摘要
目的研究川芎嗪(TMP)对人胃癌SGC-7901细胞增殖的抑制作用并探讨其机制。方法分别以DMSO(空白对照组),TMP低、中、高剂量(200、400、800μg/ml)和LY294002(PI3K特异性抑制剂,5μg/ml)干预对数生长期SGC-7901细胞48 h。采用MTT法检测SGC-7901细胞增殖抑制率,Annexin V-FITC/PI流式细胞术检测细胞凋亡水平,吖啶橙染色法观察细胞自噬状况,Western blot法检测p-PI3K、p-Akt、p-m TOR、Cleaved Caspase-9、Cleaved Caspase-3、LC3蛋白表达,计算LC3-Ⅱ/LC3-I比值。结果与空白对照组比较,经TMP低、中、高剂量或LY294002干预能够明显提高SGC-7901细胞增殖抑制率,诱导细胞凋亡并显著提高凋亡指数,明显增多自噬溶酶体数量;经TMP中、高剂量或LY294002干预能够明显下调p-PI3K、p-Akt、p-m TOR蛋白表达,上调Cleaved Caspase-9、Cleaved Caspase-3蛋白表达,提高LC3-Ⅱ/LC3-I比值(P<0.05,P<0.01),差异有统计学意义。与LY294002组比较,经TMP高剂量干预能够明显提高SGC-7901细胞增殖抑制率和凋亡指数、增多自噬溶酶体数量;下调p-PI3K、p-Akt、p-m TOR表达并上调Cleaved Caspase-3表达,提高LC3-Ⅱ/LC3-I比值(P<0.05,P<0.01),差异有统计学意义。结论 TMP能够通过诱导人胃癌SGC-7901细胞凋亡和自噬抑制其增殖,可能与抑制PI3K/Akt/m TOR通路活化进而影响其下游凋亡和自噬相关蛋白表达有关。
Objective To investigate the effect of Tetramethylpyrazine(TMP) on inhibiting the proliferation of human gastric cancer SGC-7901 cells and its molecular mechanism.Methods The DMSO(blank control group),TMP low,medium,high dose(200,400,800 μg/ml) and LY294002(PI3 K specific inhibitor,5μg/ml) were used to interfere with SGC-7901 cells in logarithmic growth respectively for 48 h.The proliferation inhibition rate of SGC-7901 cells was detected by MTT method.The analysis of apoptotic status were detected by Annexin V-FITC/PI staining flow cytometry.The autophagy of cells was observed by acridine orange staining.The expression of p-PI3K,p-Akt,p-m TOR,Cleaved Caspase-3,Cleaved Caspase-9,LC3 proteins were detected by Western blot,the ratio of LC3-Ⅱ/LC3-I were calculated.Results Compared with blank control group,intervented by TMP low,medium,high dose or LY294002 could increase the proliferation inhibition rate,apoptosis rate and the number of autophagic vesicles;intervented by TMP medium,high dose or LY294002 could down-regulate the expression of p-PI3 K,p-Akt,p-m TOR and up-regulate the expression of Cleaved Caspase-3/-9,increase the ratio of LC3-Ⅱ/LC3-I;all of the difference were significant(P<0.05 or P<0.01).Compared with LY294002 group,intervented by TMP high dose could increase the proliferation inhibition rate,increase the apoptosis rate and the number of autophagic vesicles;down-regulate the expression of p-PI3 K,p-Akt,p-m TOR and up-regulate the expression of Cleaved Caspase-3,increase the ratio of LC3-Ⅱ/LC3-I;all of the difference were significant(P<0.05 or P<0.01).Conclusions TMP can inhibit the proliferation of human gastric cancer SGC-7901 cells by inducing apoptosis and autophagy;which mechanism may be related to inhibiting PI3 K/Akt/m TOR signaling pathway and further affecting downstream apoptosis and autophagy related protein expression.
作者
霍浩然
秦瑞峰
薛佳栋
袁增江
HUO Hao-ran;QIN Rui-feng;XUE Jia-dong;YUAN Zeng-jiang(Third Department of General surgery,Handan Central Hospital,Handan Hebei 056001)
出处
《世界中西医结合杂志》
2020年第10期1819-1823,共5页
World Journal of Integrated Traditional and Western Medicine
基金
河北省医学科学研究课题计划(20191845)。
