| 温海燕,李 娜,侯亚婷,戴 谆,宋金春.甲基莲心碱抗乳腺癌 MCF-7细胞的研究[J].,2018,(18):3425-3430 |
| 甲基莲心碱抗乳腺癌 MCF-7细胞的研究 |
| Study on the Effects of Neferine on MCF-7 Cells |
| 投稿时间：2018-04-13 修订日期：2018-05-10 |
| DOI：10.13241/j.cnki.pmb.2018.18.005 |
| 中文关键词: 甲基莲心碱 乳腺癌 增殖 凋亡 |
| 英文关键词: Neferine Breast cancer Proliferation Apoptosis |
| 基金项目:武汉市临空港经济开发区 "金山英才计划 "课题基金项目 |
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| 中文摘要: |
| 摘要 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。 |
| 英文摘要: |
| 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. |
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