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目的:探究胶原三股螺旋重复蛋白1(CTHRC1)对卵巢癌细胞顺铂耐药的作用及其相关机制。方法:Western blot法检测卵巢癌细胞株A2780、SKOV3及顺铂耐药细胞株A2780/DDP、SKOV3/DDP中CTHRC1蛋白表达水平。慢病毒lenti-sh CTHRC1转染SKOV3/DDP细胞,下调CTHRC1表达水平。CCK-8法检测沉默CTHRC1后SKOV3/DDP细胞增殖情况及顺铂半数抑制浓度(IC_(50))的变化。Western blot法检测转染后SKOV3/DDP细胞中抗凋亡蛋白Bcl-2的表达情况及Akt、STAT3的磷酸化水平。结果:与A2780和SKOV3细胞相比,顺铂耐药细胞株A2780/DDP和SKOV3/DDP细胞中CTHRC1蛋白表达水平相对较高(P<0.05)。靶向下调CTHRC1表达后,SKOV3/DDP-sh CTHRC1细胞的增殖能力无明显变化,但顺铂的IC_(50)显著降低(P<0.01),Akt、STAT3磷酸化水平受到抑制,Bcl-2表达水平降低(P<0.05)。结论:CTHRC1可能通过Akt、STAT3信号通路,调节抗凋亡蛋白Bcl-2的表达,继而影响卵巢癌细胞对顺铂的敏感度。
Objective: To investigate the effect of CTHRC1 on cisplatin-resistant ovarian cancer cells and its related mechanisms. Methods: Western blot was used to detect the expression of CTHRC1 protein in ovarian cancer cell lines A2780, SKOV3 and cisplatin-resistant cell lines A2780 / DDP and SKOV3 / DDP. Lentiviral lenti-sh CTHRC1 transfected SKOV3 / DDP cells, down regulation of CTHRC1 expression levels. The proliferation of SKOV3 / DDP cells and the half inhibitory concentration (IC 50) of CTHRC1 were detected by CCK-8 assay. Western blot was used to detect the expression of anti-apoptotic protein Bcl-2 and the phosphorylation of Akt, STAT3 in SKOV3 / DDP cells. Results: Compared with A2780 and SKOV3 cells, CTHRC1 protein expression was significantly higher in cisplatin-resistant cell lines A2780 / DDP and SKOV3 / DDP cells (P <0.05). After CTHRC1 was down-regulated, the proliferation of SKOV3 / DDP-sh CTHRC1 cells did not change significantly, but the IC50 of cisplatin was significantly decreased (P <0.01) and the phosphorylation of Akt and STAT3 was inhibited. The expression of Bcl-2 The level was lower (P <0.05). CONCLUSION: CTHRC1 may regulate the expression of anti-apoptotic protein Bcl-2 through Akt and STAT3 signaling pathways and subsequently affect the sensitivity of ovarian cancer cells to cisplatin.