【摘要】 目的 探讨PKCι在维持KRAS突变结肠癌生存中的作用及相关分子机制。方法 选取携KRAS突变的HCT116细胞和不携KRAS突变的RKO细胞为研究材料，两种细胞分别设置空白对照组(Control组)、ATM抑制组、shRNA-prkci组及shRNA-kras组。流式细胞术检测结肠癌细胞的凋亡及增殖情况。免疫印迹法(Western blot)和实时荧光定量PCR(qRT-PCR)检测KRAS、YAP1、PKCι的相对表达量。免疫荧光实验检测YAP1的表达。结果 在携KRAS突变的HCT116细胞中，PKCι的抑制伴随着细胞凋亡和生长抑制(均P<0.05);并且抑制PKCι可显著下调HCT116中YAP1的表达。而在不携带KRAS突变的RKO细胞中这种效应不明显。结论 PKCι通过调控YAP1维持KRAS突变结肠癌生存。PKCι与YAP1为潜在的结肠癌治疗靶标。更多还原

【Abstract】 Objective To explore the effect of PKCι on maintaining the survival process of KRAS mutation colon cancer and its molecular mechanism.Methods HCT116 and RKO, colon cancer cells were selected and divided into blank control group, ATM group, shRNA-prkci group and shRNA-kras group. The apoptosis was determined by flow cytometry. Protein level and mRNA level were detected by western blot and qPCR. Cell location and relative expression of YAP1 were validated by immunofluorescence. Results In HCT116 cells carrying KRAS gene mutations, PKCι inhibition was accompanied by apoptosis and growth inhibition(P<0.05). Furthermore, inhibition of PKCι significantly down-regulated YAP1 expression in HCT116. This effect was not obvious in RKO cells without KAS mutation. Conclusion PKCι maintains the survival of KRAS mutant colon cancer by regulating the expression of YAP1.PKCι and YAP1 are potential targets for colon cancer with KRAS mutation.更多还原