferase activity assay; The measurement data were expressed as mean ± standard deviation. One-way ANOVA was used for comparison among multiple groups, and Tukey’s post hoc test was used for pairwise comparison after ANOVA analysis. ^ vs the sh-NC group, P < 0.05; # vs the mimic NC group, P < 0.05

Article Snippet: The lentivirus containing sh-DDX11-AS1 and sh-NC, miR-34a-3p mimic and its NC were purchased from GenePharma Co. Ltd. (Shanghai, China). pcDDX11-AS1 was constructed by RiboBio Co., Ltd. (Guangdong, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Binding Assay, Luciferase, Reporter Gene Assay, Pull Down Assay, Activity Assay, Standard Deviation

Journal: Cancer Cell International

Article Title: Silenced lncRNA DDX11-AS1 or up-regulated microRNA-34a-3p inhibits malignant phenotypes of hepatocellular carcinoma cells via suppression of TRAF5

doi: 10.1186/s12935-021-01847-6

Figure Lengend Snippet: DDX11-AS1 modulates the growth and metastasis of HCC cells through miR-34a-3p/TRAF5 axis. a , b DDX11-AS1, miR-34a-3p and TRAF5 expression after co-transfection detected by RT-qPCR; c TRAF5 protein expression after co-transfection detected by western blot analysis; d The cell growth curve detected by CCK-8 assay; e The cell proliferation rate detected via colony formation assay; f The apoptosis rate detected via Annexin V-FITC/PI double staining; The measurement data were expressed as mean ± standard deviation. t test was used for comparison between two groups, One-way ANOVA for comparison among multiple groups, and Tukey’s post hoc test for pairwise comparison. & vs the pcDDX11-AS1 + mimic NC group, P < 0.05

Article Snippet: The lentivirus containing sh-DDX11-AS1 and sh-NC, miR-34a-3p mimic and its NC were purchased from GenePharma Co. Ltd. (Shanghai, China). pcDDX11-AS1 was constructed by RiboBio Co., Ltd. (Guangdong, China).

Techniques: Expressing, Cotransfection, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Colony Assay, Double Staining, Standard Deviation

Journal: Cancer Cell International

Article Title: Silenced lncRNA DDX11-AS1 or up-regulated microRNA-34a-3p inhibits malignant phenotypes of hepatocellular carcinoma cells via suppression of TRAF5

doi: 10.1186/s12935-021-01847-6

Figure Lengend Snippet: DDX11-AS1 modulates the growth and metastasis of HCC cells through miR-34a-3p/TRAF5 axis. a , b DDX11-AS1, miR-34a-3p and TRAF5 expression after co-transfection detected by RT-qPCR; c TRAF5 protein expression after co-transfection detected by western blot analysis; d The cell growth curve detected by CCK-8 assay; e The cell proliferation rate detected via colony formation assay; f The apoptosis rate detected via Annexin V-FITC/PI double staining; The measurement data were expressed as mean ± standard deviation. The measurement data were expressed as mean ± standard deviation. t test was used for comparison between two groups, One-way ANOVA for comparison among multiple groups, and Tukey’s post hoc test for pairwise comparison. + vs the pcDDX11-AS1 + sh-NC group, P < 0.05

Article Snippet: The lentivirus containing sh-DDX11-AS1 and sh-NC, miR-34a-3p mimic and its NC were purchased from GenePharma Co. Ltd. (Shanghai, China). pcDDX11-AS1 was constructed by RiboBio Co., Ltd. (Guangdong, China).

Techniques: Expressing, Cotransfection, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Colony Assay, Double Staining, Standard Deviation

Journal: Cancer Cell International

Article Title: Silenced lncRNA DDX11-AS1 or up-regulated microRNA-34a-3p inhibits malignant phenotypes of hepatocellular carcinoma cells via suppression of TRAF5

doi: 10.1186/s12935-021-01847-6

Figure Lengend Snippet: DDX11-AS1 modulates the growth and metastasis of HCC cells through miR-34a-3p/TRAF5 axis. a The cell migration detected by scratch test; b The cell invasion detected by Transwell assay; c Representative images of xenografted tumors in nude mice; d Xenografted tumor volume in nude mice of each group; e Xenografted tumor weight in nude mice of each group; The measurement data were expressed as mean ± standard deviation. The measurement data were expressed as mean ± standard deviation. t test was used for comparison between two groups, One-way ANOVA for comparison among multiple groups, and Tukey’s post hoc test for pairwise comparison. & vs the pcDDX11-AS1 + mimic NC group, P < 0.05

Article Snippet: The lentivirus containing sh-DDX11-AS1 and sh-NC, miR-34a-3p mimic and its NC were purchased from GenePharma Co. Ltd. (Shanghai, China). pcDDX11-AS1 was constructed by RiboBio Co., Ltd. (Guangdong, China).

Techniques: Migration, Transwell Assay, Standard Deviation

Journal: Cancer Cell International

Article Title: Silenced lncRNA DDX11-AS1 or up-regulated microRNA-34a-3p inhibits malignant phenotypes of hepatocellular carcinoma cells via suppression of TRAF5

doi: 10.1186/s12935-021-01847-6

Figure Lengend Snippet: DDX11-AS1 modulates the growth and metastasis of HCC cells through miR-34a-3p/TRAF5 axis. a The cell migration detected by scratch test; b The cell invasion detected by Transwell assay; c Representative images of xenografted tumors in nude mice of each group; d Xenografted tumor volume in nude mice of each group; e Xenografted tumor weight in nude mice of each group; The measurement data were expressed as mean ± standard deviation. The measurement data were expressed as mean ± standard deviation. t test was used for comparison between two groups, One-way ANOVA for comparison among multiple groups, and Tukey’s post hoc test for pairwise comparison. + vs the pcDDX11-AS1 + sh-NC group, P < 0.05

Article Snippet: The lentivirus containing sh-DDX11-AS1 and sh-NC, miR-34a-3p mimic and its NC were purchased from GenePharma Co. Ltd. (Shanghai, China). pcDDX11-AS1 was constructed by RiboBio Co., Ltd. (Guangdong, China).

