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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 renal clear cell carcinoma and its influence on the invasiveness of renal cell carcinoma. Methods: RT-PCR and Western blot were used to detect the expression of NNMT mRNA and protein in normal renal tubular epithelial cell line HKC, renal carcinoma cell line 786-O, and renal clear cell carcinoma tissues of 30 cases. The relationship between NNMT mRNA level and clinicopathological parameters was analyzed. The NNMT-specific siRNA sequences were chemically synthesized and transfected into 786-O cells using lipofectamine 2000. The expression of NNMT in 786-O cells was detected by RT-PCR and Western blot. Detection of renal cell carcinoma 786-O invasion changes. Results: The mRNA and protein expression of NNMT in renal cell carcinoma 786-O was significantly higher than that in normal renal tubular epithelial cell line HKC (P <0.001); relative expression of NNMT mRNA in renal clear cell carcinoma tissues and corresponding paracancerous tissues (1.582 ± 0.2145) and (0.1269 ± 0.04279) respectively, P <0.001 in both groups. The mRNA level of NNMT correlated with tumor size and clinical stage (P <0.05). Tran-swell assay showed that the invasion ability of 786-O cells was significantly decreased after NNMT was decreased. Conclusion: The expression of NNMT in renal clear cell carcinoma tissues and cells may play an important role in the occurrence and development of renal cell carcinoma.