MicroRNA-124Regulates The Proliferation Of Colorectal Cancer Cells By Targeting IASPP

Objective:The goal of the study is to test whether miR-124inhibits proliferation of colon cancer cells by targeting iASPP.Methods:1, All specimens were taken from patients undergoing colon cancer operation. Subjects include18males and22females. None of the patients had taken radiation or chemotherapy before surgery. The age ranged from35to76years, and the average age was50.27years. By UICC and AJCC2012TNM staging of colon cancer and the clinical stage,11cases are diagnosed as stage Ⅰ,9cases as phase Ⅱ,20cases as stage Ⅲ. Normal colon tissue of the same patient was takenfor control.2, Normal human colonic epithelial cells were selected as control. Three colorectal cancer cells (SW480,HT29,SW620) were selected as the experimental group. iASPP was detected by QPCR to predict miRNAs expression.3, SW480and HT29cells were transfected by miR-124mimics and iASPP-shRNA. Expression of iASPP and NF-к b were tested by Western blot. iASPP was overexpressed to reply miR-124’s affect on iASPP and NF-Kb expression;4, Dual-luciferase report gene was applied to further verify the miR-124target regulation of iASPP;5, miR-124Lentiviral vectors was built to transfect HT29cells and SW480. MTT, Brdu and cloning method was used to detect effect of miR-124on HT29and SW480cells proliferation. Overexpression of iASPP was used to reply biological effects of miR-124on HT29and SW480cells.Result:1, On the level of organ, QPCR showed in various forecasting target-regulating miRNAs of the iASPP, miR-124was specificly low expressed in colon cancer tissue. On level of cell, QPCR also showed that miR-124was specificly lowly expressed in colon cancer cells;2, When miR-124, analogue and iASPP-shRNA were transfected to SW480and HT29cells to overexpress miR-124or silent iASPP, expression of IASPP was significantly inhibited, while expression of NF-Kb was upregulated; After iASPP ORF clone was transfected, overexpression of iASPP could reply miR-124’s regulation on iASPP and NF-Kb;3, After miR-124Lentiviral vectors were built and transfected to HT29and SW480cells, GFP fluorescence picture showed infection efficiency of more than80%. Cell function results showed overexpression of miR-124or silented iASPP can significantly inhibit cell proliferation.Conclusion:1, Among miRNAs targeting iASPP predicted by authoritative software, miR-124was specificly lowly expressed in human colon cancer tissue and cell cancer cells;2, miR-124can inhibit the expression of iASPP and upregulate the expression of NF-Kb;3, Overexpression of miR-124or scilencing iASPP can significantly inhibit colon cancer cell proliferation;4, These results showed miR-124inhibit colon cancer cell proliferation through targeting iASPP.