生长抑制因子4对人黑素瘤细胞株M14凋亡的调控  

The Regulatory Effect of ING4 on Apoptosis in Melanoma Cell Lines M14

作　　者：蔡丽敏[1,2] 李海燕 孙建方 

机构地区：[1]中国医学科学院皮肤病研究所 [2]中国协和医科大学皮肤病医院病理科,江苏南京210042 [3]黑龙江省肿瘤医院皮肤科,黑龙江哈尔滨150076

出　　处：《中国皮肤性病学杂志》2008年第10期577-580,共4页The Chinese Journal of Dermatovenereology

摘　　要：目的探讨生长抑制因子4对人黑素瘤细胞株M14凋亡的调控及其作用机制。方法构建人生长抑制因子4基因的真核表达质粒,并将其转导入M14细胞,采用流式细胞术及TUNEL法检测细胞的凋亡情况,同时采用蛋白印迹法分析凋亡通路的活化及通路相关蛋白的表达情况。结果转染外源生长抑制因子4基因后,M14细胞出现了明显的凋亡峰并伴有G2/M期阻滞,凋亡率及凋亡指数也明显高于转染空载质粒及未转染M14细胞,差异有显著性意义(P<0.01)。还发现转染外源生长抑制因子4基因后,M14细胞凋亡通路活化,抗凋亡蛋白Bcl-2表达下调,而促凋亡蛋白Bax表达上调,凋亡执行蛋白Cyt-c表达增加而caspase3表达下降。结论生长抑制因子4可能通过活化凋亡通路,进而促进M14细胞凋亡。这可能也是ING4抑制M14细胞增殖的机制之一。Objective To investigate the regulatory effect of ING4 on apoptosis and its mechanism in melanoma cell lines M14. Methods pCDNA3.1-ING4 eukaryotic expression vector was constructed and then transfected into Mid cells. Cell cycle and apoptotic percentage were detected by flow cytometry, apoptotic cells were quantitatively determined by TUNEL method. In addition, the expression of apoptosis pathway-related proteins were examined by western blot analysis. Results Mid clones with exogenous ING4 gene presented with the enhanced apoptotic peak and percentage accompanied with the enhanced G2/M arrest, as well apoptotic cells were found more than those in non-transected Mid clones and M14 clones with empty plasmid （P 〈 0.01 ）. And moreover, apoptosis pathway was activated in M14 clones with exogenous ING4 gene, and the downregulation of Bcl-2 and caspase3 and the upregulation of Bax and Cyt-c（ cytochrome C） were found. Conclusion ING4 might induce M14 cells apoptosis by activating apoptosis pathway via the change of apoptosis-related proteins expression. It might be one of anticancer mechanisms of ING4.

关 键 词：生长抑制因子4 黑素瘤 凋亡 

分 类 号：R739.5[医药卫生—肿瘤]