Journal: Oncogenesis
Article Title: Androgen receptor variant 7 exacerbates hepatocarcinogenesis in a c-MYC-driven mouse HCC model
doi: 10.1038/s41389-023-00449-3
Figure Lengend Snippet: A RNA-seq transcriptome analysis showed that 150 genes that were upregulated by c-MYC were repressed by AR-FL and/or AR-V7 co-expression; and 31 genes that were downregulated by c-MYC were upregulated by AR-FL and/or AR-V7 co-expression in both male (left) and female (right) mice at 20 dpi respectively. Each blue line represents differential expression levels (fold change in reference to control, Y axis) in the tumors of the respective combination of injected gene(s), X axis. Red lines highlight the differential expression levels of Cldn7 in respective samples. B Quantitative RT-PCR for expressions of Cldn7 in respective groups at 20 dpi. Expression of Cldn7 in male mice of c-MYC/p53KO (hep-c-MYC/p53KO) tumor at 11 dpi (pink bar), indicating that Cldn7 was repressed in p53-deficient conditions and hence was likely regulated by p53 [70–72]. C Expression of p53 in respective mouse groups in male and female, indicating p53 was upregulated by c-MYC but not significantly affected by AR-FL or AR-V7 co-expression. D Immunofluorescent images of c-MYC (red), Cldn7 FLAG (green), and DNA (blue) in the liver of hep-c-MYC/p53KO/Cldn7 FLAG mouse at 2 dpi, showing co-expression of the respective transgenes in the same cells. Scale bar = 50 µm E , Macroscopic phenotypes of the livers in male mice at 11 dpi injected with the expression vectors of c-MYC (c-MYC) ( n = 5), c-MYC under p53-deficient condition (c-MYC/p53KO) ( n = 6), or c-MYC and Cldn7 FLAG under p53-deficient condition (c-MYC/p53KO/Cldn7 FLAG ) ( n = 5). F Liver to body weight ratio of the samples presented in E , indicating that the exacerbation of c-MYC-driven HCC under p53-deficient conditions was completely reversed by co-expression of Cldn7 FLAG . Statistical significance by one-way ANOVA with Tukey’s multiple comparisons test for B , C , and F ; * p < 0.05; ** p < 0.001; *** p < 0.0001. Error bars indicate mean ± SEM.
Article Snippet: The pT3-based c-MYC expression vector pT3-c-MYC (#92046) [ ], the SB100 transposase expression vector pCMV-SB100 (#34879) [ ], the CRISPR/Cas9-base mouse p53 knockout vector pX330-p53 (#59910) [ ], and the full-length human androgen receptor vector pLENTI6.3/AR-GC-E2325 (#85128) were obtained from the Addgene Repository (Watertown, MA).
Techniques: RNA Sequencing, Expressing, Quantitative Proteomics, Control, Injection, Quantitative RT-PCR
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![A RNA-seq transcriptome analysis showed that 150 genes that were upregulated by c-MYC were repressed by AR-FL and/or AR-V7 co-expression; and 31 genes that were downregulated by c-MYC were upregulated by AR-FL and/or AR-V7 co-expression in both male (left) and female (right) mice at 20 dpi respectively. Each blue line represents differential expression levels (fold change in reference to control, Y axis) in the tumors of the respective combination of injected gene(s), X axis. Red lines highlight the differential expression levels of Cldn7 in respective samples. B Quantitative RT-PCR for expressions of Cldn7 in respective groups at 20 dpi. Expression of Cldn7 in male mice of c-MYC/p53KO (hep-c-MYC/p53KO) tumor at 11 dpi (pink bar), indicating that Cldn7 was repressed in <t>p53-deficient</t> conditions and hence was likely regulated by p53 [70–72]. C Expression of p53 in respective mouse groups in male and female, indicating p53 was upregulated by c-MYC but not significantly affected by AR-FL or AR-V7 co-expression. D Immunofluorescent images of c-MYC (red), Cldn7 FLAG (green), and DNA (blue) in the liver of hep-c-MYC/p53KO/Cldn7 FLAG mouse at 2 dpi, showing co-expression of the respective transgenes in the same cells. Scale bar = 50 µm E , Macroscopic phenotypes of the livers in male mice at 11 dpi injected with the expression vectors of c-MYC (c-MYC) ( n = 5), c-MYC under p53-deficient condition (c-MYC/p53KO) ( n = 6), or c-MYC and Cldn7 FLAG under p53-deficient condition (c-MYC/p53KO/Cldn7 FLAG ) ( n = 5). F Liver to body weight ratio of the samples presented in E , indicating that the exacerbation of c-MYC-driven HCC under p53-deficient conditions was completely reversed by co-expression of Cldn7 FLAG . Statistical significance by one-way ANOVA with Tukey’s multiple comparisons test for B , C , and F ; * p < 0.05; ** p < 0.001; *** p < 0.0001. Error bars indicate mean ± SEM.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_2460/pmc09902460/pmc09902460__41389_2023_449_Fig4_HTML.jpg)