Techniques: Migration, Transwell Assay, Standard Deviation

Journal: Bioscience Reports

Article Title: A regulatory circuitry comprising TP53, miR-29 family, and SETDB1 in non-small cell lung cancer

doi: 10.1042/BSR20180678

Figure Lengend Snippet: ( A ) Dual luciferase assay of 293T cells co-transfected with the luciferase constructs containing the SETDB1 3′-UTR_WT or SETDB1 3′-UTR_Mut as well as miR-29s mimics or Scr NC. ( B ) SETDB1 protein levels was assessed in PC14, A549, and BEAS-2B cells. ( C ) SETDB1 mRNA levels and ( D ) SETDB1 protein levels after transfection of miR-29s mimics or Scr NC into A549 and PC14 cells. ( E ) SETDB1 mRNA levels and ( F ) SETDB1 protein levels after transfection of miR-29s inhibitors or Scr NC into BEAS-2B cells. * P <0.05; ** P <0.01; *** P <0.001

Article Snippet: The siRNAs specifically targetting SETDB1 (siSETDB1) and TP53 (siTP53) were obtained commercially (GenePharma, Shanghai, China).

Techniques: Luciferase, Transfection, Construct

Journal: Bioscience Reports

Article Title: A regulatory circuitry comprising TP53, miR-29 family, and SETDB1 in non-small cell lung cancer

doi: 10.1042/BSR20180678

Figure Lengend Snippet: miR-29s mRNA expression after transfection of pEGFP-TP53 vector or pEGFP-N1 control vector into ( A ) A549 cells and ( B ) PC14 cells. ( C ) SETDB1 protein level and ( D ) SETDB1 mRNA expression after transfection of pEGFP-TP53 vector or pEGFP-N1 control vector into A549 and PC14 cells. ( E ) miR-29s mRNA expression, ( F ) SETDB1 mRNA expression and ( G ) SETDB1 protein level after transfection of siTP53 or Scr NC into BEAS-2B cells. ** P <0.01; *** P <0.001

Article Snippet: The siRNAs specifically targetting SETDB1 (siSETDB1) and TP53 (siTP53) were obtained commercially (GenePharma, Shanghai, China).

Techniques: Expressing, Transfection, Plasmid Preparation, Control

Journal: Bioscience Reports

Article Title: A regulatory circuitry comprising TP53, miR-29 family, and SETDB1 in non-small cell lung cancer

doi: 10.1042/BSR20180678

Figure Lengend Snippet: ( A ) TP53 mRNA levels and ( B ) TP53 protein levels ( C , D ) miR-29s levels after transfection of siSETDB1 scrambled oligonucleotides as the negative control into A549 and PC14 cells. ( E ) TP53 mRNA levels and ( F ) TP53 protein levels and ( G ) miR-29s levels after transfection of pcDNA3.1-SETDB1 vector or pcDNA3.1 control vector into BEAS-2B cells. ** P <0.01; *** P <0.001

Article Snippet: The siRNAs specifically targetting SETDB1 (siSETDB1) and TP53 (siTP53) were obtained commercially (GenePharma, Shanghai, China).

Techniques: Transfection, Negative Control, Plasmid Preparation, Control

Journal: Bioscience Reports

Article Title: A regulatory circuitry comprising TP53, miR-29 family, and SETDB1 in non-small cell lung cancer

doi: 10.1042/BSR20180678

Figure Lengend Snippet: ( A ) SETDB1 mRNA levels and ( B ) TP53 mRNA levels in primary NSCLC samples and their paired adjacent normal samples. ( C ) SETDB1 and TP53 mRNA levels were inversely correlated in primary NSCLC samples. ( D – F ) Pearson correlation analysis between miR-29s and SETDB1 mRNA levels, as well as between miR-29s and TP53 mRNA levels in primary NSCLC samples. *** P <0.001

Article Snippet: The siRNAs specifically targetting SETDB1 (siSETDB1) and TP53 (siTP53) were obtained commercially (GenePharma, Shanghai, China).

Techniques:

Journal: Bioscience Reports

Article Title: A regulatory circuitry comprising TP53, miR-29 family, and SETDB1 in non-small cell lung cancer

doi: 10.1042/BSR20180678

Figure Lengend Snippet: ( A ) TP53 and Suv39h1 mRNA levels after transfection of miR-29s mimics or Scr NC into A549 and PC14 cells. ( B ) TP53 and Suv39h1 mRNA levels after transfection of miR-29s inhibitors or Scr NC into BEAS-2B cells. Histone H3K9 methylation status in ( C ) NSCLC cells (A549 and PC14) with overexpressed miR-29s and ( D ) BEAS-2B cells with downexpressed miR-29s were examined by Western blot using antibodies specific for H3K9me2 and H3K9me3 (di- and trimethylation of H3K9, respectively); expression levels of histone H3 protein were used as an internal control. ( E ) Schematic illustration of the proposed TP53/ miR-29s /SETDB1 regulatory circuitry in NSCLC. ** P <0.01; *** P <0.001

Article Snippet: The siRNAs specifically targetting SETDB1 (siSETDB1) and TP53 (siTP53) were obtained commercially (GenePharma, Shanghai, China).

Techniques: Transfection, Methylation, Western Blot, Expressing, Control