NNMT在肾透明细胞癌的表达及对肾癌细胞侵袭能力的影响

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。

Expression of NNMT in clear-cell renal cell carcinoma and its role in renal cell carcinoma invasion

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.