二甲双胍联合顺铂对人成骨肉瘤MG-63细胞增殖的抑制作用  

Inhibition of Metformin Combined with Cisplatin on Proliferation of Human Osteosarcoma MG-63 Cells

作　　者：张翔 赵帅帅 王雅玲 张洁[3] 殷松娜 吴博[5] ZHANG Xiang;ZHAO Shuai-Shuai;WANG Ya-Ling;ZHANG Jie;YIN Song-Na;WU Bo(Department of Medical Genetics&Cell Biology,Medical School of Yan'an University,Yan'an 716000,Shaanxi,China;Key Laboratory of Neuroscience in Yan'an,Medical School of Yan'an University,Yan'an 716000,Shaanxi,China;Department of Anatomy,Medical School of Yan'an University,Yan'an 716000,Shaanxi,China;Department of Physiology,Medical School of Yan'an University,Yan'an 716000,Shaanxi,China;Department of Histology and Embryology,Medical School of Yan'an University,Yan'an 716000,Shaanxi,China)

机构地区：[1]延安大学医学院遗传学与细胞生物学教研室,陕西延安716000 [2]延安大学医学院延安市神经科学重点实验室,陕西延安716000 [3]延安大学医学院解剖学教研室,陕西延安716000 [4]延安大学医学院生理学教研室,陕西延安716000 [5]延安大学医学院组织胚胎学教研室,陕西延安716000

出　　处：《中国生物化学与分子生物学报》2021年第7期960-966,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：陕西省教育厅项目(No.19JK0958);陕西省科技厅项目(No.2020JQ-800,No.2021JQ-639);延安大学校级项目(No.YDQ2019-38)资助。

摘　　要：为研究二甲双胍联合顺铂处理对人成骨肉瘤MG-63细胞的影响及潜在作用机制,本研究使用无药物处理组为对照组细胞,使用二甲双胍和顺铂联合处理MG-63细胞,采用CCK8法和流式细胞术检测细胞的增殖凋亡情况;用细胞克隆形成实验检测各组克隆形成率;用Trans well实验检测各组细胞迁移侵袭能力,用qPCR法和蛋白质印迹法检测凋亡相关基因的RNA水平和蛋白质水平表达量。结果显示,二甲双胍联合顺铂处理对人成骨肉瘤MG-63细胞的增殖抑制作用更加显著(P<0.01);流式细胞术检测结果也表明,二甲双胍联合顺铂处理能够更显著的促进人成骨肉瘤MG-63细胞的凋亡(P<0.01);细胞克隆形成的结果表明,二甲双胍联合顺铂处理抑制人成骨肉瘤MG-63细胞克隆的形成(P<0.01);Trans well结果表明,二甲双胍联合顺铂处理抑制人成骨肉瘤MG-63细胞的迁移和侵袭能力(P<0.01);qPCR和蛋白质印迹法检测结果表明,二甲双胍联合顺铂处理后可下调人成骨肉瘤MG-63细胞中凋亡相关基因MCl-1和XIAP(P<0.01),上调凋亡标志基因CASPASE-3和Cyto C(P<0.01),及细胞迁徙侵袭相关基因MMP-2和MMP-9(P<0.01)表达水平。本研究结果表明,二甲双胍联合顺铂对人成骨肉瘤MG-63细胞增殖的抑制可能是通过调节MCL-1和XIAP来促进人成骨肉瘤MG-63细胞的凋亡,通过调节MMP-2和MMP-9途径来实现抑制人成骨肉瘤MG-63细胞的迁移侵袭。In order to study the effect and its potential mechanism of metformin combined with cisplatin treatment on human osteosarcoma MG-63 cells,MG-63 cells were treated with metformin and cisplatin and the cell proliferation and apoptosis were detected using CCK8 and flow cytometry;Cell clone formation experiment was performed to detect clone formation rate in each group;Trans well experiment was used to detect the migration and invasion ability of osteosarcoma MG-63 cells,and qPCR and Western blot were used to detect the expression of RNA and protein of apoptosis-related genes.The results showed that metformin combined with cisplatin inhibited the proliferation of human osteosarcoma MG-63 cells and promoted their apoptosis significantly(P<0.01);Metformin combined with cisplatin inhibited the clone formation(P<0.01),and the migration and invasion of human osteosarcoma MG-63 cells(P<0.01);Furthermore,metformin combined with cisplatin down-regulated the expression of apoptosis-related genes MCl-1 and XIAP(P<0.01),but up-regulated the expression of apoptosis-related genes CASPASE-3 and Cyto C(P<0.01)and migration and invasion related genes MMP-2 and MMP-9(P<0.01).Our study indicated that metformin combined with cisplatin inhibited proliferation,promoted cell apoptosis through MCl-1 and XIAP,and inhibited cell migration and invasion by regulating the MMP-2 and MMP-9 pathways in human osteosarcoma MG-63 cells.

关 键 词：骨肉瘤 顺铂 二甲双胍 增殖 

分 类 号：R73-3[医药卫生—肿瘤]