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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 (GADD) plays an important role in tumorigenesis, development and apoptosis. First, the mRNA level of GADD45β was up-regulated by 2.7-fold after treatment with 32μg / mL Brazilian hematoxylin on human bladder transitional cell carcinoma T24 cells by Real-time quantitative (qPCR) for 6h. In order to study the role of GADD45β in T24 cells induced by hematoxylin in Brazil, the full-length reading frame of GADD45β gene was amplified from cDNA of T24 cells and cloned into the eukaryotic expression vector pcDNA3.1 to construct the recombinant vector pcDNA3.1 -GADD45β. The eukaryotic expression vector was transfected into T24 cells and found that the expression of GADD45β at mRNA level was significantly upregulated, indicating that the eukaryotic expression vector was successfully transfected into T24 cells. The effect of GADD45β overexpression on tumor cells was observed and found to be round and adherent Sexually degraded, activity was significantly reduced by 85%. This study shows that GADD45β overexpression in T24 cells can effectively inhibit cell growth, the cell has a strong lethal effect.