养正散结汤对人胃癌癌前病变细胞增殖、凋亡及抑癌性miRNA表达的影响

Yangzheng Sanjie Decoction Regulates Proliferation and Apoptosis of Gastric Precancerous Cells by Enhancing Onco-suppressor miRNA Expression

目的探讨养正散结汤对人胃癌癌前病变(PLGC)细胞增殖、凋亡及抑癌性miRNA表达的影响。方法采用N-甲基-N′-硝基-N-亚硝基胍(MNNG)转化的人胃黏膜上皮细胞GES-1(MC细胞)作为PLGC细胞模型,以10%浓度的养正散结汤含药血清处理MC细胞,采用CCK-8法检测细胞增殖,Annexin V-FITC/PI双染法流式细胞术检测细胞凋亡,实时定量PCR检测let-7a和miR-7的表达。结果养正散结汤组48、72 h的OD值均低于阴性对照组(P<0.01),细胞凋亡率高于阴性对照组(P<0.01);MC细胞let-7a、miR-7的表达均低于GES-1细胞(P<0.05);经养正散结汤大鼠含药血清干预后,let-7a、miR-7的表达显著上调(P<0.01)。结论养正散结汤可能通过上调抑癌性miRNA let-7a、miR-7的表达而抑制MC细胞增殖、诱导其凋亡。

Objective To explore effect of Yangzheng Sanjie Decoction （YZSJD） on proliferation, apoptosis and onco-suppressor miRNA mediating in gastric precancerous cells. Methods The N-methyl-N＇-nitro-N-nitrosoguanidine （MNNG） transformed human gastric epithelial cell line GES-1 （MC） was used as an in vitro precancerous lesions of gastric cancer （PLGC） model. MC cells were cultured with 10% of YZSJD-containing rat serum （YZSJD group） or non-medicated rat serum （control group） respectively. The proliferation of cells was measured using Cell Counting Kit-8 assay, cell apoptosis rate were detected by Annexin V-FITC/PI double staining flow cytometry analysis, and the expression of let-7a and miR-7 was analyzed by real time quantitative PCR. Results The OD value in YZSJD group was significantly lower than that in control group （P 〈 0.01 ） while the total apoptosis rate was higher （P 〈 0.01 ）. The expression levels of let-7a and miR-7 in MC cells were lower than that in GES-1 cells （P 〈 0.05 ）. After being treated with YZSJD-containing serum, the levels of let-7a and miR-7 were both up-regulated（P 〈 0.01 ）. Conclusion YZSJD inhibits proliferation and induces apoptosis by up-regulating let-7a and miR-7 in MC cells.