白花蛇舌草-半枝莲药对组分对胃癌SGC-7901细胞增殖、线粒体自噬及凋亡的影响

Effects of Herba Hedyotis and Herba Scutellariae Barbatae Couplet Medicines Component on the Proliferation,Autophagy and Apoptosis in Gastric Cancer SGC-7901 Cells

目的:观察白花蛇舌草-半枝莲药对组分对胃癌SGC-7901细胞增殖、线粒体自噬及凋亡的影响。方法:实验分为对照组(正常培养SGC-7901细胞),白花蛇舌草-半枝莲药对组分低剂量组(SGC-7901细胞+25 mg·L^-1)、中剂量组(SGC-7901细胞+37.5 mg·L^-1)、高剂量组(SGC-7901细胞+50 mg·L^-1)。噻唑蓝比色法检测各组细胞培养72 h后细胞增殖抑制率;透射电镜观察各组细胞超微结构;Lyso-Tracker Red荧光染色观察SGC-7901细胞自噬情况;荧光TUNEL染色观察SGC-7901细胞凋亡情况;JC-1染色检测SGC-7901细胞线粒体膜电位变化;Western blot检测各组细胞BNIP3、Beclin-1、LC3Ⅰ/LC3Ⅱ、Cyt C、Caspase-8、Caspase-9、Caspase-3蛋白表达。结果:与对照组比较,白花蛇舌草-半枝莲药对组分各剂量组均显著抑制胃癌SGC-7901细胞增殖(P<0.01)。与对照组比较,白花蛇舌草-半枝莲药对组分各剂量组细胞均出现细胞线粒体膜电位降低、线粒体自噬及细胞凋亡现象(P<0.01),BNIP3、Beclin-1、Cyt C、Caspase-8、Caspase-9、Caspase-3蛋白表达增加(P<0.01),LC3Ⅰ/LC3Ⅱ比值降低(P<0.01)。结论:白花蛇舌草-半枝莲药对组分对胃癌SGC-7901细胞增殖有显著抑制作用,其机制可能与激活线粒体自噬的同时,激活线粒体凋亡信号通路,诱导肿瘤细胞凋亡有关。

Objective:To observe the effect of Herba Hedyotis and Herba Scutellariae Barbatae couplet medicines component on the proliferation,autophagy and apoptosis in gastric cancer SGC-7901 Cells.Methods:The experiment was divided into control group(SGC-7901 cells were cultured normally),Herba Hedyotis and Herba Scutellariae Barbatae couplet medicines group which was di-vided into low dose group(SGC-7901 cells+25 mg·L^-1),medium dose group(SGC-7901 cells+37.5 mg·L^-1),high dose group(SGC-7901 cells+50 mg·L^-1).Thiazole blue colorimetry was used to detect the inhibition rate of cell proliferation in each group after 72 hours of culture;transmission electron microscopy was used to observe the ultrastructure of each group;Lyso-Tracker Red fluorescence staining was used to observe the autophagy of SGC-7901 cells;fluorescence TUNEL staining was used to observe the apoptosis of SGC-7901 cells;JC-1 staining was used to detect the changes of mitochondrial membrane potential of SGC-7901 cells;the protein expression of BNIP3,beclin-1,LC3Ⅰ/LC3Ⅱ,Cyt C,Caspase-8,caspase-9 and caspase-3 were detected by west-ern blot.Results:Compared with the control group,each dose groups of Herba Hedyotis and Herba Scutellariae Barbatae couplet medicines component significantly inhibited the proliferation of gastric cancer SGC-7901 Cells(P<0.01).Compared with the control group,the cell membrane potential,mitochondrial autophagy and apoptosis were decreased in each dose group(P<0.01),the protain expression of BNIP3,beclin-1,Cyt C,Caspase-8,Caspase-9,Caspase-3 was increased(P<0.01),and the ratio of LC3Ⅰ/LC3Ⅱwas decreased(P<0.01).Conclusion:Herba Hedyotis and Herba Scutellariae Barbatae couplet medicines component can inhibit the proliferation of gastric cancer SGC-7901 cells.The mechanism may be related to activating mitochondrial autoph-agy,activating mitochondrial apoptosis signal pathway and inducing tumor cell apoptosis.