虎杖苷通过PI3KAKTmTOR信号通路诱导人宫颈癌细胞凋亡初步研究.docVIP

虎杖苷通过PI3KAKTmTOR信号通路诱导人宫颈癌细胞凋亡初步研究 　　[摘要] 通过观察虎杖苷对宫颈癌HeLa细胞体外生长的抑制作用，初步探讨其诱导凋亡的可能机制。不同浓度的虎杖苷（50，100，150 μmol?L-1）处理HeLa细胞后，采用MTT法检测虎杖苷对HeLa细胞增殖的抑制作用，AO/EB染色法荧光显微镜观察HeLa细胞凋亡的形态学变化；Annexin/PI双标记法检测HeLa细胞凋亡率；流式细胞仪分析HeLa细胞周期分布；RT-PCR和Western blot法检测HeLa细胞中PI3K，AKT，mTOR，P70S6K的mRNA和蛋白表达。结果显示，虎杖苷显著抑制HeLa细胞增殖，且具有一定剂量依赖性；虎杖苷能够引起HeLa细胞发生S期阻滞，促进细胞凋亡，显著下调HeLa细胞中PI3K，AKT，mTOR，P70S6K的mRNA和蛋白表达。表明虎杖苷具有抑制宫颈癌HeLa细胞增殖及诱导凋亡的作用，其机制可能与抑制PI3K/AKT/mTOR信号通路及其下游基因蛋白表达有关。 　　[关键词] 虎杖苷； 宫颈癌； HeLa细胞； PI3K/AKT/mTOR信号通路； 凋亡 　　[Abstract] To observe the effect of polydatin on proliferation and apoptosis of cervical cancer HeLa cells and explore its possible mechanism. The growth inhibitory effect was detected with MTT assay. After HeLa cells were treated with different concentrations （50， 100， 150 μmol?L-1） of polydatin， MTT assay was used to detect the inhibitory effect of polydatin on proliferation of HeLa cells； Acridine orange/ethidium bromide staining was used for morphological changes in apoptotic HeLa cells； Annexin/propidium iodide staining was applied to detect HeLa cell apoptotic rate. In addition， flow cytometry was employed to analyze apoptosis and cell cycle distribution； RT-PCR and Western blot assay were used to detect PI3K， AKT， mTOR， and P70S6K mRNA and protein expression levels. The results showed that polydatin significantly inhibited HeLa cells proliferation in a dose-dependent manner. Polydatin can cause S phase arrest for HeLa cells， promote cell apoptosis and decrease the mRNA and protein expression levels of PI3K， AKT， mTOR and P70S6K. It indicated that polydatin could inhibit proliferation and induce apoptosis of cervical cancer HeLa cells， and the mechanism may be associated with inhibiting the PI3K/AKT/mTOR signaling pathway and suppressing downstream gene expression. 　　[Key words] polydatin； cervical cancer； HeLa cell； PI3K/AKT/mTOR signaling pathway； apoptosis 　　?m颈癌是我国妇科最常见的恶性肿瘤，严重威胁广大妇女的生命健康。近年来发病率呈逐年上升趋势，且发病年龄不断年轻化。近年来对化疗在宫颈癌治疗中的应用进行了大量的研究，并取得了良好的效果[1]。常用的顺铂、五