[关键词]
[摘要]
目的 研究尿石素A对乳腺癌细胞MCF-7增殖、凋亡的影响并探讨其作用机制。方法 CCK-8法考察不同浓度尿石素A作用12、24、36、48 h对MCF-7细胞增殖能力的影响;细胞凋亡染色法考察20、40 μmol/L的尿石素A对MCF-7细胞凋亡的影响;实时荧光定量PCR(RT-qPCR)检测c-Myc、Cyclin D1、Bcl-2、Bax mRNA的表达水平;Western blotting法检测c-Myc、Cyclin D1、Bcl-2、Bax蛋白的表达水平。结果 尿石素A对MCF-7细胞增殖具有抑制作用且呈时间浓度相关性;细胞凋亡染色显示,20、40 μmol/L尿石素A给药后均能够诱导MCF-7细胞凋亡;RT-qPCR及Western blotting结果显示,20、40 μmol/L尿石素A能够显著降低MCF-7细胞中c-Myc、Cyclin D1、Bcl-2 mRNA及蛋白的表达水平(P<0.05、0.01),升高Bax mRNA及蛋白的表达水平(P<0.05、0.01)。结论 尿石素A具有抑制MCF-7细胞增殖并诱导其凋亡的作用,其作用机制可能与抑制c-Myc、Cyclin D1、Bcl-2表达,升高Bax表达水平有关。
[Key word]
[Abstract]
Objective To study the effect of urolith A on the proliferation and apoptosis of MCF-7 in breast cancer cells and to explore its mechanism. Methods The CCK-8 cell viability assay was used to investigate the effect of different concentrations of urolith A on MCF-7 proliferation of breast cancer cells after 12, 24, 36 and 48 h. Apoptosis staining was used to investigate the effect of urolith A at 20 and 40 μmol/L on the apoptosis of MCF-7 in adenocarcinoma cells. Real-time fluorescence quantitative PCR (RTqPCR) was used to detect the expression levels of c-myc, Cyclin D1, bcl-2 and Bax mRNA. Western blotting was used to detect the expression levels of c-myc, Cyclin D1, bcl-2, Bax and other proteins. Results The different concentrations of urolith A inhibited the proliferation of MCF-7 in breast cancer cells in A time-concentration dependent manner. 20, 40 μmol/L Urolith A can induce the apoptosis of MCF-7 in adenocarcinoma cells. RT-qPCR and Western blotting results showed that urolith A at 20 and 40 μmol/L could reduce the expression level of c-myc, Cyclin D1, Bcl-2 gene protein and mRNA in MCF-7 cells (P<0.05 and 0.01), and increase the expression level of Bax gene protein and mRNA (P<0.05 and 0.01). Conclusion Urolith A can inhibit the proliferation of MCF-7 in breast cancer cells and induce its apoptosis, and its mechanism may be related to the inhibition of the expression of cmyc, Cyclin D1, Bcl-2 and other genes, and the increase of the expression level of Bax gene.
[中图分类号]
R285.5
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