蟾蜍灵诱导人宫颈癌C33A细胞自噬作用的机理研究

目的:探讨蟾蜍灵诱导人宫颈癌C33A细胞自噬的作用。方法:不同浓度梯度的蟾蜍灵处理人宫颈癌C33A细胞，选用CCK－8法检测蟾蜍灵对人宫颈癌C33A细胞的杀伤作用。透射电镜观察给药后C33A细胞的自噬现象。流式细胞仪检测细胞内活性氧（ROS）水平，蛋白免疫印迹法检测自噬LC3蛋白表达及相关信号通路JNK、p-JNK蛋白表达。结果:蟾蜍灵对人宫颈癌C33A细胞的生长抑制作用呈时间-剂量依赖性。蟾蜍灵给药后可诱导C33A细胞发生自噬，蛋白印迹实验结果表明LC3蛋白表达随药物浓度升高而增加。蟾蜍灵诱导C33A细胞自噬发生与其促ROS水平升高激活了JNK信号通路有关。结论:蟾蜍灵诱导C33A细胞自噬的机制可能与通过ROS/JNK通路促进细胞自噬有关，以此发挥其抗肿瘤的作用。

Study on the autophagy mechanism of C33A cells induced by bufalin in human cervical cancer

Objective:To investigate the autophagy effect of bufalin on C33A cells induced by cervical cancer.Methods:Different concentrations of bufalin were used to treat human cervical cancer C33A cells,and CCK-8 method was used to detect the killing effect of bufalin on human cervical cancer C33A cells.The autophagy of C33A cells was observed by transmission electron microscopy.The reactive active oxygen (ROS) level was detected by flow cytometry,and the expression of autophagy LC3 protein and related signaling pathways JNK and p-JNK protein were detected by protein Western-blot.Results:The growth inhibition effect of bufalin on human cervical cancer C33A cells was time-dose dependent.Bufalin can induce autophagy in C33A cells after bufalin treatment,and the results of Western-blot experiment showed that the expression of LC3 protein increased with the increase of drug concentration.The autophagy of C33A cells induced by bufalin is related to the activation of JNK signaling pathway by the increased ROS level.Conclusion:The mechanism of bufalin inducing autophagy in C33A cells may be related to promoting autophagy through ROS/JNK pathway,so as to exert its anti-tumor effect.