二甲双胍通过阻断Src/STAT3信号通路诱导胃癌MGC-803细胞凋亡的实验研究  

Metformin Inducing the Apoptosis of Gastric Cancer MGC-803 Cells by Blocking Scr/STAT3 Pathway

作　　者：谢静静[1] 张金花[1] 金剑英[1] 金丹[1] 郭群依[1] Xie Jingjing;Zhang Jinhua;Jin Jianying;Jin Dan;Guo Qunyi(Department of Oncology,Taizhou Hospital of Zhejiang Province,Zhejiang Taizhou 317000,China)

机构地区：[1]浙江省台州医院肿瘤内科,浙江台州317000

出　　处：《中国药师》2020年第2期241-245,251,共6页China Pharmacist

基　　金：2018年度台州恩泽医疗中心(集团)科研基金一般课题项目(编号:台恩泽医〔2018〕37号)。

摘　　要：目的:探讨二甲双胍通过阻断Src酪氨酸激酶(Src)/信号转导及转录激蛋白3(STAT3)信号通路诱导胃癌MGC-803细胞凋亡的机制。方法:设MGC803细胞组、氟尿嘧啶组(100.0μg·ml^-1)和二甲双胍低(80.0μg·ml^-1)、高(160.0μg·ml^-1)剂量组,各组细胞置于CO2培养箱(37℃、5%CO2、2%O2、93%N2)。以上各组每孔设6个平行样,培养72 h。培养结束后,细胞计数试剂盒-8(CCK-8)测定细胞活力,结晶紫染色测定单克隆形成数目,Transwell室测定侵袭能力,FACScan流式细胞仪测定细胞凋亡水平,实时荧光定量PCR(RT-PCR)及蛋白质印迹法(West Blot)测定p-SRC、p-STAT3 m RNA和蛋白水平。结果:氟尿嘧啶组和二甲双胍组的吸光度(A)值、存活率水平、克隆形成数目、穿膜数、p-SRC、p-STAT3 m RNA及蛋白表达水平显著低于MGC803细胞组(P<0.05),凋亡率显著高于MGC803细胞组(P<0.05)。二甲双胍低剂量组的A值、存活率水平、克隆形成数目、穿膜数、P-SRC、P-STAT3 m RNA、蛋白表达水平显著高于氟尿嘧啶组(P<0.05),凋亡率显著低于氟尿嘧啶组(P<0.05)。二甲双胍高剂量组与氟尿嘧啶组比较差异无统计学意义(P>0.05)。结论:二甲双胍能抑制胃癌MGC-803细胞增殖,促进胃癌MGC-803细胞凋亡;其机制与二甲双胍抑制P-SRC、P-STAT3 mRNA和蛋白的表达有关。Objective:To investigate the mechanism of metformin inducing the apoptosis of gastric cancer MGC-803 cells by blocking Src/STAT3 signaling pathway.Methods:MGC803 cell group,5-fluorouracil group(100.0μg·ml^-1),metformin low(80.0μg·ml^-1)and high(160.0μg·ml^-1)dose groups were set up.The cells in each group were placed in a CO2 incubator(37℃,5%CO2,2%O2,93%N2).The above groups were cultured for 72 hours with 6 parallel samples per hole.After the culture,cell viability kit-8(CCK-8)was used to measure the cell viability,crystal violet staining was used to determine the number of monoclonal formation,Transwell chamber was used to measure the invasive ability,FACScan flow cytometry was used to measure the apoptosis,and real-time quantitative PCR(RT-PCR)and Western blot were used to detect p-SRC,p-STAT3 m RNA and protein levels.Results:The absorbance(A)value,survival rate,number of clone formation,transmembrane number,p-SRC,p-STAT3 m RNA and protein expression levels in 5-fluorouracil group and metformin group were significantly lower than those in MGC803 cell group(P<0.05),and the apoptosis rate was significantly higher than that in MGC803 cell group(P<0.05).The A value,survival rate,number of clone formation,transmembrane number,P-SRC,P-STAT3 m RNA and protein expression levels in the low dose group of metformin were significantly higher than those in 5-fluorouracil group(P<0.05),and the apoptosis rate was significantly lower than that in MGC803 cell group(P<0.05).There were no significant differences between the high dose group of metformin and 5-fluorouracil group(P>0.05).Conclusion:Metformin can inhibit the proliferation and promote the apoptosis of gastric cancer MGC-803 cells.The mechanism is related to the inhibition of P-SRC and P-STAT3 m RNA and protein expression by metformin.

关 键 词：二甲双胍 Src酪氨酸激酶/信号转导及转录激蛋白3信号通路 胃癌 凋亡 

分 类 号：R965.1[医药卫生—药理学]