二甲双胍联合奥沙利铂对结肠癌SW480细胞凋亡及细胞周期的影响  

Effects of metformin combined with oxaliplatin on apoptosis and cell cycle of colon cancer SW480 cells

作　　者：陈玉忠[1] 魏丽 杜军[1] 韩冰清 石磊 李雷[1] 马家驰 CHEN Yuzhong;WEI Li;DU Jun;HAN Bingqing;SHI Lei;LI Lei;MA Jiachi(Department of Surgical Oncology,First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China;Department of Gynecological Oncology,First Affiliated Hospital of Bengbu Medical College;Department of Surgery,Bengbu Medical College;Department of Surgery,Suzhou First People’s Hospital)

机构地区：[1]蚌埠医学院第一附属医院肿瘤外科,蚌埠233000 [2]蚌埠医学院第一附属医院肿瘤妇科 [3]蚌埠医学院外科教研室 [4]宿州市第一人民医院外科

出　　处：《山西医科大学学报》2020年第10期1028-1034,共7页Journal of Shanxi Medical University

基　　金：蚌埠医学院科技发展基金项目(BYKF1879)。

摘　　要：目的探讨二甲双胍(metformin,Met)联合奥沙利铂(oxaliplatin,Oxa)对结肠癌SW480细胞凋亡及细胞周期的影响。方法用MTT法检测分别应用二甲双胍(0,5,10,20,40 mmol/L)、奥沙利铂(0,5,10,20,40μmol/L)对SW480细胞的增殖抑制作用。PI单染法检测分别应用二甲双胍(0,5,10,20,40 mmol/L)、奥沙利铂(0,5,10,20,40μmol/L)及二甲双胍(10 mmol/L)联合奥沙利铂(10μmol/L)对SW480细胞凋亡的影响。流式细胞术检测二甲双胍(10 mmol/L)、奥沙利铂(10μmol/L)及两者联合作用时SW480细胞周期的变化。Western blot法检测二甲双胍(10 mmol/L)、奥沙利铂(10μmol/L)及两者联合作用SW480细胞凋亡及细胞周期相关蛋白的表达。结果单独应用二甲双胍、奥沙利铂作用于SW480细胞后细胞的增殖随浓度增加而降低,二甲双胍联合奥沙利铂与单药作用相比细胞增殖抑制作用明显(P<0.05)。二甲双胍、奥沙利铂分别处理SW480细胞,细胞凋亡率随着给药浓度的增高而增加,联合用药组与单药组相比,细胞凋亡率明显增加(P<0.05)。细胞周期检测显示与空白组及单药组相比,联合用药组SW480细胞在G0/G1期比例增加(P<0.05)。Western blot结果显示与空白组和单药组相比,联合用药组作用于SW480细胞后Cyclin D1及CDK4的表达降低,P21的表达增加(P<0.05)。促凋亡蛋白cleaved-caspase-3表达增加,抗凋亡蛋白Bcl-2的表达降低,差异具有统计学意义(P<0.05)。结论二甲双胍联合奥沙利铂可抑制结肠癌SW480细胞的增殖并诱导其凋亡,其机制可能与细胞周期的阻滞有关。Objective To investigate the effects of metformin(Met)combined with oxaliplatin(Oxa)on apoptosis and cell cycle of colon cancer SW480 cells.Methods MTT method was used to detect the inhibitory effect of Met(0,5,10,20,40 mmol/L),Oxa(0,5,10,20,40μmol/L)on the proliferation of SW480 cells.The apoptotic cell was quantified after treated with Met(0,5,10,20,40 mmol/L),Oxa(0,5,10,20,40μmol/L)or the combination of Met(10 mmol/L)and Oxa(10μmol/L)for 24 h by flow cytometry,respectively.The cell cycle of SW480 cells was analyzed on flow cytometer after treated with Met(10 mmol/L),Oxa(10μmol/L)or their combination.The expression of apoptosis-and cell cycle-related proteins in SW480 cells after exposed to Met(10 mmol/L),Oxa(10μmol/L)or their combination was detected by Western blot.Results The proliferation of SW480 cells was decreased in a concentration-dependent manner after treated with Met(0,5,10,20,40 mmol/L)or Oxa(0,5,10,20,40μmol/L),respectively.The apoptosis rate of SW480 cells was increased in a concentration-dependent manner after treated with Met and Oxa,and the apoptosis rate of SW480 cells treated with combination of metformin and oxaliplatin was significantly increased compared with that of Met or Oxa alone(P<0.05).The cell apoptotic rate of SW480 cells was increased in a concentration-dependent manner after treated with Met(0,5,10,20,40 mmol/L)and Oxa(0,5,10,20,40μmol/L),and the apoptotic rate was higher after the combined treatment with Met(10 mmol/L)and Oxa(10μmol/L)than that of Met or Oxa alone.Cell cycle assay revealed that compared with blank group,Met group and Oxa group,the proportion of SW480 cells was increased at G0/G1 phase in combination group(P<0.05).Western blot showed that compared with blank group,Met group and Oxa group,the expression levels of Cyclin D1 and CDK4 were reduced in combination group,while the expression of P21 was increased,meanwhile,the proapoptotic protein cleaved-caspase-3 expression was increased(P<0.05),while the anti-apoptotic protein Bcl-2 expression was decreased(a

关 键 词：二甲双胍 奥沙利铂 结肠癌 SW480细胞 凋亡 细胞周期 

分 类 号：R735.35[医药卫生—肿瘤]