| Background Esophageal squamous cell carcinoma(ESCC)was a common disease in human beings.It is mainly caused by the imbalance of multiple gene expression under the influence of internal and external factors and has a serious impact on the quality of life of the patients.Although the survival rate has improved in recent years,but the5-year survival rate of advanced esophageal cancer is s no more than 20%.Early lymph node metastasis,poor prognosis is an important character,the survival rates of esophageal cancer are still low,but its causes and mechanisms are still unclear.Therefore,looking for a early carcinoma marker of diagnosis and prognostic judgment for esophageal squamous is imminent.Previous study have been confirmed that miRNA is involved not only involved in the occurrence of esophageal squamous carcinoma,but also related to the invasion and metastasis of esophageal squamous carcinoma.With the deepening of the understanding of micro RNAs,will further clarify the role miRNA plays in esophageal squamous carcinoma,to provide a new basis for micro RNAs in the pathogenesis,early diagnosis,targeted therapy and judging prognosis in esophageal squamous carcinoma.However,the practical application shows that the miRNA test of tumor tissue is very traumatic,and the early diagnosis and therapeutic effect of the tumor can not be timely monitored.Compared with tissue miRNAs,serum miRNAs have the advantages of non-invasive,repeatable,and simultaneous detection of a number of indicators at the same time,its impact by external interference factors is small,strong clinical utility and potential as a valuable tumor marker.Objective To observe the expression of serum miR-30d-5p in patients of preoperative and postoperative in esophageal squamous cell carcinoma and the healthy control,to explore the relationship between serum miR-30d-5p and clinical pathological features as well as prognosis of patients,and to compare the expression differences of serum miR-30d-5p with serum TK1.In order to find a serum marker for early diagnosis and prognosis of patients with esophageal squamous carcinoma with high sensitivity and specificity.Methods Serum samples were collected from 30 patients with this kind of carcinoma from April to May in 2013.Microarray microarray was used to analyze the serum miRNA expression difference between(3 cases)of preoperative and postoperative sample.Screening of miRNAs was performed by Quantitative Real-time PCR,(q RT-PCR)validation to select new potential tumor-associated miRNAs for ESCC.The changes of onco-miRNA were examined before and after operation.Compare with the control group.Twenty-seven patients underwent preoperative and postoperative thymidine kinase 1(TK1)detection.Analyze the differences in expression before and after the operation of the target miRNA and TK1 in various clinical pathological parameters,and the postoperative expression level and prognosis of miRNA-30d-5p and TK1 were also analyzed by Cox regression,to find the independent risk factors related to the prognosis of ESCC.Results Microarray microarray was used to analyze the difference of miRNA expression between preoperative and postoperative group in small sample(3 cases).A total of 2555 miRNAs were detected,of which 143 were ≥500.Significant differences were observed in screening for miRNAs with large changes.The results of RNA oligonucleotide microarray showed that 21 miRNAs were significantly differentially expressed in the ESCC before and after resection of the cancerous tissue.There were 18 down-regulated miRNAs and 3 up-regulated miRNAs.Then the chip data into Cluster3.0 for cluster analysis.Mi R-30d-5p and miR-483-5p,which were significantly down-regulated after the surgery,were further validated by RT-PCR in combination with domestic and foreign literatures.The results of q RT-PCR showed that miR-483-5p had no significant difference before and after operation(P>0.05),but miR-30d-5p had the same trend as that of chip before and after operation(P<0.05).Therefore,miR-30d-5p was selected as a novel potential miRNA for ESCC.With q RT-PCR validation,serum miR-30d-5p level was significantly different before and after operation(P<0.05).The expression levels of miR-30d-5p were significantly higher in patients with esophageal squamous cell carcinoma than in the control group(P<0.05).The expression of serum miR-30d-5p levels in preoperative and postoperative patients with esophageal squamous cell carcinoma were significantly associated with the tumor depth of invasion,lymph node metastasis,tumor location and length,histopathological type and degree of differentiation,as well as history of cigarette smoking and drinking(P<0.05).Meanwhile,TK1 in preoperative and postoperative patients with esophageal squamous cell carcinoma was associated with depth of tumor invasion,histopathological type and degree of differentiation,as well as history of cigarette smoking and drinking(P<0.05).The results showed that the change of serum miRNA levels(mean number)in serum was better than TK1 in ESCC.Patients with lower miR-30d-5p expression had longer disease free interval(DFI)and overall survival(OS)than patients with higher miR-30d-5p expression.Univariate analyses confirmed that lower expression of miR-30d-5p both in esophageal squamous cell carcinoma tissue is associated with longer DFI and OS.Conclusion Mi R-30d-5p has the potential for tumor markers of esophageal squamous carcinoma.High expression of miR-30d-5p having a certain promoting role in the occurrence and development of esophageal cancer and predict poor prognosis.Serum miR-30d-5p can be used for early diagnosis and prognosis judgment,maybe more sensitive than serum TK1. |
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