苯乙双胍对上皮-间质转化调控生长的抑制作用

Inhibitory effects of phenformin on the epithelial mesenchymal transformation regulation of growth

目的研究苯乙双胍对乳腺癌细胞通过靶向胰岛素样生长因子1受体(IGF1R)途径上皮-间质转化(EMT)调控生长的抑制作用。方法将人乳腺癌细胞株T47D分为3组:实验1组、实验2组与对照组。实验1组与实验2组向培养液中分别加入苯乙双胍1,10μmol·L^(-1),对照组加入等量的磷酸盐缓冲液,测定细胞存活、增殖与凋亡情况;用免疫印迹法检测磷酸化蛋白激酶B(p AKT)、蛋白激酶B(AKT)蛋白表达情况。结果实验1组、实验2组与对照组的人乳腺癌细胞株T47D的细胞存活率分别为(0.49±0.04)%,(0.27±0.05)%,(0.98±0.02)%;这3组的细胞生长速度分别为(1.37±0.06)%,(1.13±0.02)%,(1.93±0.04)%;这3组的凋亡率分别为(31.22±8.24)%,51.94±9.42)%,(8.11±4.22)%;以上3个指标:2个实验组与对照组比较,差异均有统计学意义(均P<0.05);同时,实验2组与实验1组比较,差异也有统计学意义(P<0.01)。实验1组、实验2组的p AKT蛋白相对表达量高于对照组,差异均有统计学意义(均P<0.05);而实验1组、实验2组的AKT蛋白相对表达量低于对照组,差异均有统计学意义(均P<0.05);同时,实验2组与实验1组比较,差异也有统计学意义(P<0.01)。结论苯乙双胍能抑制乳腺癌细胞的生长与增殖,促使细胞凋亡,激活AKT信号途径,可通过靶向IGF1R途径发挥EMT调控生长的作用。

Objective To investigate the inhibitory effects of target insulin-like growth factor 1 receptor（ IGF1 R） pathway of epithelial mesenchymal transition（ EMT） of phenformin in the breast cancer cell growth.Methods The human breast cancer cell line T47 D were divided into three groups： experimental^（-1）,experimental-2 groups and control group. The experimental^（-1）,experimental-2 groups were added to the culture of 1,10 μmol ﹒ L^（-1） phenformin in liquid,respectively; the control group was treated with the same dose of phosphate buffered saline（ PBS）.The cell survival,proliferation and apoptosis were determined. The expression of P-proteinkinase B（ p AKT） and proteinkinase B（ AKT） protein levels were detected by Western blot. Results The cell survival rates in the experimental^（-1）,experimental-2 groups and control group were（ 0. 49 ± 0. 04） %,（ 0. 27 ± 0. 05） %,（ 0. 98 ± 0. 02） %,respectively; the growth rates in the 3 groups were（ 1. 37 ± 0. 06） %,（ 1. 13 ± 0. 02） %,（ 1. 93 ± 0. 04） %,respectively; the rate of apoptosis in the 3 groups were（ 31. 22 ± 8. 24） %,51. 94 ± 9. 42） %,（ 8. 11 ± 4. 22） %,respectively; comparison between experimental^（-1）,experimental-2 groups and control group,the differnce of the three factors had significantly（ all P 〈0. 05）; comparison between experimental^（-1） group and experimental-2 group,the differnce of the three factors had significantly（ all P〈 0. 01）. The relative expression of p AKT protein in the experimental^（-1）,experimental-2 groups were higher than that of the control group（ P 〈0. 05）; while the relative expression of AKT protein were lower than that in control group（ P 〈0. 05）; and the difference between experimental^（-1） group and experimental-2 group was also statistically significant（ all P 〈0. 05）. Conclusion The phenformin can inhibit breast cancer cells growth and proliferation,induce apoptosis,activation of AKT signaling pathway,and it can play a role in regulating the EMT growth by targeting the IGF1 R.