| Along with the social structure of the Chinese population aging,the incidence of malignant tumors around the country is on rise year by year?According to the 2012 China cancer registration update issued by the national cancer center,the number of new tumor disease cases is about 3.12 million every year,which suggests that an average of six people are diagnosed with malignant tumor every minute.Morbidity and mortality squamous of esophageal cancer in common cancers are respectively the eighth and the sixth,of which cell carcinomas(esophageal squamous cell carcinoma,ESCC)occupied the main position in East Asia,and about 50%of esophageal squamous carcinoma cases occur in China.China is a country with a high incidence of esophageal cancer,and more than 90%of the pathological types of esophageal cancer are squamous cell carcinoma.However,the early symptoms of esophageal squamous cell carcinoma are not obvious,and most patients are often diagnosed at the middle or advanced stage.At present,the treatment of esophageal squamous cell carcinoma is relatively limited.The treatment effect is not ideal,and the overall 5-year survival rate for patients is only 15%to 25%.Therefore,the early diagnosis of esophageal squamous cell carcinoma is of great significance for optimizing the treatment and diagnosis of esophageal squamous cell carcinoma.MicroRNAs(miRNAs)are non-coding small nas that can regulate gene expression at post-transcriptional level.MiRNA can degrade or inhibit the transcription of mRNAs by binding to the coding region or 3'-UTR of target mRNAs.In recent years,with the development of molecular biology,more and more miRNAs have been found to play an important role in the development of tumors.In this study,on the basis of the present researches,we selected two kinds of miRNAs(miR-373-3p and miR-193-b-3p)as the research object,miR-373-3p was up-regulated in a variety of tumors,and was considered to cancer gene play a role in the tumor;miR-193b-3p is reduced in tumor and is often seen as a tumor suppressor gene.In the esophageal squamous cell carcinoma,we studied the two miRNAs,hoping to explain the mechanism of miRNA in esophageal squamous cell carcinoma more comprehensively.We detected the 2 kinds of miRNAs' the expression levels and relevant genes' expression levels in serum and tissue of the patients with esophageal squamous cell carcinomas by RT-qPCR,and with the MSP method we detected the degree of methylation gene,to initially explore two miRNAs' expression in esophageal squamous carcinoma and miRNAs' molecular regulation mechanism.The results are as follows:1.The expression levels of miR-373-3p and miR-193b-3p were detected by rt-qpcr,and the high expression of mir-333-3p in esophageal squamous cell carcinoma was detected(p<0.01),and mir-193b-3p was low in esophageal squamous cell carcinoma(p<0.01).And the AUC values of 2 kinds of miRNAs were 0.883(95%CI:0.776-0.989;Sensitivity is 80%;Specificity is 90%);0.815(95%CI:0.679-0.937;Sensitivity is 81.7%;Specificity is 73.7%).Moreover,we found that the expression levels of two serum miRNAs were inconsistent serum miR-373-3p preoperative high expression,postoperative low expression;serum miR-193b-3p preoperative low expression and postoperative high expression were significantly different(p<0.01).Comparision of two kinds of miRNAs levels in cancer tissues and adjacent normal tissues of ESCC,the expression level of miR-373-3p in cancer tissues was significantly higher than adjacent tissues,and the expression level of miR-193b-3p in cancer tissues was significantly lower than adjacent tissues,which was the same as the previous experimental trend in serum.Therefore,miRNAs in serum may be from ESCC cancer tissues.These results suggest that the two kinds of miRNAs selected may be used for early diagnosis and treatment of esophageal squamous cell carcinoma.2.Regulation mechanism of mir-373-3p.According to the prediction results of bioinformatics and the relevant literatures,we selected the Hippo signaling pathway and p53 signaling pathway related to miR-373-3p.And the NF2.LATS2,TAZ,JNK,OXR1,CASP9 gene mRNA expression level is analyzed,finding 6 gene expression level in esophageal squamous carcinoma and miR-373-3 the expression level of miR-373-3p is a negative correlation,miR-373-3p and target genes OXR1 and LATS2 correlation coefficient r=-0.699,-0.704,respectively,with statistical significance(p<0.05).Preliminary results showed that miR-373-3p participates in the development of esophageal squamous cell carcinomas by influencing the Hippo and p53 signaling pathway related target genes and its related gene expression·3.Regulation mechanism of miR-193b-3p.In the same way,we also selected two target genes of miR-193b-3p and their upstream and downstream genes,and analyzed the expression levels of 6 genes of RB,CCND1,P16,IGF1,IGF1R and PI3K.The results showed that CCND1,IGF1,IGF1R and PI3K were highly expressed in cancer tissues,and RB and P16 were low expression in cancer tissues.The correlation coefficient of miR-193b-3p and genetic CCNDI is r=-0.645,with statistical significance(p<0.05),indicating that miR-193b-3p may affect the expression levels of target gene and its upstream and downstream genes to participate in the occurrence of esophageal squamous carcinoma.4.In order to explore the relationship between the methylation degree of miR-373-3p and miR-193b-3p gene promoter region and its expression level,the study tested the degree of methylation of miR-373-3p and miR-193b-3p gene promoter region in the patients with esophageal squamous carcinoma,and found that miR-193b-3p gene promoter region tend to be high methylation and miR-373-3p gene promoter region tend to be low methylation;the lower the miR-373-3p gene promoter methylation is in patients,the higher the expression level of the miR-373-3p is,and the higher the miR-193b-3p gene promoter methylation is in patients,the lower the expression level of miR-193b-3p is.The preliminary experimental results show that there could be a negative correlation between the degree of methylation in the miRNAs genes promoter region and the expression level of miRNAs.Combined with the experimental results,this study analyzed the possible mechanism of miR-373-3p and miR-193b-3p in the esophageal squamous carcinoma.Namely the methylation of gene promoter region can lead to elevated expression of miR-373-3p and low expression of miR-193b-3p,so that carcinogenic factor enhanced,tumor suppressor factor decreased which affects the whole channel system and lead to the occurrence of tumor development. |
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