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Fig. 2 | Chinese Journal of Cancer

Fig. 2

From: Synthetic lethal short hairpin RNA screening reveals that ring finger protein 183 confers resistance to trametinib in colorectal cancer cells

Fig. 2

RNF183 expression was increased after trametinib treatment, and its knockdown synergistically repressed the proliferation of CRC cells in combination with trametinib. RNF183 mRNA abundance was examined using quantitative real-time PCR (qPCR), and its protein level was detected using Western blotting in HT29 (a) and HCT116 cells (b) with or without 20 nmol/L trametinib for 48 h. Arrows indicate the band of RNF183, which was obviously induced by trametinib. Effects of trametinib treatment in combination with RNF183 overexpression or knockdown on the proliferation of HT29 (c) and HCT116 cells (d) measured using MTT assay. RNF183 conferred resistance to trametinib-induced inhibition on proliferation in both cell lines. Effects of trametinib treatment in combination with RNF183 overexpression or knockdown on the apoptosis of HT29 (e) and HCT116 cells (f). Effects of trametinib treatment in combination with RNF183 overexpression or knockdown on lactate dehydrogenase (LDH) release of HT29 (g) and HCT116 cells (h). Experiments were repeated three times. Data are presented as mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ***P < 0.001. Tra trametinib, Ctrl stable cell lines transfected with pcDNA4-myc/his, RNF183 stable cell lines transfected with pcDNA4-myc/his-RNF183, shNC stable cell lines transfected with lentiviral vectors of negative control shRNA, shRNF183 stable cell lines transfected with lentiviral vectors of RNF183-targeted shRNA

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