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p53基因是一种重要的抑癌基因,其产物能够与靶DNA特异性结合并激活转录参与诱导细胞凋亡,调控细胞周期,控制细胞的增殖与分化.p44蛋白主要定位于前列腺癌细胞的细胞质,促进癌细胞的增殖.通过shRNA沉默p44的表达后,LNCaP细胞的生长受抑制,p53靶基因TIGAR和GLIPR1的表达增加.为探寻p53是否介导p44对TIGAR和GLIPR1的作用,本研究构建了pGL3-4×PRE-E4-luc报告基因质粒,并将其分别转染经NT-shRNA慢病毒和p44-shRNA慢病毒感染的LNCaP细胞,比较两者荧光素酶活性.结果显示,重组质粒pGL3-4×PRE-E4-luc构建成功,为进一步研究p53在肿瘤发生发展过程中的作用通路提供了新的手段.但LNCaP细胞中p44并非通过p53作用于TIGAR和GLIPR1基因,具体机制仍有待进一步研究.
The p53 gene is an important tumor suppressor gene whose product can specifically bind to target DNA and activate transcription to participate in the induction of apoptosis, regulate the cell cycle, and control the proliferation and differentiation of cells. The p44 protein is mainly located in the cytoplasm of prostate cancer cells , And promote the proliferation of cancer cells.After silencing the expression of p44 by shRNA, the growth of LNCaP cells was inhibited and the expression of p53 target genes TIGAR and GLIPR1 were increased.In order to find out whether p53 mediated the effect of p44 on TIGAR and GLIPR1, we constructed pGL3-4 × PRE-E4-luc reporter gene were transfected into LNCaP cells transfected with the lentivirus of NT-shRNA and lentivirus of p44-shRNA respectively.The results showed that the recombinant plasmid pGL3-4 -4 × PRE-E4-luc was successfully constructed, which provided a new method for further study on the role of p53 in tumorigenesis and progression.However, p44 does not act on TIGAR and GLIPR1 through p53 in LNCaP cells, the study.