目的 探討虎杖苷調(diào)節(jié)蛋白激酶B/哺乳動物雷帕霉素靶蛋白（Akt/mTOR）通路對胰腺癌細(xì)胞凋亡和自噬的影響。方法 將人胰腺癌PANC-1細(xì)胞分為對照組、虎杖苷（100、200、300μmol/L）組、SC79組、虎杖苷+SC79組。CCK-8法檢測細(xì)胞活力；流式細(xì)胞術(shù)檢測細(xì)胞凋亡；單丹磺酰尸胺（MDC）染色檢測細(xì)胞中自噬小體陽性率；免疫熒光染色檢測細(xì)胞中微管相關(guān)蛋白1輕鏈3(LC3)陽性率；q RT-PCR檢測細(xì)胞中Bcl-2關(guān)聯(lián)X蛋白（Bax）、天冬氨酸特異性半胱氨酸蛋白酶-3(Caspase-3)、Beclin1、p62 mRNA表達(dá)；Western blotting檢測細(xì)胞中LC3、Beclin1、p62、p-Akt、p-mTOR蛋白相對表達(dá)量。結(jié)果 與對照組比較，虎杖苷各劑量組PANC-1細(xì)胞活力、p62蛋白和m RNA相對表達(dá)水平、p-Akt/Akt、p-mTOR/mTOR顯著降低，細(xì)胞凋亡率、自噬小體陽性率、LC3陽性率、Bax、Caspase-3、Beclin1 mRNA表達(dá)及LC3-II/LC3-I、Beclin1蛋白相對表達(dá)量顯著升高，且呈劑量相關(guān)性（P<0.05）。與虎杖苷300μmol/L組比較，虎杖苷+SC79組PANC-1細(xì)胞活力、p62蛋白和m RNA相對表達(dá)水平、p-Akt/Akt、p-mTOR/mTOR顯著升高，細(xì)胞凋亡率、自噬小體陽性率、LC3陽性率、Bax、Caspase-3、Beclin1 m RNA表達(dá)及LC3-II/LC3-I、Beclin1蛋白相對表達(dá)水平顯著降低（P<0.05）。結(jié)論 虎杖苷誘導(dǎo)PANC-1細(xì)胞自噬與凋亡的機(jī)制可能與抑制Akt/mTOR通路有關(guān)。

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

河南省科技攻關(guān)項(xiàng)目(222102310725)