MCCC2在前列腺癌细胞系中的表达及其对前列腺癌转移影响的初步研究  

The role of MCCC2 gene expression in promoting prostate cancer cells metastasis

作　　者：李科[1] 陈怡[2] 黄文涛[1] 胡成[1] 李茂胤 钟文文[1] 邱剑光[1] 

机构地区：[1]中山大学附属第三医院泌尿外科,广州510630 [2]中山大学孙逸仙纪念医院乳腺外科,广州510120

出　　处：《中华腔镜泌尿外科杂志（电子版）》2014年第2期55-59,共5页Chinese Journal of Endourology(Electronic Edition)

基　　金：广东省科技计划社会发展项目(2010B031600083)

摘　　要：目的分析比较MCCC2基因在不同类型前列腺癌细胞系中的表达差异,通过构建前列腺癌MCCC2基因稳定表达细胞株研究其表达对前列腺癌细胞迁移能力的影响,初步证实MCCC2基因在前列腺癌进展转移中的意义。方法利用RT-PCR方法检测MCCC2基因在局限性和转移性前列腺癌细胞中的表达情况,分析其表达与前列腺癌进展转移的关系;以前列腺癌细胞基因组cDNA为模板,通过PCR扩增MCCC2基因,重组构建含有Flag标签的pCDNA3.1(+)-Flag-MCCC2质粒,包装慢病毒,转染前列腺癌细胞,通过荧光显微镜、RT-PCR和Western Blot等方法鉴定MCCC2基因在细胞内的表达情况;利用Transwell及划痕实验研究MCCC2基因对前列腺癌细胞迁移能力的改变。结果较局限性前列腺癌细胞22Rv1,MCCC2基因mRNA在转移性前列腺癌细胞LNCaP、PC3及DU145中的表达显著升高。同时,激素非依赖性前列腺癌细胞中MCCC2 mRNA表达比激素依赖性前列腺癌细胞明显增强。进而,我们成功构建和包装了表达MCCC2基因的pCDNA3.1(+)-Flag-MCCC2重组质粒及慢病毒,并转染前列腺癌细胞,筛选出稳定表达株;并且,通过Transwell和划痕实验证实MCCC2基因表达与前列腺癌细胞的迁移能力呈正性关联。结论局限性和转移性前列腺癌细胞、激素依赖性和非依赖性前列腺癌细胞中MCCC2基因的表达存在明显差异,提示MCCC2可能在前列腺癌的进展转移过程中发挥重要作用;成功构建了MCCC2基因稳定表达的前列腺癌细胞株,并初步证实其过表达可以促进前列腺癌细胞的迁移能力,为进一步研究MCCC2基因在前列腺癌进展、转移等过程中的具体作用奠定了实验基础。Objective To reveal the differences of MCCC2 gene expression between prostate cancer cell types, and initially proved its expression was associated with prostate cancer cells migration. Methods We compared MCCC2 expression between prostate cancer cell types using RT-PCR. Meanwhile, MCCC2 gene was amplified from prostate cancer cells, and the eukaryotic expression plasmid pCDNA3.1 （＋）-Flag -MCCC2 and lentivirus Lenti-MCCC2 was constructed, respectively. After transfecting MCCC2-plasmid and lentivirus in prostate cancer cells, RT- PCR and Western Blot was performed to detect MCCC2 gene expression, and then Transwell and wound scratch assay were performed to investigate the alteration of migration by overexpressing MCCC2 in prostate cancer cells. Results We demonstrated that was significantly different between prostate cancer cell types in MCCC2 mRNA expression, and MCCC2 was low expressed in localized prostate cancer cells compared to metastasis prostate cancer cells, while MCCC2 with higher expression in hormone- independent prostate cancer cells than androgen-dependent prostate cancer cells. Furthermore, we successfully constructed eukaryotic expression plasmid and lentivirus containing MCCC2 gene. After that, the prostate cancer cell lines which stably overexpressed MCCC2 was established after cells selecting. Finally, we demonstrated that up- regulation of MCCC2 expression has ability to enhance the migration of prostate cancer cells. Conclusion Our study provides a potential target gene MCCC2 that may play the antagonistic role in tumor development, and establishe the foundations for the further research in MCCC2 function in prostate cancer.

关 键 词：前列腺癌 MCCC2 雄激素受体 

分 类 号：R737.25[医药卫生—肿瘤]