TSA诱导MCF-7细胞毒性过程中转录调节的实验研究
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南京医科大学创新基金资助项目(CX2002003)


A study on transcription regulation induced by trichostatin A during cytotoxicity on MCF-7 cells
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    摘要:

    目的:研究制滴菌素(trichostatin A,TSA)诱导乳腺癌细胞株MCF-7的细胞毒性作用及基于该效应可能的转录调节基础。方法:采用MTT法观察不同浓度TSA对MCF-7细胞的抑制作用,Annexin-V/PI双染观察该梯度下的细胞凋亡情况,细胞周期分析检测不同时间段的周期变化,RT-PCR分析TSA处理前后ERα、myc-c、P21、cyclin-D及Bcl-2基因的转录表达情况。结果:TSA对MCF-7细胞的抑制作用具有时间和剂量的依赖性,Annexin-V/PI双染分析显示在TSA处理48 h后,随其浓度的增加,细胞的凋亡率依次升高,0.5 -滋mol/L TSA作用,细胞凋亡率为33.82%;细胞周期分析检测显示在0.5 -滋mol/L TSA的作用下,MCF-7细胞出现周期阻滞,但未检测出细胞凋亡。RT-PCR分析显示除P21基因上调外, ERα、myc-c、cyclin-D及Bcl-2均下调,同朝着增殖抑制、周期阻滞及凋亡的方向进展。结论:TSA能诱导MCF-7细胞的细胞毒性作用,其机制可能与转录调节有关。

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    Objective:To investigate cytotoxicity of breast cancer cell line MCF-7 cells induced by trichostatin A and its possible transcription regulation. Methods:The proliferative activity of MCF-7 cells upon different trichostatin A concentration was accessed by MTT assay, cell apoptosis followed was identified by Annexin-V/PI double staining; cycle alteration was examined by cell cycle analysis in different time, expression status of ERα, myc-c, P21, cyclin-D and Bcl-2 genes was analyzed by RT-PCR. Results:A time and dose dependent inhibition was detected in MCF-7 cells treated with trichostain A. At 48 h treatment, Annexin-V/PI double staining analysis indicated that cell apoptosis rate increased as trichostatin A rose, at 0.5 -滋mol/L treatment, apoptosis rate was 33.82%; Cell cycle analysis showed MCF-7 cells were arrested in cycles after trichostatin A treatment, while almost no apoptosis was evidenced. RT-PCR revealed that except P21 gene,ERα,myc-c,cyclin-D and Bcl-2 were all down regulated,and all made it to proliferative inhibition, cell arrest and apoptosis. Conclusion:Trichostatin A could induce cytotoxicity in MCF-7 cells, and may correlate with transcription regulation.

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朱义保,潘 梅,魏钦俊,曹新.TSA诱导MCF-7细胞毒性过程中转录调节的实验研究[J].南京医科大学学报(自然科学版),2007,(6):546-549

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  • 收稿日期:2006-10-20
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