安罗替尼在宫颈癌HeLa/DDP细胞中的作用及逆转顺铂耐药性机制研究

Study on the role of Anlotinib in cervical cancer HeLa/DDP cells and the mechanism of reversing Cisplatin resistance

目的:探究安罗替尼对HeLa/DDP(宫颈癌顺铂耐药细胞)细胞增殖和凋亡的影响及其逆转顺铂耐药的机制。方法:不同浓度顺铂及安罗替尼处理HeLa/DDP细胞,CCK-8法检测细胞增殖能力;流式细胞术检测细胞凋亡;Western Blot检测P-gp的表达水平差异。结果:顺铂对HeLa/DDP细胞的增殖具有抑制作用,并呈剂量及时间依赖性。安罗替尼对HeLa/DDP细胞的增殖抑制作用呈浓度依赖性。安罗替尼、安罗替尼联合顺铂可以抑制HeLa/DDP细胞增殖(P<0.05)。流式细胞术结果发现安罗替尼组及安罗替尼联合顺铂组细胞凋亡率均升高(P<0.05)。安罗替尼联合顺铂显著抑制细胞增殖,诱导凋亡。Western Blot提示安罗替尼联合顺铂组与对照组和顺铂组及安罗替尼单药组相比,P-gp蛋白表达上调(P>0.05)。结论:安罗替尼可抑制宫颈癌HeLa/DDP细胞增殖,诱导凋亡;安罗替尼联合顺铂抑制细胞增殖、诱导凋亡作用更显著;安罗替尼可增加宫颈癌HeLa/DDP顺铂耐药株对顺铂的敏感性,逆转耐药性,但不能介导耐药相关蛋白P-gp表达下调。

Objective:To investigate the effects of Anlotinib on the proliferation and apoptosis of cervical cancer HeLa/DDP cells and its mechanism of reversing Cisplatin resistance.Methods:HeLa/DDP cells were treated with different concentrations of Cisplatin and Anlotinib,and the proliferation ability of cells was detected by CCK-8 method.Apoptosis was detected by flow cytometry and the difference of P-gp expression level was detected by Western Blot.Results:Cisplatin inhibited the proliferation of HeLa/DDP cells in a dose-and time-dependent manner.Anlotinib inhibited the proliferation of HeLa/DDP cells in a concentration-dependent manner.Anlotinib and Anlotinib combined with Cisplatin inhibited the proliferation of HeLa/DDP cells(P<0.05).Flow cytometry results revealed an increased apoptosis rate in both the Anlotinib group and the Anlotinib combined with Cisplatin group(P<0.05).Anlotinib combined with Cisplatin significantly inhibited cell proliferation and induced apoptosis.Western Blot suggested that P-gp protein expression was upregulated in the Anlotinib combined with Cisplatin group compared with the control group and the Cisplatin group and the Anlotinib monotherapy group( P >0.05). Conclusion: Anlotinib inhibits proliferation and induces apoptosis in cervical cancer HeLa/DDP.Anlotinib combined with Cisplatin inhibits cell proliferation and induces apoptosis more significantly.Anlotinib increases the sensitivity of Cisplatin-resistant strains of cervical cancer HeLa/DDP to Cisplatin and reverses drug resistance,but does not mediate the down-regulation of resistance-associated protein P-gp expression.