A Preliminary Study On The Function And Mechanism Of Circ＿0007142 In Colorectal Cancer

Objective: To investigate the biological function and mechanism of circular RNA circ＿0007142 in colorectal cancer cells,and to clarify its important role in the development of tumor,which would provide a better target selection for the molecular targeted therapy of tumor.Methods:(1)A series of differentially expressed circRNAs in colorectal cancer were obtained from 6 colorectal carcinoma and corresponding paracancerous tissues.Based on the bioinformatics prediction software,five possiblely targeted microRNAs(miRNAs)of each circRNA and their binding sites were obtained.(2)Quantitative real-time PCR(qRT-PCR)assay was performed to analyze the expression of circ＿0007142 in fifty paired CRC tissues and paracancerous tissues.Correlation between circ＿0007142 expression and patients’ clinicopathological features was identified.(3)qRT-PCR assay was used to detect the expression of circ＿0007142between CRC cells(HT-29、HCT-116、LOVO)and normal colorectal epithelia cell(HCO).(4)The expression level of circ＿0007142 was downregulated with siRNA(Small interfering RNA,siRNA)in HT-29 and HCT116 cells,and the effects of circ＿0007142 on CRC cells’ proliferation,migration and invasion were studied.(5)QRT-PCR assay and dual luciferase reporter assay were performed to validate five highest scoring miRNAs predicted by Arraystar’s home-made miRNA target prediction software based on Target Scan and miRanda.Results:(1)The expression of circ＿0007142 in CRC was up-regulated,which was consistent with the results of the circRNA microarray.The analysis of the correlation between circ＿0007142 and clinicopathological features of the patients showed that the expression of circ＿0007142 was related to the lymph node metastasis(P =0.037)and the degree of differentiation of the tumor(P =0.008)in CRC patients.(2)In different cell lines of CRC,the expression level of circ＿0007142 was significantly higher in HT-29 and HCT116 cells than in HCO.(3)Downregulation of circ＿0007142expression in HT-29 and HCT-116 cells can result in a significant decrease in the ability of proliferation,migration and invasion of CRC cells;(4)The expression of miR-103a-2-5p in thirty CRC tissues and paired adjacent normal tissues was down regulated,which was negative correlated with circ＿0007142 in shared thirty tissues.Up-regulation of circ＿0007142 was observed in CRC cells with siRNA disposed.Further dual luciferase reporter assay also come to the conclusion that circ＿0007142and miR-103a-2-5p could interact with each other.Conclusion : Circ＿0007142 was significantly upregulated in CRC,which could promote the proliferation,migration and invasion of CRC cells.Circ＿0007142possiblely play a genetic regulatory role through miR-103a-2-5p.