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作 者:陈伟浩[1] 宋尔霖[1] 张鹏[1] 范阳[1] 高宇[1] 倪栋[1] 逄海港 艾青[1] 马鑫[1] 张旭[1]
机构地区:[1]中国人民解放军总医院泌尿外科,北京100853
出 处:《临床泌尿外科杂志》2013年第4期241-245,共5页Journal of Clinical Urology
基 金:国家自然科学基金资助项目(编号30972982)
摘 要:目的:研究NNMT在肾透明细胞癌中的表达情况及对肾癌细胞侵袭能力的影响。方法:采用RT-PCR和Western blot方法检测正常肾小管上皮细胞株HKC、肾癌细胞株786-O及30例肾透明细胞癌组织、相应癌旁组织中NNMT的mRNA和蛋白的表达水平,并分析NNMT的mRNA水平与临床病理参数的关系。化学合成针对NNMT特异的siRNA序列,应用脂质体Lipofectamine 2000将其转染进786-O细胞中,利用RT-PCR和Western blot法检测NNMT在786-O细胞中的表达水平,用Transwell小室法检测肾癌细胞786-O侵袭能力的变化。结果:NNMT在肾癌细胞786-O中的mRNA和蛋白表达水平显著高于正常肾小管上皮细胞株HKC(P<0.001);肾透明细胞癌组织和对应的癌旁组织中NNMT的mRNA相对表达量分别为(1.582±0.2145)、(0.1269±0.04279),两组比较P<0.001。NNMT的mRNA水平与肿瘤大小、临床分期有关(P<0.05);Tran-swell法检测结果显示降低NNMT的表达后786-O细胞的侵袭能力明显下降。结论:NNMT在肾透明细胞癌组织和细胞中表达升高,可能在肾癌发生、发展过程中发挥重要作用。Objective:To study the expression of NNMT in clear-cell renal cell carcinoma (ccRCC) and its role in renal cell carcinoma invasion in vitro. Method: The expression of NNMT in mRNA and protein levels were detected in human.kidney tubule epithelial cell line HKC, renal cell cancer line 786-0, 30 cases of ccRCC surgical speci mens and their corresponding adjacent tissues, using real-time PCR and Western blot respectively. And the relationship between the mRNA levels of NNMT and clinic-pathological parameters were analyzed . The special siRNA sequence of NNMT was synthesized chemically, and transfected into 786-0 cells in mediation of liposome. The expression level of NNMT in 786-0 cells was determined by RT-PCR and Western blot; and the changes in cell invasion were detect by Transwell assays. Result: The mRNA and protein levels of NNMT were significantly up regulated in ccRCC cell lines of 786-0, compared to human kidney tubule epithelial cell line HKC ( P 〈0. 001). Meanwhile, the mRNA levels of NNMT in ccRCC surgical specimens and adjacent normal renal tissues were (1. 582±0. 2145), (0. 1269+0. 04279)( P 〈0. 001). The mRNA levels of NNMT in renal carcinoma were significantly related with some clinic-pathologic parameters, such as tumor size and clinicopathological stage ( P〈0.05). The results of Transwell assay indicated that down-regulated the expression of NNMT significantly inhibited the invasion ability of 786-0 cells. Conclusion: NNMT was up-expressed in ccRcc tissues and renal cancer cells, and it may be related to the carcinogenesis and progression of renal cell carcinoma.
关 键 词:肾细胞癌 尼克酰胺一N一甲基化酶 侵袭
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