甲基莲心碱抗乳腺癌 MCF-7细胞的研究

目的:观察甲基莲心碱对乳腺癌细胞系MCF-7增殖和凋亡的影响,并探讨其诱导乳腺癌细胞系MCF-7凋亡的可能作用机制。方法:采用体外培养人乳腺癌细胞系MCF-7,CCK-8实验检测不同浓度甲基莲心碱对MCF-7细胞增殖抑制作用;乳酸脱氢酶(LDH)试剂盒(微板法)检测细胞上清液LDH含量;流式细胞术分析甲基莲心碱对MCF-7细胞周期及凋亡的影响;实时定量PCR(RT-PCR)检测线粒体凋亡相关基因Bax和Bcl-2的表达水平。结果:CCK-8、LDH结果显示甲基莲心碱以时间、浓度依耐性的方式抑制乳腺癌MCF-7细胞的增殖及促进细胞毒性的增加;流式细胞术结果表明不同甲基莲心碱作用下MCF-7的平均凋亡率分别为(15.44±0.52)、(18.81±2.24)、(24.26±2.84)、(36.90±3.15)、(59.27±5.86),且使其周期阻滞于G0/G1期;RT-PCR检测结果证明甲基莲心碱可上调乳腺癌细胞中促凋亡基因Bax的表达,而下调抑制凋亡基因Bcl-2。结论:甲基莲心碱以时间和浓度依赖的方式抑制乳腺癌细胞增殖、细胞毒性增加,导致细胞周期于G0/G1阻滞并促进癌细胞凋亡。甲基莲心碱抗乳腺癌的可能作用机制是激活线粒体凋亡途径。

Study on the Effects of Neferine on MCF-7 Cells

ABSTRACT Objective: To investigate the effect of neferine on the proliferation and apoptosis of breast cancer cell line MCF-7, and to explore its possible mechanism. Methods:The human breast cancer cell line MCF-7 were cultured in vitro. CCK-8 assay was used to detect the the anti-proliferation effect of different concentrations of neferine on MCF-7 cells. The supernatant of the cells was assayed for lactate dehydrogenase content by Lactate dehydrogenase (LDH) kit (microplate assay). The effect of neferine on cancer cell cycle distribution and apoptosis was analyzed by flow cytometry. The expression levels of mitochondrial apoptosis-related genes Bax and Bcl-2 were detected by Real-time PCR (RT-PCR). Results:CCK-8 and LDH assay showed that neferine inhibited the proliferation and increased the cytotoxicity of breast cancer cells in a time- and concentration-dependent manner. Flow cytometry showed that the average apoptotic rates of MCF-7 under different concentrations of neferine were (15.44± 0.52), (18.81± 2.24), (24.26± 2.84), (36.90± 3.15), (59.27± 5.86), and its cell cycle arrested at G₀/G₁ phase. PCR detection indicated that the expression of pro-apoptotic protein Bax was increased, and anti-apoptotic protein Bcl-2 was decreased in MCF-7 cells when treated with different concentrations of neferine. Conclusion: Neferine inhibited the proliferation and increased the cytotoxicity of breast cancer cells in a time- and concentration-dependent manner, leading to the cell cycle arresting at G₀/G₁1 and promoting apoptosis of cancer cells. The activation of the mitochondrial apoptotic pathway may be the anti-cancer mechanism of neferine.