NNMT在肾透明细胞癌的表达及对肾癌细胞侵袭能力的影响

陈伟浩; 宋尔霖; 张鹏; 范阳; 高宇; 倪栋; 逄海港; 艾青; 马鑫; 张旭

NNMT在肾透明细胞癌的表达及对肾癌细胞侵袭能力的影响

- 中国人民解放军总医院泌尿外科(北京, 100853)
基金项目:
国家自然科学基金资助项目（编号30972982）

详细信息

通讯作者: 张旭,E-mail:xzhang@foxmail.com

中图分类号: R737.11

收稿日期: 2012-12-13

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

- Departments of Urology, Chinese People's Liberation Army General Hospital, Beijing, 100853, China

More Information

Corresponding author: ZHANG Xu, E-mail: xzhang@foxmail.com

Received Date: 13 December 2012

摘要

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）；Transwell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。
- 肾细胞癌 /
- 尼克酰胺-N-甲基化酶 /
- 侵袭
Abstract: 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-O, 30 cases of ccRCC surgical specimens 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-O cells in mediation of liposome. The expression level of NNMT in 786-O 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-O, 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-O 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.
- renal cell carcinoma /
- nicotinamide N methytransferase /
- invasion

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参考文献(13)

[1]	GOTTARDO F, LIU C G, FERRACIN M, et al. Micro-RNA profiling in kidney and bladder cancers[J]. Urol Oncol, 2007, 25:387-392.
[2]	Aksoy S, Brandriff B F, Ward A, et al. Human nicotinamide N-methyltransferase gene:molecular cloning, structural characterization and chromosomal localization[J]. Genomics, 1995, 29:555-561.
[3]	Roessler M, Rollinger W, Palme S, et al. Identification of nicotinamide N-methyltransferase as a novel serum tumor marker for colorectal cancer[J]. Clin Cancer Res, 2005, 11:6550-6557.
[4]	Rogers C D, Fukushima N, Sato N, et al. Differentiating pancreatic lesions by microarray and QPCR analysis of pancreatic juice RNAs[J]. Cancer Biol Ther, 2006, 5:1383-1389.
[5]	Emanuelli M, Santarelli A, Sartini D, et al. Nicotinamide N-Methyltransferase upregulation correlates with tumour differentiation in oral squamous cell carcinoma[J]. Histol Histopathol, 2010, 25:15-20.
[6]	Lim B H, Cho B I, Kim Y N, et al. Overexpression of nicotinamide N-methyltransferase in gastric cancer tissues and its potential post-translational modification[J]. Exp Mol Med, 2006, 38:455-465.
[7]	Wu Y, Siadaty M S, Berens M E, et al. Overlapping gene expression profiles of cell migration and tumor invasion in human bladder cancer identify metallothionein 1E and nicotinamide N-methyltransferase as novel regulators of cell migration[J]. Oncogene, 2008, 27:6679-6689.
[8]	Bromberg A, Lerer E, Udawela M, et al. Nicotinamide-N-methyltransferase (NNMT) in schizophrenia:genetic association and decreased frontal cortex mRNA levels[J]. Int J Neuropsychopharmacol, 2012, 15:727-737.
[9]	D'Andrea F P, Safwat A, Kassem M, et al. Cancer stem cell overexpression of nicotinamide N-methyltransferase enhances cellular radiation resistance[J]. Radiother Oncol, 2011, 99:373-378.
[10]	Pozzi V, Mazzotta M, Lo Muzio L, et al. Inhibiting proliferation in KB cancer cells by RNA interference-mediated knockdown of nicotinamide N-methyltransferase expression[J]. Int J Immunopathol Pharmacol, 2011, 24:69-77.
[11]	Sartini D, Pozzi V, Renzi E, et al. Analysis of tissue and salivary nicotinamide N-methyltransferase in oral squamous cell carcinoma:basis for the development of a noninvasive diagnostic test for early-stage disease[J]. Biol Chem, 2012, 393:505-511.
[12]	Riester M, Taylor J M, Feifer A, et al. Combination of a novel gene expression signature with a clinical nomogram improves the prediction of survival in high-risk bladder cancer[J]. Clin Cancer Res, 2012, 18:1323-1333.
[13]	Tomida M, Ohtake H, Yokota T, et al. Stat3 up-regulates expression of nicotinamide N-methyltransferase in human cancer cells[J]. J Cancer Res Clin Oncol, 2008, 134:551-559.