白藜芦醇对胃癌 SGC7901细胞自噬的影响及其机制探讨

摘要

Objective To study the influence of resveratrol ( RES) on the autophagy of SGC7901 cells and the role of classⅢPI3K.Methods SGC7901 cells were treated with different dose of RES (0, 10, 20, 50, 75, 100 μmol/L) for 24 h alone or pretreated with 3-MA ( or wortmannin, or chloroquine) for 1 h.Cell lysates were analyzed by Western blot for LC3-Ⅰ/LC3-Ⅱconversion;MDC staining was used to detect the morphologic changes of autophagy.Results The LC3-Ⅰ/LC3-Ⅱconversion and the number of autophagosomes increased in a dose dependent of RES and markedly increased in the presence of the protease inhibitors chloroquine.And the inhibitory function of class Ⅲ PI3K by its specific inhibitors ( wortmannin) had no impact on the level of autophagy induced by RES.Conclusion RES increased the autophagy of SGC7901 cells, and the autophagy was independent of classⅢPI3K.%目的：观察白藜芦醇（ RES）对胃癌SGC7901细胞自噬的影响，探讨Ⅲ型PI3 K激酶在其中的作用。方法不同浓度（0、10、20、50、75、100μmol/L） RES作用SGC7901细胞24 h，或Ⅲ型PI3K特异性抑制剂6-氨基-3-甲基嘌呤（3-MA）、渥曼青霉素（WM）及自噬晚期抑制剂氯喹（CQ）预处理细胞1 h后加入不同浓度RES再作用24 h；采用Western blot法观察自噬特异性蛋白LC3由Ⅰ型向Ⅱ型转化的情况，甲丹磺酰尸胺染色（ MDC）观察自噬囊泡。结果不同浓度RES均可引起LC3-Ⅰ向LC3-Ⅱ转化，且随浓度增加转化越明显；CQ预处理1 h后RES继续处理24 h，细胞内LC3蛋白转化、自噬酸性囊泡进一步增加；Ⅲ型PI3K特异性抑制剂3-MA（WM）不改变RES引起的LC3-Ⅰ向LC3-Ⅱ转化，不抑制MDC阳性细胞增加、细胞内酸性囊泡的聚集。结论 RES可增强胃癌SGC7901细胞内自噬水平，而非自噬降解功能削弱导致，该作用不依赖于Ⅲ型PI3K。