GADD45β过表达抑制膀胱癌细胞T24的增殖  

Overexpression of GADD45β Inhibited Proliferation of Bladder Cancer T24 Cells

作　　者：宋丽丽[1] 任璐[1] 张婷婷[1] 马恩波[1] 郑耀武[1] 任连生[2] 张建珍[1] 

机构地区：[1]山西大学应用生物学研究所,山西太原030006 [2]山西省肿瘤研究所,山西太原030001

出　　处：《山西大学学报（自然科学版）》2014年第4期625-629,共5页Journal of Shanxi University(Natural Science Edition)

基　　金：山西省实验动物专项(2010K02);山西省国际科技合作项目(2013081051)

摘　　要：GADD45β(growth arrest and DNA damage,GADD)在肿瘤的发生、发展以及凋亡过程中起着重要作用。首先利用Real-time quantitative(qPCR)检测到32μg/mL巴西苏木素处理人类移行膀胱癌T24细胞6h后,GADD45β的mRNA水平显著上调2.7倍。为了研究GADD45β在巴西苏木素致死T24细胞中的作用,从T24细胞的cDNA中扩增获得GADD45β基因全长读码框,并将其克隆到真核表达载体pcDNA3.1上,构成重组载体pcDNA3.1-GADD45β。将该真核表达载体转染T24细胞,发现GADD45β在mRNA水平的表达显著上调,表明真核表达载体成功转入T24细胞;观察GADD45β过表达后对肿瘤细胞的影响,发现细胞变圆,贴壁性变差,活性显著降低85%。本研究表明GADD45β在T24细胞中的过表达能有效地抑制细胞生长,对细胞有较强的致死作用。GADD45β（growth arrest and DNA damage,GADD）plays an important role in tumor occurrence,development and apoptosis.The latest results of our group showed that the expression level of GADD45βwas significantly up-regulated 2.7-fold in T24 cells after 32μg/mL brazilin treatment for 6hby Real-time quantitatine PCR（qPCR）.In order to investigate the role of GADD45βin T24 cells,the eukaryotic expression vector with GADD45βgene was constructed.The ORF of GADD45βwas amplified by PCR from cDNA of T24 cells and then ligated with the vector pcDNA3.1,the recombined vector was named as pcDNA3.1-GADD45β.The vector pcDNA3.1-GADD45βwas transfected into T24 cells with liprofectamine 2000,and then the significant up-expression of GADD45βin mRNA level was detected by qPCR technology.The results suggested that the eukaryotic expression vector entered into T24 cells successfully.Moreover,the cell morphology of T24 cells was changed after the overexpression of GADD45β,including cells shrinkage and rounding,adherent deterioration,and the viability was reduced to 15%.These results showed that the overexpression of GADD45βin T24 cells can inhibit the growth of cells effectively and has a strong lethal effect on cells.

关 键 词：GADD45Β T24细胞 巴西苏木素 

分 类 号：Q78[生物学—分子生物学]