The Expression Of Serum MiRNA In Colorectal Adenoma-carcinoma Sequence And Its Clinical Significance

Introduction Colorectal cancer is one of the most prevalent and deadly cancers, with an annual incidence of1million new cases.Colorectal adenomas were deemed to the main precancerous lesions for colorectal cancer, especially advanced adenomas. About60%-90%of colorectal cancer were evolved by colorectal adenomas. The time from adenoma to carcinoma requires about10-15years. Recent literature repeated that serum miRNA can be acted as potential molecular markers to diagnosis. The expression of many miRNAs is identified differentially in the process of adenoma to carcinoma. Therefore, searching miRNA in different stages of the evolution of colorectal adenomas towards carcinomas as noninvasive molecular markers is advantageous to the primary prevention for colorectal cancer and to understand the molecular mechanisms of colorectal adenomas towards carcinomas.Part ⅠThe expression of serum miRNA in patients with colorectal adenomas-carcinoma sequence analyzed by MicroarrayObjective:To identify serum miRNA differentially expressed in colorectal adenomas and early carcinomas patients. Methods:miRNA microarray was used to detect the differential expression of miRNAs in colorectal adenomas, early carcinomas and normal group.Resurts:In the condition of fold change>2.0or<0.5and P<0.05, there are21serum-specific miRNAs identified in colorectal adenomas, early carcinomas and normal group;15miRNAs identified between colorectal adenomas and early carcinomas;16miRNAs identified between colorectal adenomas and normal group;13miRNAs identified between early carcinomas and normal group;12miRNAs identified between all Adenomas (Ⅰ-Ⅱ) and all Adenomas Ⅲ. Among them, compared with normal group, serum miR-661is accurate higher expression in patients with colorectal adenomas; serum miR-591is accurate higher expression in patients with early cancer. Furthermore, the expression of miR-215and miR-193b-3p-miR-302b-3p was statistically significant in all adenomas (Ⅰ-Ⅱ)、Ⅲ and early carcinomas P<0.05).Conclusion:Serum miR-661and miR-591are respectively expected to become invasive molecular markers of colorectal adenomas and early carcinoma; miR-215, miR-193b-3p and miR-302b-3p may be play important role in the early stages of colorectal adenomas towards carcinomas.Part ⅡThe expression of serum miR-193b-3p, miR-150-5p and miR-204-5p validated by fluorescence quantitative PCRObjective:Validating the expression of serum miR-150-5p, miR-193b-3p and miR-204-5p in the patients in the phase of colorectal adenomas-carcinoma sequence by Real-time fluorescent quantitative PCR and preliminary discussing its relationship with the clinical pathological features.Methods:TaqMan (?) Human Microarray analysis was used to detect the expression of serum miRNAs in15colorectal adenomas patients,3colorectal cancer patients and3healthy controls. The serum miR-150-5p, miR-193b-3p and miR-204-5p, which were screened out form Microarray analysis, were validated in42patients in the phase of colorectal adenomas--carcinoma and5healthy controls by Real-time fluorescent quantitative PCR, as well as tentatively analyzed its relationship with the clinical pathological features. Results:In the condition of fold change>2.0or<0.5and P<0.05, the serum miR-150-5p, miR-193b-3p and miR-204-5p were screened out form Microarray analysis. miR-204-5p was differentially expressed in adenoma and early cancer and normal controls (P=0.02); miR-193b-3p was differentially expressed in low-grade intraepithelial neoplasia group (adenomas Ⅰ～Ⅱ level) and high-grade intraepithelial neoplasia group (adenomas III level+early carcinoma)(P=0.01); miR-150-5p was differentially expressed between villous adenoma and normal controls (P=0.02). Serum miR-204-5p validated by real-time fluorescence quantitative PCR were partially consistent with those detected by microarray. Serum miR-193b-3p, miR-150-5p expression level consistent with the Microarray analysis results, but there was no statistically significant difference. Also, compared with normal control group, serum miR-204-5p was lower expression in early cancer and advanced adenocarcinoma (P values respectively were0.02and0.01).Compared with adenoma group, miR-204-5p was lower expression in early cancer and advanced adenocarcinoma (P values respectively were0.01and0.002).Compared with normal control group, serum miR-150-5p was lower expression in advanced adenocarcinoma with significant difference (P=0.01).Compared with early carcinoma group, miR-204-5p was also lower expression in advanced adenocarcinoma with significant difference (P=0.03).Compared with normal control group, serum miR-193b-3p was lower expression in adenomas group with significant difference (P=0.02).Conclusion:Serum miR-193b-3p, miR-204-5p and miR-150-5p are respectively expected to become invasive molecular markers of colorectal adenomas,early carcinoma and advanced adenocarcinoma.miR-193b-3p and miR-204-5p may play important role in the early stages of colorectal adenomas towards carcinomas; miR-150-5p may play a role in the development of colorectal cancer.