Study On Cell Carcinoma And Surgery Evaluation Of Esophageal Squamous MicroRNA As Serum

Esophageal cancer is a kind of malignant tumour of digestive tract, which the incidence of disease account for eighth, the death rate accounts for sixth in the worldwide. The incidence in men is higher than women. Esophageal cancer can be divided into esophageal squamous cell carcinoma (ESCC), esophageal adenocarcinoma (EAC) and undifferentiated carcinoma. China is one of the high-risk areas. The early symptoms of esophageal cancer are not obvious, it is mostly in the middle-late stage after diagnosis, and prognosis effect of ESCC patients is complex. It is difficult to improve the survival rate of patients effectively. The5-year survival rate is20%to30%for patients who have curative surgery and do not develop lymph node metastases. However, the survival rate is only13%for patients with lymph node metastasis. Consequently, it is emergent to look for diagnosis and prognostic markers of ESCC. MicroRNAs (miRNAs) are small, endogenous non-coding RNAs which are19-24nucleotides in length; they can posttranscriptionally regulate gene expression through partial base-pairing with the3’-untranslated region of target mRNAs, resulting in mRNA destabilization and translational inhibition. The studies show that the expression of some miRNA was changed obviously in tumor tissue. In addition, a distinctive expression pattern of serum miRNAs in serum from early stage cancer such as prostate cancer, colon cancer, pancreatic cancer and esophageal cancer has also been reported. So serum miRNA can be a new potential biomarker in patients’ early diagnosis and prognosis. Then we suspected that distinctive expression miRNAs in serum could be used as a biomarker of ESCC patients for diagnosis, surgical treatment and prognosis markers.Objectives:In this study, we screened the serum miRNA significantly increased in serum of ESCC patients and significantly decreased after surgery in order to evaluate its clinical application value of ESCC patients.Methods:Serum samples were taken from63ESCC patients with preoperative and postoperative and63age-and gender-matched controls.26of the ESCC patients contain different time point serum samples after operation for the study of prognosis. First, an initial screen of miRNA (inciude768miRNA) expression was performed in serum samples from28ESCC patients and28controls by the low density arrays. Screen the significantly up-regulated serum miRNAs in ESCC patients. Second, using real-time fluorescent quantitative PCR technique (quantitive real-time PCR, qRT-PCR) to detect some low density array screened miRNA, part of the increased significantly serum miRNA found in our early research and digestive system tumors related miRNA reported before in the screen group (18ESCC preoperative, postoperative serum samples and18normal serum samples) one by one, screen the significantly increased serum miRNA. Then use qRT-PCR technology in the validation group (45cases of ESCC patients preoperative, postoperative serum and45cases of normal serum) to screen miRNA associated with prognosis in the screen and validation set, in order to evaluate the surgical efficacy in patients with ESCC. The different time points after surgery in patients with26cases tracking detection, according to the visit of patients after surgery and statistical methods to evaluate its value in predicting survival condition.Results:The low density arrays results showed that serum miRNA expression profile in ESCC patients were significant different from controls and21serum miRNAs were markedly up regulated in ESCC patients compared to controls. We based on preliminary research, the digestive system tumors relate miRNA as well as low-density arrays results, screened out of11miRNAs (miRNA-7,25,31,100,141,1247,193a-3p,194,223,337-5p and483-5p) as the object of our study. qRT-PCR identified a profile of nine serum miRNAs (miR-25,31,100,193a-3p,223,194,141,337-5p and miR-483-53p) to be distinctive expressed (with the fold change ranged from1.57-6.42, P<0.05) for ESCC in training set include18ESCC and18controls. This results were further confirmed in another45ESCC and45controls with the fold change ranged from1.47-8.09(P<0.05). Five miRNAs (miR-141,193a-3p,194,223and miR-337-5p) were screened in total63ESCC samples which were significantly reduced in postoperation than preoperation. The area under the receiver operating characteristic (ROC) curve ranged from0.716to0.878for each selected ESCC specific miRNA:miR-25(AUC=0.845,95%CI=0.775-0.914), miR-31(AUC=0.748,95%CI=0.663-0.833), miR-100(AUC=0.785,95%CI=0.705-0.866), miR-141(AUC=0.716,95%CI=0.626-0.806), miR-193a-3p (AUC=0.861,95%CI=0.796-0.927), miR-194(AUC=0.827,95%CI=0.755-0.899), miR-223(AUC=0.778,95%CI=0.699-0.857), miR-337-5p (AUC=0.878,95%CI=0.812-0.943), miR-483-5p (AUC=0.84,95%CI=0.79-0.89), which was significantly higher than that of the carcinoembryonic antigen (AUC=0.578,95%CI=0.473-0.682). Using the optimal cut-off value, the sensitivity and specificity for ESCC-pre miRNAs were71%and86%for miR-25,62%and81%for miR-31,68%and81%for miR-100,50%and86%for miR-141,75%and89%for miR-193a-3p,62%and90%for miR-194, and65%and79%for miR-223,78%and91%for miR-337-5p, and75%and82%for miR-483-5p respectively. More importantly, miR-223in samples with different postoperation time has a significant positive correlation with the patients’ condition and living conditions.Conclusions:9miRNA (miR-25,31,100,141,193a-3p,194,223,337-5p and miR-483-5p) in preoperative serum samples of ESCC patients have significant difference with normal controls and has the potential diagnostic value.5miRNA (miR-141,223,337-5p,194and miR-193a-3p) in postoperative serum samples of ESCC patients reduced significantly, has the potential evaluation of operation effect. And miR-194,141expressed in died patients two weeks after operation is reduced, and rised in the serum in different time points after surgery, has the potential to assess ESCC patients condition. MiR-223in different time points after surgery changed significantly and associated with disease progression in patients, has the ability to assess prognosis.