human wt g401 rhabdoid cell line  (ATCC)

Journal: Cancer Management and Research

Article Title: Long Non-Coding RNA XIST Promotes Wilms Tumor Progression Through the miR-194-5p/YAP Axis

doi: 10.2147/CMAR.S297842

Figure Lengend Snippet: miR-194-5p can bind to both XIST and YAP in WT tissues. XIST lncRNA regulates WT progression through the miR-194-5p/YAP axis. ( A ) XIST 3ʹ-UTR wild-type (XIST-wt) sequence containing the miR-194-5p binding site and sequence of the mutant (XIST-mut) miR-194-5p binding site. ( B and C ) Luciferase reporter gene assay (images and histograms) showed lower luciferase activity for miR-194-5p and XIST-wt than XIST-mut ( P < 0.05). TRAF6 was used as an internal control to verify the integrity of the luciferase gene reporter assay. ( D ) YAP 3ʹ-UTR wild-type (YAP-wt) sequence containing the miR-194-5p binding site and sequence of the mutant (YAP-mut) miR-194-5p binding site. ( E and F ) Luciferase reporter gene assay (images and histograms) showed lower luciferase activity for miR-194-5p and YAP-wt than YAP-mut ( P < 0.05). TRAF6 was used as an internal control to verify the integrity of the luciferase gene reporter assay. ( G ) XIST lncRNA expression and ( H ) miR-194-5p in WT G401 cells and normal renal epithelial HK2 cells. ( I ) RT-qPCR analysis of miR-194-5p after transfection of lentiviral XIST, NC, and sh-XIST in WT G401 cells. ( J ) Western blot analysis showed that YAP protein expression can be regulated by miR-194-5p and XIST. One-way ANOVA or two-tailed t -test was performed for comparisons between the two groups. * P < 0.05, *** P < 0.001.

Article Snippet: The packaged lentiviral vectors were then transfected into the human WT G401 rhabdoid cell line (ATCC, USA) according to the manufacturer’s protocol and were collected for analysis after 48 h. The miR-194-5p mimic and NC mimic plasmids (GenePharma, Shanghai, China) were transfected into the human WT G401 cell line using Lipofectamine TM RNAiMAX (Invitrogen, USA) according to the manufacturer’s protocol.

Techniques: Sequencing, Binding Assay, Mutagenesis, Luciferase, Reporter Gene Assay, Activity Assay, Control, Reporter Assay, Expressing, Quantitative RT-PCR, Transfection, Western Blot, Two Tailed Test

Journal: Cancer Management and Research

Article Title: Long Non-Coding RNA XIST Promotes Wilms Tumor Progression Through the miR-194-5p/YAP Axis

doi: 10.2147/CMAR.S297842

Figure Lengend Snippet: XIST promotes the proliferation, migration, and invasion of G401 cells, and inhibits apoptosis in vitro. ( A and B ) Stable XIST overexpression (lentiviral XIST and negative control, NC) and XIST knockdown (lentiviral sh-RNA and negative control, sh-NC) were successfully established in WT G401 cells. ( C ) CCK-8 cell viability assay profiles showed that XIST overexpression enhanced G401 cell proliferation, while ( D ) XIST knockdown decreased cell proliferation. ( E ) Flow cytometry showed that XIST overexpression decreased apoptosis while XIST knockdown promoted apoptosis. ( F ) Scratch assay (100×magnification) ( G ) and transwell assay showed that XIST overexpression promoted G401 cell migration and invasion in vitro (100×magnification). Three independent replicates were performed. One-way ANOVA or two-tailed t -test was performed for comparisons between the two groups. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The packaged lentiviral vectors were then transfected into the human WT G401 rhabdoid cell line (ATCC, USA) according to the manufacturer’s protocol and were collected for analysis after 48 h. The miR-194-5p mimic and NC mimic plasmids (GenePharma, Shanghai, China) were transfected into the human WT G401 cell line using Lipofectamine TM RNAiMAX (Invitrogen, USA) according to the manufacturer’s protocol.

