目的 探讨虎杖苷调节蛋白激酶B/哺乳动物雷帕霉素靶蛋白（Akt/mTOR）通路对胰腺癌细胞凋亡和自噬的影响。方法 将人胰腺癌PANC-1细胞分为对照组、虎杖苷（100、200、300μmol/L）组、SC79组、虎杖苷+SC79组。CCK-8法检测细胞活力；流式细胞术检测细胞凋亡；单丹磺酰尸胺（MDC）染色检测细胞中自噬小体阳性率；免疫荧光染色检测细胞中微管相关蛋白1轻链3(LC3)阳性率；q RT-PCR检测细胞中Bcl-2关联X蛋白（Bax）、天冬氨酸特异性半胱氨酸蛋白酶-3(Caspase-3)、Beclin1、p62 mRNA表达；Western blotting检测细胞中LC3、Beclin1、p62、p-Akt、p-mTOR蛋白相对表达量。结果 与对照组比较，虎杖苷各剂量组PANC-1细胞活力、p62蛋白和m RNA相对表达水平、p-Akt/Akt、p-mTOR/mTOR显著降低，细胞凋亡率、自噬小体阳性率、LC3阳性率、Bax、Caspase-3、Beclin1 mRNA表达及LC3-II/LC3-I、Beclin1蛋白相对表达量显著升高，且呈剂量相关性（P<0.05）。与虎杖苷300μmol/L组比较，虎杖苷+SC79组PANC-1细胞活力、p62蛋白和m RNA相对表达水平、p-Akt/Akt、p-mTOR/mTOR显著升高，细胞凋亡率、自噬小体阳性率、LC3阳性率、Bax、Caspase-3、Beclin1 m RNA表达及LC3-II/LC3-I、Beclin1蛋白相对表达水平显著降低（P<0.05）。结论 虎杖苷诱导PANC-1细胞自噬与凋亡的机制可能与抑制Akt/mTOR通路有关。

Objective To investigate the effects of polydatin on apoptosis and autophagy of pancreatic cancer cells by regulating Akt/mTOR pathway. Methods Human pancreatic cancer PANC-1 cells were separated into control group, polydatin（100, 200, 300 μmol/L） group, SC79 group, polydatin + SC79 group. CCK-8 method was applied to detect cell viability. Flow cytometry was applied to detect cell apoptosis. Monodansylcadaverine（MDC） staining was applied to detect the positive rate of autophagosomes in cells. Immunofluorescence staining was applied to detect the positive rate of microtubule associated protein 1 light chain 3（LC3） in cells. qRT-PCR was applied to detect the mRNA expression of Bax, Caspase-3, Beclin1, and p62 in cells. Western blotting was applied to detect LC3, Beclin1, p62, p-Akt, and p-mTOR proteins in cells. Results Compared with the control group, the PANC-1 cell viability, p62 mRNA expression, and relative expression level of p62 protein, p-Akt/Akt, p-mTOR/mTOR were lower in polydatin groups, the apoptosis rate, autophagosome positivity rate, LC3 positive rate, Bax, Caspase-3, Beclin1 mRNA expression, and LC3-II/LC3-I, the relative expression level of Beclin1 protein were higherin a dose-dependent manner（P＜0.05）. Compared with polydatin 300 μmol/L group, the PANC-1 cell viability, p62 mRNA expression, and relative expression level of p62 protein, p-Akt/Akt, p-mTOR/mTOR in the polydatin + SC79 group were higher, the apoptosis rate, autophagosome positivity rate, LC3 positive rate, Bax, Caspase-3, and Beclin1 m RNA expression, and LC3-II/LC3-I, the relative expression level of Beclin1 protein were lower（P＜0.05）. Conclusion The mechanism by which polydatin induces autophagy and apoptosis in PANC-1 cells may be related to the inhibition of the Akt/mTOR pathway.

R285.5

河南省科技攻关项目(222102310725)