SB939诱导乳腺癌细胞周期阻滞和凋亡的作用及机制  

SB939 efficiently induce breast cell lines cycle arrest and apoptosis and the mechanism

作　　者：李媛媛[1] 张斌[1] 张月[1] 冯炜红[1] 赵洪猛[1] 曹旭晨[1] 

机构地区：[1]天津医科大学附属肿瘤医院乳腺一科乳腺癌防治教育部重点实验室天津市“肿瘤防治”重点实验室,300060

出　　处：《中华实验外科杂志》2012年第5期790-792,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81001186）;天津市自然科学基金资助项目（10JcYBJc14100）

摘　　要：目的观察组蛋白去乙酰化酶（HDAC）抑制剂SB939对乳腺癌细胞株T47D和MDA—MB-231增殖、细胞周期及凋亡的影响。方法不同浓度SB939处理细胞后，用噻唑蓝（MTY）比色法检测对细胞增殖影响；碘化丙锭（PI）染色法检测细胞周期变化；膜联蛋白V（AnnexinV）／PI双染法检测细胞凋亡；Westernblot检测聚腺苷二磷酸核糖聚合酶（PARP）等蛋白表达水平；实时定量聚合酶链反应（ReM-timePCR）检测p21及细胞周期蛋白（Cyclin）B1mRNA水平。结果SB939明显抑制上述两种细胞增殖，24h半数抑制浓度（IC50）分别为（2．721±0．320）、（5．647±0．470）μmoL／L；1μmoL／L药物作用细胞24h，G2／M期比例增加，分别为（21．20±1．90）％、（28．35±2．25）％，凋亡细胞比例增加，分别为（37．60±1．34）％、（34．40±1．77）％，与对照组比较，差异有统计学意义（P〈0．05）；PARP剪切明显增加，bax、p21表达增多，B淋巴细胞／白血病-2（bcl-2）、cdc2、细胞周期蛋白（Cyclin）B1表达减少；p21mRNA水平增加，CyclinB1mRNA水平减少。结论SB939在体外条件下能够明显抑制乳腺癌细胞增殖，诱导细胞周期阻滞及促进细胞凋亡，并呈明显的量效关系。Objective To investigate the effects of SB939, a potent histone deacetylase （HDAC） inhibitor, on proliferation, cycle arrest and apoptosis of breast cancer cell lines T47D and MDA-MB-231 in vitro and its potential regnlating mechanism. Methods Cells were treated with different concentrations of SB939. Cell proliferation was measured by using methyl thiazol tetrazolium （MTT） assay. Cell cycle rate was examined by proliferation index （PI） staining. Apoptotie rate of cells were detected by Annexin V/PI double-staining. The expression of proteins, such as poly ADP-ribose polymerase （PARP）, were analyzed by Western blotting. The mRNA level of p21 and Cyclin B1 were detected by Real-time polymerase chain reaction （Real-time PCR）. Results Along with the increase of SB939 concentration, the inhibition of SB939 on T47D and MDA-MB-231 cell proliferation were obvious enhancement. 50% inhibitory concentration （IC50） were （2. 721±0. 320）, （5. 647±0. 470） μmol/L; Flow eytometry showed cells in GE/M phase significantly increased （ P 〈 0.05 ） . After cells exposed to SB939 （ 1 μmol/L） for 24 h, G2/M phase rates were （ 21.20 ± 1.90 ） % , （ 28. 35± 2.25 ） %. The rates of apoptotie cells were （ 37.60 ± 1.34 ） % , （ 34.40 ± 1.77 ） % , significantly increased as compared to the control cells （ P 〈 0. 05 ） ; Western blotting showed that the expression of cleaved PARP, bax and p21 were increased while B lymphoeytes/ leukemia-2 （bcl-2） , edc2, Cyclin B1 were down-regnlated; The Real-time PCR suggested that the relative mRNA level of p21 was increased and Cyelin B1 was decreased. Conclusion SB939 in vitro can signifi- cantly inhibit proliferation, induce cell cycle arrest and apoptosis on breast cancer cen lines, which was in a dose-dependent manner.

关 键 词：乳腺肿瘤 细胞周期 细胞凋亡 

分 类 号：R737.9[医药卫生—肿瘤]