Techniques: Migration, In Vitro, Over Expression, Negative Control, Knockdown, CCK-8 Assay, Viability Assay, Flow Cytometry, Wound Healing Assay, Transwell Assay, Two Tailed Test

Journal: Cancer Management and Research

Article Title: Long Non-Coding RNA XIST Promotes Wilms Tumor Progression Through the miR-194-5p/YAP Axis

doi: 10.2147/CMAR.S297842

Figure Lengend Snippet: miR-194-5p can bind to both XIST and YAP in WT tissues. XIST lncRNA regulates WT progression through the miR-194-5p/YAP axis. ( A ) XIST 3ʹ-UTR wild-type (XIST-wt) sequence containing the miR-194-5p binding site and sequence of the mutant (XIST-mut) miR-194-5p binding site. ( B and C ) Luciferase reporter gene assay (images and histograms) showed lower luciferase activity for miR-194-5p and XIST-wt than XIST-mut ( P < 0.05). TRAF6 was used as an internal control to verify the integrity of the luciferase gene reporter assay. ( D ) YAP 3ʹ-UTR wild-type (YAP-wt) sequence containing the miR-194-5p binding site and sequence of the mutant (YAP-mut) miR-194-5p binding site. ( E and F ) Luciferase reporter gene assay (images and histograms) showed lower luciferase activity for miR-194-5p and YAP-wt than YAP-mut ( P < 0.05). TRAF6 was used as an internal control to verify the integrity of the luciferase gene reporter assay. ( G ) XIST lncRNA expression and ( H ) miR-194-5p in WT G401 cells and normal renal epithelial HK2 cells. ( I ) RT-qPCR analysis of miR-194-5p after transfection of lentiviral XIST, NC, and sh-XIST in WT G401 cells. ( J ) Western blot analysis showed that YAP protein expression can be regulated by miR-194-5p and XIST. One-way ANOVA or two-tailed t -test was performed for comparisons between the two groups. * P < 0.05, *** P < 0.001.

Article Snippet: Human WT G401 and normal renal tubular epithelial HK-2 cell lines (ATCC, USA) were cultured in DMEM medium containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (Gibco, USA) at 37°C, in a 5% CO 2 humidified incubator.

Techniques: Sequencing, Binding Assay, Mutagenesis, Luciferase, Reporter Gene Assay, Activity Assay, Control, Reporter Assay, Expressing, Quantitative RT-PCR, Transfection, Western Blot, Two Tailed Test

Journal: Cancer Management and Research

Article Title: Long Non-Coding RNA XIST Promotes Wilms Tumor Progression Through the miR-194-5p/YAP Axis

doi: 10.2147/CMAR.S297842

Figure Lengend Snippet: XIST promotes the proliferation, migration, and invasion of G401 cells, and inhibits apoptosis in vitro. ( A and B ) Stable XIST overexpression (lentiviral XIST and negative control, NC) and XIST knockdown (lentiviral sh-RNA and negative control, sh-NC) were successfully established in WT G401 cells. ( C ) CCK-8 cell viability assay profiles showed that XIST overexpression enhanced G401 cell proliferation, while ( D ) XIST knockdown decreased cell proliferation. ( E ) Flow cytometry showed that XIST overexpression decreased apoptosis while XIST knockdown promoted apoptosis. ( F ) Scratch assay (100×magnification) ( G ) and transwell assay showed that XIST overexpression promoted G401 cell migration and invasion in vitro (100×magnification). Three independent replicates were performed. One-way ANOVA or two-tailed t -test was performed for comparisons between the two groups. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Human WT G401 and normal renal tubular epithelial HK-2 cell lines (ATCC, USA) were cultured in DMEM medium containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (Gibco, USA) at 37°C, in a 5% CO 2 humidified incubator.

Techniques: Migration, In Vitro, Over Expression, Negative Control, Knockdown, CCK-8 Assay, Viability Assay, Flow Cytometry, Wound Healing Assay, Transwell Assay, Two Tailed Test