【摘 要】
：
目的 研究曲古抑素A (Trichostatin A,TSA)诱导肺癌细胞株A549细胞周期阻滞及细胞凋亡及其相互作用,并探讨其可能的作用机制.方法 采用MTT法检测不同浓度TSA对A549细胞的抑制作用,流式细胞仪检测不同浓度及时间段细胞凋亡的情况及细胞周期的变化.Western印迹检测处理后A549细胞H3乙酰化表达水平.加入Caspase抑制剂Z-VAD-FMK后检测TSA处理细胞的凋亡及周
【机 构】
：
武汉华中科技大学附属同济医院生物医学中心实验室 430030
【出 处】
：
中华医学会第十四次全国妇科肿瘤学术会议
论文部分内容阅读
目的 研究曲古抑素A (Trichostatin A,TSA)诱导肺癌细胞株A549细胞周期阻滞及细胞凋亡及其相互作用,并探讨其可能的作用机制.方法 采用MTT法检测不同浓度TSA对A549细胞的抑制作用,流式细胞仪检测不同浓度及时间段细胞凋亡的情况及细胞周期的变化.Western印迹检测处理后A549细胞H3乙酰化表达水平.加入Caspase抑制剂Z-VAD-FMK后检测TSA处理细胞的凋亡及周期变化.结果 TSA对A549细胞的抑制作用具有时间和剂量的依赖性,0.5umol/L TSA诱导12-24h内,流式细胞术检测示细胞阻滞于G2/M期,随着时间延长至36-48h,G2/M期阻滞的细胞趋向凋亡.TSA可明显提高A549细胞H3乙酰化水平.结论 TSA能诱导A549细胞G2/M期阻滞及凋亡,Caspase抑制剂ZVAD-FMK可逆转TSA诱导的细胞凋亡,但未能逆转G2/M期阻滞.因此TSA诱导G2/M期阻滞后激活凋亡信号通路,从而诱导细胞凋亡.
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目的 现在已经有很多相关研究证实Fezl是一种抑癌基因,在多种肿瘤组织中表达较正常组织来说有不同程度的降低.在肿瘤细胞中将Fezl进行过表达之后能够使肿瘤细胞停留在G2/M期的细胞增加.本实验旨在研究Fezl基因在调控乳腺癌细胞系MCF-7生长与增殖中的作用.方法 构建Fezl-GFP质粒并将其与GFP质粒分别转染入MCF-7细胞中,通过流式细胞仪检测MCF-7细胞周期的变化.