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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 study the effects of metformin on the proliferation and cell cycle of mammary tumor cells in dogs, the models of CHMm and CHMp in canine mammary tumors were used as the model, and the cells were treated with 0, 5, 10, 20 and 40 mmol / L metformin. CCK-8 The cell proliferation activity was detected by flow cytometry. The proportion of cell cycle distribution was detected by flow cytometry. The expressions of cyclin D1 and p27 were detected by Western blot. Compared with the control group, the survival rate of CHMm cells in each group was significantly decreased (P <0.05), while CHMp cells in addition to the 5 mmol / L group (P> 0.05) 48 hours after metformin treatment, the survival rate of other groups Significantly lower than the control group (P <0.05), and showed a certain concentration-dependent manner. The proportion of cells in G0 / G1 phase of CHMm increased from (38.7 ± 5.89)% to (70.36 ± 5.78)% by flow cytometry, and the percentage of cells in G0 / G1 phase of CHMm increased from (37.03 ± 4.23)% Increase to (54.92 ± 4.67)%. With the increase of concentration of metformin, the expression of cyclin D1 in CHMm and CHMp cells decreased, while the expression of p27 showed an upward trend. The results showed that metformin can inhibit the proliferation of mammary tumor cells in vitro, induce G0 / G1 phase arrest, down-regulate cyclin D1 and up-regulate the expression of p27 cell cycle-related proteins, thus providing a theoretical basis for the treatment of breast cancer in dogs .