20(S)-原人参二醇对体外培养siha细胞自噬的影响

Effect of 20 (S)-proto-panaxdiol on autophagy of siha cells in vitro

目的:观察20(S)-原人参二醇(PPD)对人子宫颈癌siha细胞生长的抑制作用及其对自噬基因Beclin1及MAP1-LC3转录和表达的影响,探讨PPD抑制siha细胞增殖的机制。方法:体外培养人宫颈癌siha细胞分为阴性对照组(乙醇)、阳性对照组(40μmol·L-1顺铂)及PPD10、20和40μg·L-1组,分别用乙醇、顺铂和PPD处理细胞24、48、72和96h后,MTT法检测siha细胞增殖抑制率,RT-PCR、Western blotting及免疫细胞化学染色法观察各组siha细胞自噬相关基因Beclin1及MAP1-LC3的转录及蛋白表达情况。结果:与对照组比较,PPD10、20和40μg·L-1组细胞增殖抑制率明显增加(P<0.05),且随时间延长和剂量的增加而逐渐提高,自噬相关基因Beclin1及MAP1-LC3的转录水平及蛋白表达水平明显上调(P<0.01)。结论:PPD可抑制体外培养siha细胞增殖,诱导siha细胞发生自噬。其机制可能与上调自噬相关基因Beclin1及MAP1-LC3表达有关。

Objective To investigate the inhibitory effect of 20 （S）-proto-panaxdiol（PPD）on the growth of siha cells in vitro and detect the transcription and expressions of autophagy related gene Beclin 1and MAP1-LC3,and clarify the mechanism of its suppressing effect on siha cell proliferation.Methods The siha cells cultivated in vitro were treated with alcohol,40μmol·L-1 platinol（Pt）,10μg·L-1 PPD,20μg·L-1 PPD,40μg·L-1 PPD for 24,48,72and 96h,respectively.The inhibitory rate of cell proliferation in each group was detected by MTT, the transcription and expressions of autophagy related gene Beclin 1and MAP1-LC3were analyzed by RT-PCR, Western blotting and immunocytochemical staining.Results After treated with 10,20and 40μg·L-1 PPD,the inhibitory rates of siha cells were significantly increased compared with control group （P0.05）,and the inhibitory rate was increased gradually with the time prolongation of time and the increase of dose.The transcription and expressions of autophagy related gene Beclin 1and MAP1-LC3were significantly up-regulated（P0.01）. Conclusion PPD may suppress the proliferation of siha cells,and its mechanism may be related to up-regulating theexpressions of autophagy related gene Beclin 1and MAP1-LC3.