Expression And Methylation Status Of LOC100130476 In Renal Cell Carcinoma

Objective:Renal Cell Carcinoma(Renal Cell Carcinoma,RCC)is one of the most common malignant tumors in the urinary system,which originates from renal parenchyma urinary tubular epithelium.It is now a serious threat to the health of our population.In recent years,because the early symptoms of renal cell carcinoma are not obvious and nonspecific,the mortality rate is rising.Besides the imaging examination,renal cell carcinoma is still lack of early laboratory diagnosis index,so early diagnosis and treatment of renal cell carcinoma are difficult.Nowadays,the research on the pathological mechanism of tumor has been paid more and more attention,which found that epigenetics is a very important part in the process of tumor formation and development.Research on epigenetics includes DNA methylation,histone modifications,genomic imprinting,maternal effects,gene silencing,chromatin remodeling,genetic imprinting and random chromosome(X)inactivation and non-coding RNA regulation and so on.Recently,the relationship between DNA methylation of tumor suppressor genes and tumor formation and progression is one of the focuses in the field of cancer research.It has been confirmed that methylation of the first exon of LOC100130476 gene is an important factor leading to the occurrence and progression of gastric cancer,esophageal cancer and other malignant tumors.This research studied 57 postoperative specimens of patients with RCC,in which the relationship between the methylation status of the first exon of LOC100130476 gene and the relative expression of its mRNA in the RCC tissues and normal kidney tissues was analyzed to investigate the correlation between LOC100130476 and renal cell carcinoma and its possible mechanism;It further revealed the pathogenesis of RCC,which provided a theoreticalbasis for early diagnosis,treatment and prognosis evaluation of renal cell carcinoma.Methods:In this study,methylation specific PCR(Methylation specific PCR)method was used to detect the methylation status of the first exon of non coding long chain RNA LOC100130476 gene in 57 cases of RCC tissues and normal tissues;Reverse Transcriptase Polymerase Chain Reaction(Reverse Transcriptase-PCR,RT-PCR)was used to detect the relative expression of mRNA of LOC100130476 gene in RCC tissues,and Statistical software IBM SPSS STATISTICS was used to analyze the relationship between LOC100130476 gene and clinical parameters.Results:1 Analysis of methylation status of LOC100130476 gene in RCC tissues and normal tissuesThe methylation status of LOC100130476 gene can be divided into three types: complete methylation state,incomplete methylation state and complete non methylation state,according to the results of MSP electrophoresis.Methylation positive contains complete methylation status and incomplete methylation state,while methylation negative is only completely non methylated state.The methylation rate of LOC100130476 gene in 57 cases of RCC was 66.7%(38/57),but the rate was only 26.3%(15/57)in the normal tissues.The results showed that the methylation rate of LOC100130476 gene in RCC tissues was significantly higher than that in normal tissues(P<0.05).2 Analysis of the relationship between the methylation status of LOC100130476 gene in the first exon of RCC gene and clinical dataThe data were grouped by age,sex,tumor size and analyzed statistically.There was no significant difference in the incidence of methylation of the first exon of LOC100130476 gene(P>0.05);however,there was significant difference in histological grade and TNM stage(P<0.05).The rate of methylation in the poorly differentiated group was higher than that in the high and middle differentiation group,and the methylation rate was higher in patients with TNM stage ?-?than that in patients with TNM stage?-?.3 Analysis of the relative expression of the mRNA of LOC100130476 gene in RCC tissues and normal kidney tissuesThe relative expression of LOC100130476 gene in RCC tissues was0.384±0.07,while the relative expression of the mRNA of LOC100130476 gene in the normal tissues was 0.596±0.06.The results showed that the relative expression of the mRNA of LOC100130476 gene in RCC tissues was significantly lower than that in normal tissues,and the difference was statistically significant(P<0.05).4 Analysis of the relationship between the relative expression of the mRNA of LOC100130476 gene in RCC tissues and clinical dataWhen the data were grouped by age,sex,tumor size and analyzed statistically,there was no significant difference in the relative expression of the mRNA of LOC100130476 gene(P>0.05),However,when the data were grouped by the histological grade and the clinical stage of tumor,there was a statistically significant difference in the relative expression(P<0.05).5 In RCC group,the relative expression of the mRNA of LOC100130476 gene in methylation group was significantly lower than that in non methylation group(P<0.05),the difference was statistically significant.Conclusions:1 In RCC tissues,the hypermethylation of the first exon of LOC100130476 was frequent,and the methylation rate was significantly higher than that of normal tissue,which suggested that aberrant methylation of LOC100130476 may be one of the mechanisms of renal cell carcinoma.2 In RCC tissues,the relative expression of the mRNA of LOC100130476 gene was significantly lower than that in normal tissues,which suggested that LOC100130476 might play an important role in the development of renal cell carcinoma as a tumor suppressor gene.3 In RCC tissues,the relative expression of the mRNA of LOC100130476 gene in patients with positive methylation was significantly lower than that in patients with negative methylation,which suggested that the high methylation status of the first exon of LOC100130476 gene might be animportant reason for the down-regulation of expression.