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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。
Objective: To investigate the effect of Shenqi Fuzheng injection on the proliferation and apoptosis of basal-like breast cancer MDA-MB -2-231 cells and to explore the therapeutic effect of Shenqi Fuzheng injection on triple negative breast cancer Mechanism. Methods: Human breast cancer MDA-MB ~ (-2) 31 cells were cultured in vitro and divided into Shenqi Fuzheng injection group and blank control group. The effects of Shenqi Fuzheng Injection on the proliferation of human breast cancer MDA-MB ~ (-2) 31 cells were detected by MTT colorimetry at 24, 48 and 72 h respectively. Cell cycle distribution and apoptosis were determined by flow cytometry (FCM). The effects of Shenqi Fuzheng Injection on the expression of p53 positive apoptotic regulatory factor (PUMA) in MDA-MB -2-231 cells were detected by Real-time PCR and Western blot. Results: Shenqi Fuzheng injection could inhibit the proliferation of MDA-MB-2-31 cells in a concentration-dependent manner. The inhibitory effect was the most significant at 48 hours. Shenqi Fuzheng injection blocked MDA-MB ~ (-2) 31 cells in the S phase of the cell cycle, and then induced apoptosis of MDA-MB ~ (-2) 31 cells. Real-time PCR and Western blot showed that Shenqi Fuzheng injection could up-regulate the PUMA protein and gene expression in MDA-MB-21 cells. Conclusion: Shenqi Fuzheng injection can significantly inhibit the proliferation of human breast cancer MDA-MB ~ (-2) 31 cells, lead to cell cycle arrest and induce apoptosis, and its mechanism may be related to the up-regulation of PUMA gene.