参芪扶正注射液对人乳腺癌MDA-MB-231细胞增殖和凋亡的影响

目的:研究参芪扶正注射液对基底细胞样(basal-like型)乳腺癌MDA-MB~(-2)31细胞增殖和凋亡的影响,探讨参芪扶正注射液对三阴性乳腺癌的治疗作用机制。方法:体外培养人乳腺癌MDA-MB~(-2)31细胞,将其分为参芪扶正注射液组和空白组。采用噻唑蓝(MTT)比色法分别于24,48,72 h检测参芪扶正注射液对人乳腺癌MDA-MB~(-2)31细胞增殖的影响。采用流式细胞术(FCM)测定细胞周期分布和凋亡情况。实时荧光定量PCR(Real-time PCR)和免疫印迹(Western blot)检测参芪扶正注射液对MDA-MB~(-2)31细胞p53正向细胞凋亡调控因子(PUMA)表达的影响。结果:参芪扶正注射液可抑制MDA-MB~(-2)31细胞增殖,高浓度者抑制作用强,即呈现浓度依赖性,体外培养48 h时抑制作用最显著。参芪扶正注射液将MDA-MB~(-2)31细胞阻滞于细胞周期的S期,进而诱导MDA-MB~(-2)31细胞凋亡。Real-time PCR和Western blot发现参芪扶正注射液可上调MDA-MB~(-2)31细胞PUMA蛋白和基因表达。结论:参芪扶正注射液可显著抑制人乳腺癌MDA-MB~(-2)31细胞增殖,导致细胞周期阻滞,诱导细胞凋亡,其机制可能与上调PUMA基因有关。

Effects of Shenqi Fuzheng Injection on Proliferation and Apoptosis of Human Breast Cancer MDA-MB-231 Cells

Objective: To study the effects of Shenqi Fuzheng injection on proliferation and apoptosis of basal-like type human breast cancer MDA-MB-231 cells, and investigate the treatment mechanism of Shenqi Fuzheng injection for triple negative breast cancer. Method: The human breast cancer MDA-MB-231 cells cultured in vitro were divided into Fuzheng injection groups (six groups with different concentrations of Shenqi Fuzheng injection) and blank group. Methyl thiazolyl tetrazolium (MTT) assay was used to detect the effect of Shenqi Fuzheng injection on proliferation of MDA-MB-231 cells at 24, 48 and 74 h. Flow cytometry (FCM) was used to detect the cell cycle distribution and apoptosis of MDA-MB-231 cells. Real-time PCR and Western blot assay were used to detect the effect of Shenqi Fuzheng injection on p53 up-regulated modulators of apoptosis (PUMA) in MDA-MB-231 cells. Result: Shenqi Fuzheng injection could inhibit the proliferation of MDA-MB-231 cells in a concentration-dependent manner, and the inhibitory effect was most obvious at 48 h in vitro culture. Shenqi Fuzheng injection blocked cell cycle at S phase of the MDA-MB-231 cells, and thus induced apoptosis of MDA-MB-231 cells. Real-time PCR and Western blot assay showed that Shenqi Fuzheng injection could up-regulate PUMA expression of MDA-MB-231 cells. Conclusion: Shenqi Fuzheng injection can significantly inhibit proliferation and block cell cycle at S phase of the MDA-MB-231 cells, thus induce apoptosis of the cells, and the mechanism may be associated with up-regulating PUMA mRNA expression.