抑癌基因WWOX对Lewis肺癌细胞c-jun蛋白表达及其转录活性的影响  

Effect of Cancer Suppressor Gene WWOX on Expression and Transcription Activity of c-jun Protein in Lung Cancer Lewis Cells

作　　者：杨巍[1] 王晓明[1] 

机构地区：[1]台州学院医学院生物化学与分子生物学检验教研室,浙江台州318000

出　　处：《中国生物制品学杂志》2009年第9期861-864,共4页Chinese Journal of Biologicals

摘　　要：目的研究抑癌基因WWOX对Lewis肺癌细胞c-jun蛋白表达及其转录活性的影响,探讨WWOX基因的抑癌机制。方法采用脂质体转染法将WWOX基因重组真核表达质粒转染Lewis肺癌细胞,RT-PCR和Western blot法检测WWOX基因mRNA的转录水平和蛋白的表达水平;免疫组化法检测WWOX基因转染后Lewis细胞中c-jun蛋白的表达水平;半定量RT-PCR法检测c-jun调控的4种肿瘤相关基因p21、cyclinD1、FasL及VEGF mRNA的转录水平。结果重组真核表达质粒pcDNA4.0/Myc-His-WWOX转染Lewis细胞后,WWOX基因在mRNA和蛋白水平上均得到表达;与未转染细胞和空载体转染细胞相比,WWOX基因转染细胞胞浆中c-jun蛋白的表达量升高,而细胞核中c-jun蛋白的表达量未见明显差异;p21基因mRNA的转录水平升高,cyclinD1、FasL和VEGF基因mRNA的转录水平降低。结论WWOX基因可在Lewis细胞中表达,其转染Lewis肺癌细胞后,不直接调控c-jun蛋白的表达量,但可影响其转录活性。Objective To investigate the effect of cancer suppressor gene WWOX on the expression and transcription activity of c-jun protein in lung cancer Lewis cells as well as the cancer suppression mechanism of WWOX gene. Methods The constructed eukaryotic expression vector pcDNA4. 0 / Myc-His-WWOX was transfected to Lewis cells in mediation of liposome. The transcription level of WWOX mRNA was determined by RT-PCR, and the expression level of WWOX protein by Western blot. The expression of c- jun protein in transfected Lewis cells was determined by immunohistochemical assay, and the mRNA transcription levels of 4 kinds of tumor-related genes regulated by c-jun, i.e. p21, cyclinD1, FasL and VEGF, by semi-quantitative RT-PCR method. Results WWOX gene was expressed at both mRNA and protein levels in Lewis cells transfected with pcDNA4. 0/Myc-His-WWOX. Compared with those untransfected and transfected with empty vector, the expression level of c-jun protein increased in cytoplasm of Lewis cells transfected with pcDNA4.0 / Myc-His-WWOX, while showed no significant difference in nucleus. The transcription levels of p21 mRNA in Lewis cells transfected with pcDNA4.0 / Myc-His-WWOX increased, while those of cyelinDl, FasL and VEGF mRNAs decreased. Conclusion WWOX gene was expressed in Lewis cells. The transfection of Lewis cells with WWOX gene could not regulate the expression directly, while showed significant effect on transcription activity of c-jun protein.

关 键 词：抑癌基因 WWOX C-JUN 蛋白表达 转录活性 

分 类 号：R73-36[医药卫生—肿瘤]