曲古霉素A对肺腺癌细胞NCI-H1299增殖抑制作用及机制  

Role for trichostatin A in the proliferation of human lung adenocarcinoma cell NCI-H1299

作　　者：王昌惠[1] 顾红军 武宁[3] 胡海洋[3] 

机构地区：[1]同济大学附属第十人民医院呼吸内科,上海200072 [2]解放军第100医院呼吸内科 [3]第二军医大学附属长海医院呼吸内科,上海200433

出　　处：《同济大学学报（医学版）》2009年第5期10-14,18,共6页Journal of Tongji University(Medical Science)

基　　金：国家自然科学基金资助项目(30670913)

摘　　要：目的观察组蛋白去乙酰化酶(histone deacetylase)抑制剂曲古霉素A(trichostatin A,TSA)对体外培养肺腺癌NCI-H1299细胞生长情况及相关基因表达的影响,并探讨其可能的作用机制。方法MTT法检测不同浓度(0.1、0.2、0.4、2.0μmol/L)的TSA对人肺腺癌NCI-H1299生长的影响,流式细胞术检测处理后NCI-H1299细胞周期分布及凋亡率的变化;Western印迹法检测处理后人肺腺癌NCI-H1299组蛋白H4乙酰化水平的变化;Real-TimePCR检测处理后人肺腺癌NCI-H1299、p21、cyclin B1、Bcl-2和Bax的表达。结果TSA体外能明显抑制NCI-H1299细胞生长,且抑制作用呈明显的剂量、时间依赖性。0.4μmol/L TSA诱导后,流式细胞术检测示细胞阻滞于G2/M期;TSA可明显提高NCI-H1299组蛋白乙酰化水平,并诱导p21和Bax的mRNA表达和抑制Bcl-2和cyclin B1的mRNA表达。结论TSA可通过诱导细胞凋亡及细胞周期阻滞而发挥体外抗肺腺癌细胞株作用,其作用机制可能涉及组蛋白乙酰化水平以及诱导调控相关基因p21、Bax、Bcl-2和cyclin B1。Objective Trichostatin A （TSA）, an antifungal antibiotic with cytostatic and differentiating properties in mammalian cell culture, is a potent and specific inhibitor of histone deacetylase （ HDAC ） activity. This study aims to investigate the influence of trichostatin A on the growth of human lung adenocacinoma cells and on the expression of related genes, and to explore the mechanism involved in vitro. Methods MTr assay was employed to evaluate the inhibitory effect of TSA （0. 1, 0.2, 0.4 and 2.0 gmol/L） on growth of human NCI-H1299 cancer cells. The cell cycle distribution and apoptotic ratio were determined by flow cytometry. The acetyl level of histone after TSA treatment was detected bv Western blot r the mRNA expression of Bax, Bcl-2, p21 and cyclin B1 was measured by Real-Time PCR. Results MTT assay revealed TSA inhibited the growth of NCI-H1299 cells in a concentration- and time-dependent manner. Flow cytometry showed that the cells were blocked at G2/M phase. TSA obviously promoted acetyl level of histone H4, induced expression of p21 and Bax, and inhibited expression of cyclin B1 and Bcl-2. Conclusion TSA can inhibit the proliferation of lung adenocarcinoma cancer cells through inducing cell apoptosis and cell cycle arrest in vitro, which might be related to acetyl level of histone and the expression of p21, Bax, Bcl-2 and cyclinB1.

关 键 词：组蛋白去乙酰化酶 肺肿瘤 曲古霉素A 凋亡 

分 类 号：R734.2[医药卫生—肿瘤]