HO-1超表达载体的构建及其对膀胱癌T24细胞的影响  

Construction and Expression of Eukaryotic Expression Vector HO-1 and its Roles in Bladder Cancer T24 Cells

作　　者：任璐[1] 宋丽丽[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）》2015年第1期150-154,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：血红素氧合酶1(heme oxygenase-1,HO-1)的表达与肿瘤的生长、凋亡和增殖有密切关系。课题组前期的数字基因表达谱结果显示,巴西苏木素处理膀胱癌T24细胞后,HO-1表达显著提高,提示其可能在这一过程中发挥重要作用。为进一步研究HO-1对T24细胞的影响及其是否为巴西苏木素作用于T24细胞的靶标基因,利用Real-Time quantitative PCR(qPCR)验证巴西苏木素处理T24细胞后HO-1基因表达量的变化,克隆HO-1基因,构建真核表达载体pcDNA3.1-HO-1,将该载体转染T24细胞,MTT法检测HO-1超表达后T24细胞的活性变化。结果显示,HO-1在巴西苏木素处理T24细胞12h后显著上调2.2倍;HO-1成功转入T24细胞中后,mRNA表达显著提高93.3%,细胞活性略有升高。上述结果表明HO-1的高表达对T24细胞有保护作用,HO-1不是巴西苏木素作用于T24细胞的靶标分子,其在巴西苏木素处理T24细胞时表达升高可能是由于药物作用后,细胞为了逃避凋亡而产生应激反应所引起的。Heme oxygenase-1（HO-1）is closely associated with the tumor growth,apoptosis and proliferation.The digital gene expression profile showed that the expression of HO-1 changed significantly after brazilin treatment of T24 bladder cancer cell.It is suggested that HO-1 may play an important role in this process.Based on this hypothesis,we focused on the effect of HO-1 on T24 cells and whether HO-1is the target gene for brazilin inhibiting the growth of T24 cells.We used real-time quantitative PCR to validate how HO-1 expression changed,we cloned HO-1 gene and constructed eukaryotic expression vector pcDNA3.1-HO-1,and the vector was transfected into T24 cells to investigate how T24 cell changed after overexpression of HO-1.The results showed that HO-1 was significantly up-regulated in T24 cells after brazilin treatment for 12 h.The mRNA expression level of HO-1significantly increased by 93.3% and cell activity increased slightly after transferred HO-1 into T24 cells.HO-1 has partial protective effect on T24 cell growth and it is not the target gene in the process of brazilin inhibit the T24 cells.The increased expression of HO-1in brazilin treatment may be due to the stress response which produced by the cells avoiding the apoptosis.

关 键 词：HO-1 T24细胞 巴西苏木素 

分 类 号：Q78[生物学—分子生物学]