鸡屎藤苷酸诱导SGC-7901胃癌细胞凋亡并抑制细胞恶性增殖和侵袭

Paederosidic Acid Induces Apoptosis of Gastric Cancer SGC-7901 Cells and Inhibits Their Malignant Proliferation and Invasion

目的:初步探索鸡屎藤苷酸(PA)诱导SGC-7901胃癌细胞凋亡并抑制细胞恶性增殖和侵袭的可能机制。方法:选取SGC-7901胃癌细胞为研究对象,利用CCK-8法进行浓度梯度实验确定后续实验浓度,利用流式细胞术检测0、20、40、80μg/mL的PA处理后SGC-7901胃癌细胞凋亡率以及线粒体膜电位变化情况,BrdU法和Transwell小室实验SGC-7901胃癌细胞的增殖活性以及侵袭能力,qRT-PCR检测抑癌基因P53、P21的mRNA相对表达水平,Western Blotting检测凋亡相关蛋白Bcl-2、Bax、Caspase-3、c-Myc和转移相关蛋白VEGF、波形蛋白表达水平。结果:CCK-8梯度浓度试验发现PA浓度从40μg/mL开始对SGC-7901胃癌细胞活性产生了明显抑制作用(P<0.05),后续以20、40、80μg/mL为试验浓度;与0μg/mL的PA处理比较,40、80μg/mL的PA处理后SGC-7901胃癌细胞凋亡率、Bax/Bcl-2、Cleaved-Caspase-3/Caspase-3蛋白表达水平以及P53、P21相对表达量升高,线粒体膜电位、细胞增殖活性、侵袭率、c-Myc、VEGF、波形蛋白表达水平降低,差异均有统计学意义(均P<0.05)。结论:PA可能通过降低线粒体膜电位促进线粒体凋亡途径诱导细胞凋亡,促进抗癌基因表达抑制细胞的恶性增殖,并抑制上皮间质转化降低侵袭能力。

Objective:To preliminarily explore the underlying mechanism of paederosidic acid(PA)in inducing the apoptosis of gastric cancer SGC-7901 cells and inhibiting their malignant proliferation and invasion.Methods:The concentration gradient test was conducted on SGC-7901 cells by CCK-8 to determine the subsequent experimental concentration.The changes in the apoptosis rate of SGC-7901 cells and mitochondrial membrane potential(MMP)after PA treatment(0,20,40,and 80μg/mL)were detected by flow cytometry.The proliferation activity and invasion ability of SGC-7901 cells were detected by BrdU staining and Transwell assay.The relative expression levels of tumor suppressor genes(P53 and P21)were detected by qRT-PCR.The expression levels of apoptosis-related proteins(Bcl-2,Bax,Caspase-3,and c-Myc)and metastasis-related proteins(VEGF and vimentin)were detected by Western blot.Results:As revealed by the CCK-8 concentration gradient test,PA at the concentration higher than 40μg/mL showed inhibitory effects on the activity of SGC-7901 cells(P<0.05).The subsequent experimental concentrations were determined as 20,40,and 80μg/mL.Compared with the results after PA treatment at 0μg/mL,the apoptosis rate of SGC-7901 cells,protein expression levels of Bax/Bcl-2 and cleaved-Caspase-3/Caspase-3,and relative mRNA expression levels of P53 and P21 increased,while MMP,cell proliferation activity,invasion rate,and protein expression levels of c-Myc,VEGF,and vimentin decreased after PA treatment at 40 and 80μg/mL(P<0.05).Conclusion:PA may induce cell apoptosis by reducing MMP and promoting mitochondrial apoptosis pathway to promote the expression of anti-oncogenes,inhibit malignant proliferation and epithelial-mesenchymal transition,and reduce the invasion ability.