肾细胞癌中WNK2基因甲基化状态与mRNA表达的研究

WNK2 gene methylation state and mRNA expression in renal cell carcinoma

目的探讨WNK2基因在肾细胞癌(RCC)发生及发展中的作用。方法选取2016年5月至2017年6月本院的肾肿瘤患者58例,分别应用甲基化特异性PCR(MSP)、逆转录-聚合酶链反应(RT-PCR)法检测其RCC及相应癌旁组织、正常组织标本中WNK2基因甲基化状态和mRNA表达情况。结果在58例RCC组织、癌旁组织及正常组织中的WHK2甲基化阳性率分别为65.52%(38/58),34.48%(20/58),10.34%(6/58)。RCC组织中WNK2基因甲基化阳性率明显高于癌旁组织及正常组织。而癌旁组织及正常组织中WNK2基因甲基化发生率比较,差异无统计学意义(P>0.05)。在RCC组织中WNK2基因启动子区甲基化的发生率在患者的不同年龄、性别、肿瘤大小、临床分期、组织学分级等临床资料间比较,差异均无统计学意义(P>0.05);WNK2 mRNA在RCC和癌旁组织、正常组织中的相对表达量分别为(0.54±0.03)、(0.66±0.02)、(0.68±0.02),RCC组织中WNK2 mRNA相对表达量明显低于癌旁组织和正常组织,差异有统计学意义(P<0.05),但癌旁组织及正常组织中WNK2 mRNA相对表达量比较,差异无统计学意义(P>0.05)。在RCC组织中WNK2基因mRNA相对表达量在患者的不同年龄、性别、肿瘤大小、临床分期、组织学分级等临床资料间比较,差异均无统计学意义(P>0.05);甲基化阳性组中WNK2基因的mRNA相对表达量显著低于甲基化阴性组,差异有统计学意义(P<0.05)。结论WNK2基因启动子区甲基化及其mRNA相对表达量降低可能参与了RCC的发生,但与RCC的临床进展无关。WNK2基因启动子区发生甲基化可能是引起其mRNA相对表达降低的原因之一。

Objective To explore the role of the WNK2 gene in the occurrence and development of renal cell carcinoma(RCC).Methods A total of 58 patients with renal tumor in our hospital from May 2016 to June 2017 were selected.Methylation specific PCR(MSP)and reverse tran-scriptase-PCR(RT-PCR)methods were respectively performed to assay the methylation status and mRNA expression of WNK2 in RCC tissues,corresponding respective tumor-adjacent tissues and normal kidney tissues.Results The relative expression of WNK2 mRNA in RCC,paraneoplastic and normal tissues were(0.54±0.03),(0.66±0.02)and(0.68±0.02),respectively,and the relative expression of WNK2 mRNA in RCC tissues was significantly lower than that in paraneoplastic and normal tissues,and the difference was statistically significant(P<0.05),but the comparison between paraneoplastic and normal tissues.The differences were not statistically significant(P>0.05).The relative expression of WNK2 mRNA in RCC tissues was not statistically significant(P>0.05)when compared with the clinical data of patients"age,gender,tumor size,clinical stage,and histological grade;the incidence of methylation in 58 RCC tissues,paraneoplastic tissues,and normal tissues was 65.52%(38/58),34.48%(20/58),respectively.The positive rate of WNK2 gene methylation in RCC tissues was significantly higher than that in paraneoplastic and normal tissues(P<0.05),while the difference was not statistically significant when comparing paraneoplastic and normal tissues(P>0.05).There was no statistically significant difference between the incidence of methylation of WNK2 gene promoter region in RCC tissues and clinical data such as age,gender,tumor size,clinical stage,and histological grade of patients(P>0.05);the relative expression of WNK2 mRNA in the methylation group was significantly lower than that in the non-methylation group(P<0.05).Conclusions In RCC tissues the WNK2 methylation and the decline of WNK2 mRNA expression may be related to the development of RCC while they may not be related to the clinical progression of RCC.The decreased expression of WNK2 mRNA may be related to its promoter methylation.