| Esophageal cancer(EC) is one of the most common human malignancies. Esophageal squamous cell carcinoma(ESCC) and esophageal adenocarcinoma(EAC) are the two main histological types. ESCC is the main subtype of EC, mostly occurring in Asia and Africa, whereas EAC mostly occurs in western countries. The prognosis of EC patients is poor, with an overall 5-year survival rate of 15%-25%, which, however, can be promoted to more than 90% for early EC patients. Micro RNAs(mi RNAs) is a kind of single stranded non-coding RNAs with only 17-25 ribonucleotides, which can inhibit target m RNAs translation or induce their degradation by binding to the 3'-untranslated region(3'-UTR) of target m RNAs through base-pairing manner. mi RNAs are estimated to regulate the expression of almost 30%-60% of human genes. They are demonstrated to be involved in various cellular biological and pathological processes including proliferation, differentiation, apoptosis, cell cycle, invasion and migration, thus playing important roles in the development and progression of a lot of cancers. Circulating mi RNAs are the mi RNAs that exist in the peripheral blood, including plasma or serum mi RNAs according to the sample type. Accumulating evidence demonstrated that circulating mi RNAs are dysregulated in numerous cancers, and present great clinical application value in the diagnosis, therapy and prognosis. Circulating mi RNAs are thought to originate from the tumor tissues. Their expression level is in accordance with that of the tumor tissues. What's more, they are not only resistant to RNase digestion but also to acid/alkali catalysis, and were less subject to retention time and temperature changes, thus can stably exist in the peripheral blood. Therefore, circulating mi RNAs are thought to be a kind of perfect tumor biomarkers. Mi RNA-216a/b are two members of mi RNA-216 family. Recent years, they were demonstrated to be dysregulated and involved in the development and progression of many human cancers, such as non-small cell lung cancer(NSCLC), colorectal cancer(CRC), hepatocellular carcinoma(HCC), oral spuamous cell carcinoma(OSCC), and nasopharyngeal carcinoma(NPC). Moreover, plasma or serum mi RNA-216a/b presents potential value in the diagnosis of acute pancreatitis and pancreatic cancer(PCA). However, the role of mi RNA-216a/b in ESCC has never been reported previously. The present study was designed to examine the difference of plasma mi RNA-216a/b expression level between ESCC patients and the healthy controls, and between the pre- and post-operative ESCC patients. Then the correlation of plasma mi RNA-216a/b with the clinicopathological characteristics of ESCC patients was analyzed and the diagnostic value of plasma mi RNA-216a/b for ESCC was evaluated, hoping to provide some valuable information in the early diagnosis of ESCC.Methods 1 Subjects: Experimental group: A total of 120 consecutive patients of ESCC at the First Affiliated Hospital of Zhengzhou University were recruited and all patients' clinicopathological characteristics were collected. Control group: 51 individuals, who sought a routine health check-up and did not have any esophageal diseases or other cancerous diseases, were recruited at the same period. 2 Plasma samples collection and storage: The peripheral blood samples of 120 ESCC patients(at the second day after admission), 21 ESCC patients who underwent esophagectomy and 51 healthy controls were collected. Once collected, the peripheral blood samples were centrifuged to isolate the plasma samples. Then all plasma samples were stored at-80 ? until further process. 3 Detection of the mi RNAs expression levels: Quantitative real-time PCR(q RT-PCR) method was performed to detect the expression levels of plasma mi RNA-216a/b and the endogenous reference gene mi RNA-16. The 2-?Ct(?Ct=Cttarget-Ctmi RNA-16) method was performed to calculate the relative expression levels of mi RNA-216a/b. 4 Statistical analysis: SPSS statistics software version 17.0(SPSS Inc., USA) was used for the statistical analysis. All continuous data were presented as mean±SD and categorical variables were presented as counts and percentage. The unpaired Student's t test or Mann–Whitney U-test was performed to compare the difference of mi RNAs expression levels between two groups. The paired t test was utilized to compare the difference of mi RNAs expression levels between paired groups. The one-way ANOVA or Kruskal–Wallis test was performed to compare the difference of mi RNAs expression levels among three or more groups. Receiver operating characteristic(ROC) curve and the area under the ROC curve(AUC) were used to evaluate the diagnostic power of plasma mi RNAs for ESCC. All tests were two-sided, and the significance level was set at P<0.05.Results: 1 The expression level of mi RNA-216a/b was significantly downregulated in ESCC patients compared with that of healthy controls(0.068±0.052 vs 0.179±0.098, P<0.0001;0.091±0.087 vs 0.199±0.161, P<0.0001) 2 The expression level of plasma mi RNA-216 b in postoperative samples was significantly upregulated compared with that of preoperative samples(0.132±0.037 vs 0.088±0.051, P=0.0074). Although the expression level of plasma mi RNA-216 a was also upregulated after esophagectomy, the result was insignificant(P=0.2619). 3 Patients with lymph node metastasis showed significantly lower mi RNA-216 b expression level than patients without lymph node metastasis(0.068±0.054 vs 0.134±0.117, P=0.0168), while the mi RNA-216 a expression level showed nosignificant difference(P=0.0845); patients with TNM ? showed significantly lower mi RNA-216 a expression level than patients with TNM 0-? or TNM ?(0.039±0.022 vs 0.088±0.062, P<0.05; 0.039±0.022 vs 0.086±0.065, P<0.05); patients with TNM ? showed significantly lower mi RNA-216 b expression level than patients with TNM 0-?(0.072±0.067 vs 0.134±0.119, P<0.05). There was no significant difference of mi RNA-216a/b among other clinicopathological characteristics including age, sex, smoking, alcohol, tumor location and histologic grade. 4 ROC curve analysis was performed to evaluate the diagnostic value of plasma mi RNA-216a/b for ESCC. At the cut-off value of 0.070, the AUC was 0.877(95% CI:0.818-0.922), the sensitivity was 80.0% and the specificity was 90.2% for mi RNA-216 a. At the cut-off value of 0.060, the AUC was 0.756(95% CI: 0.685-0.819), the sensitivity was 55.8% and the specificity was 90.2% for mi RNA-216 b.Conclusion: The expression levels of plasma mi RNA-216a/b were downregulated and presented satisfactory clinical application value in the detection and surgical treatment evaluation of ESCC. Additionally, the expression levels of plasma mi RNA-216a/b were inversely correlated with lymph node metastasis and/or TNM stage of ESCC patients, indicating that mi RNA-216a/b might play suppressive roles in the development and progression of ESCC. |
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