二甲双胍抑制人MDA-MB-231细胞增殖及迁移的体外实验研究

目的： 研究二甲双胍对体外培养的人三阴性乳腺癌MDA-MB-231细胞增殖及迁移的影响。方法： MTT实验分为4个处理组，分别为RPMI 1640培养基对照组，浓度为1、2、4 mmol/L的二甲双胍处理组，在作用48、72 h后测定各组的细胞活力并进行统计学分析；倒置相差显微镜下观察药物处理72 h后人乳腺癌MDA-MB-231细胞与对照组相比的形态变化；Giemsa染色观察4 mmol/L二甲双胍作用48 h后的细胞形态；采用细胞划痕实验和Transwell实验，检测二甲双胍（2、4 mmol/L）作用人乳腺癌MDA-MB-231细胞24 h后，对细胞迁移能力的影响。结果： MTT实验结果显示4 mmol/L二甲双胍作用72 h可显著抑制MDA-MB-231细胞的活力，抑制率为（29.83±2.25）％，与对照组相比，细胞活力降低（P<0.05）。倒置相差显微镜下形态学观察发现，与对照组相比，4 mmol/L二甲双胍处理组细胞密度降低，变圆的细胞数量增加，细胞与细胞之间的联系减少。Giemsa染色结果显示4 mmol/L二甲双胍作用48 h后，部分细胞出现凋亡细胞核碎裂形态。划痕实验结果表明，2、4 mmol/L二甲双胍作用24 h后细胞汇合度明显低于对照组，其中4 mmol/L处理组的划痕愈合率为（52.67±4.48）%，与对照组间的差异显著（P<0.01）；Transwell实验结果表明，2、4 mmol/L二甲双胍作用细胞24 h后穿膜细胞数量减少，穿膜细胞数量分别为（61.6±1.6）、（51.3±2.6）个，均低于对照组的（99.3±18.9）个（P<0.05）。结论： 二甲双胍在体外可抑制人乳腺癌MDA-MB-231细胞的增殖及迁移。

Metformin inhibitd proliferation and migration of human breast cancer cell in vitro

OBJECTIVE: To investigate effects of metformin on proliferation and migration of human breast cancer cell MDA-MB-231 in vitro. METHODS: Cell cultures were divided into four treatment groups:RPMI 1640 medium group and metformin treatment groups (1,2,4 mmol/L). Cell viability was measured at 48 and 72 h after treatment. Morphological changes of cells were observed under inverted microscopes after cells were stained with Giemsa. Effects of metformin on cell migration were detected by wound healing and Transwell assays. RESULTS: The MTT assay showed that metformin at 4 mmol/L significantly inhibited cell viability l for 72 h,and the inhibition rate was (29.83±2.25%). Compared with the control group,the decrease of cell viability was statistically significant (P<0.05). In the 4 mmol/L treatment group,the number of round cells increased,the cell density decreased obviously,and the connection between cells decreased. After treatment with metformin at 4 mmol/L for 48 hours,the cells showed obvious nuclear fragmentation. Wound healing experiment showed that the cell confluence of 2,4 mmol/L treatment groups were obviously lower than that of the control group after 24 h. The wound healing rate of 4 mmol/L treatment group was (52.67±4.48%),which was significantly different from that of the control group (P<0.01). Transwell experiments showed that the number of perforating cells decreased after treated with metformin at the concentration of 2,4 mmol/L for 24 h. The numbers of perforating cells were (61.6±1.6) and (51.3±2.6),which was significantly different from that of the control group (99.3±18.9) (all P<0.05). CONCLUSION: Metformin inhibited proliferation and migration of human breast cancer cell in vitro.