二甲双胍对犬乳腺肿瘤细胞周期及其相关蛋白的影响  

Effect of metformin on cell cycle and the related proteins in canine mammary gland tumors

作　　者：王英雪[1] 刘佳[1] 杨超[1] 宋美英[1] 纪亚楠[1] 刘云[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《畜牧与兽医》2017年第4期111-115,共5页Animal Husbandry & Veterinary Medicine

基　　金：国家自然基金(31372492和31672617);国家重点研究项目(2016YFD0501008)

摘　　要：为研究二甲双胍对犬乳腺肿瘤细胞增殖和细胞周期的影响,以犬乳腺肿瘤CHMm和CHMp为模型,采用终浓度为0、5、10、20、40mmol/L二甲双胍对其进行干预,应用CCK-8法检测细胞增殖活性,流式细胞仪检测细胞周期分布比例,再通过Western blot检测细胞周期相关蛋白cyclin D1和p27的表达情况。二甲双胍作用48 h后,与对照组比较,CHMm细胞试验各组的成活率显著降低(P<0.05),而CHMp细胞除了5 mmol/L组(P>0.05),其他试验各组的成活率也显著低于对照组(P<0.05),并且均呈现一定的浓度依赖性。流式细胞仪分析细胞分布比例发现,CHMm的G0/G1期细胞比例从(38.7±5.89)%增加到(70.36±5.78)%,CHMp的G0/G1期细胞比例也从(37.03±4.23)%增加到(54.92±4.67)%。随着二甲双胍作用浓度的增加,CHMm和CHMp细胞内cyclin D1表达量呈现减少的趋势,而p27表达量呈现上升的趋势。研究结果表明,二甲双胍可以抑制犬乳腺肿瘤细胞体外增殖能力,诱导细胞发生G0/G1期阻滞,并下调cyclin D1和上调p27细胞周期相关蛋白表达量,从而为犬乳腺肿瘤治疗提供一定的理论基础。In order to determine the effect of metformin on cell proliferation and cell cycle in canine mammary gland tumors（CMGTs）,CHMm and CHMp were exposed to 0,2,5,10,20 and 40 m M final concentrations of metformin for 48 h.Cell viability was detected by CCK-8 assay,cell cycle was analysed by flow cytometry and cell cycle-related protein cyclin D1 and p27 were measured by western blot.After the treatment of metformin,compared with control（0 mmol/L）,cell viabilities of other concentration groups had a significant decrease（P0.05） in CHMm,CHMp viabilities also decreased significantly（P 0.05）,except for 5 mmol/L group（P 0.05）,and inhibition of metformin appeared concentration-dependent.Cell percentage at G0/G1 phase by flow cytometry rose from（37.03±4.23） % to（54.92±4.67） % in CHMm,from（37.03±4.23） % to（54.92 ± 4.67） % in CHMp,respectively.As for expressions of cyclin D1 and p27 in CHMm and CHMp,it is found that metformin could up-regulate the expression of cyclin D1 and down-regulate the expression of p27.In conclusion,metformin inhibited the proliferation of CMGTs cells,arrested cell cycle at G0/G1 phase and regulated the expression of cell cycle-related proteins,which lay a foundation for treatment of CMGTs.

关 键 词：二甲双胍 犬乳腺肿瘤 细胞周期 CYCLIN D1 P27 

分 类 号：S859[农业科学—临床兽医学]