MiR-181a Regulates Radiosensitivity In Cervical Cancer Through Targeting PRKCD

Purpose:to study the relation between miRNA expression and radiosensitivity in cervical cancer, we aim at finding certain miRNA to regulate radiosensitivity and showing regulatory network.Methods and Materials:We hypothesize that miRNAs interconnect with mRNA involved in radiosensitivity, which influence intrinsic radiosensitivity of tumor, we obtain the miRNAs associated likely with radiosensitivity though screening miRNAs expression profiling in radiosensitive and radioresistant human cervical cancer sample using microarray. In order to test the result of microarray, Real-time Polymerase Chain Reaction is performed. Cervical cancer cell, SiHa and Me180, are transfected by miR-181a-mimic, miR-181a-inhibitor.Stably transfection cell lines are constructed by Lentivirus vector carrying miR-181a. Colony-Formation Assay was performed to detect the radiosensitivity. Through cell proliferation assay, cell cycle assay and apoposis assay, we searched for the molecular mechanism which miR-181a depressed radiosensitiviy. To find the mRNA expression profile of stably expressing miR-181 cells SiHa and Me180 were analyzed by way of mRNA microarray. Furthermore, two prediction algorithms PicTar and TargetScan were used to analyze the possible target gene. Quantitative reverse transcription polymerase chain reaction, Real-time Polymerase Chain Reaction and Western analysis were performed to study the target gene of miR-181a. Finally, Restoring the expression of target gene, PRKCD, in cells expressing miR-181a could reverse the radiation protective effect of miR-181a.Results:MiRNA microarray shows that two miRNAs(miR-181a, miR-21) are overexpressed in the radioresistant samples compared to the radiosensitive samples, and six miRNAs (miR-30*, miR-23a, miR-16-2*, miR-378, miR-18a, miR-221) are underexpressed. The overexpression of miR-181a can inhibit the radiosensisitivity of cervical cancer in vitro and in vivo. The mRNA expression profile of stably expressing miR-181 cells SiHa and Me180 were analyzed by way of mRNA microarray. Furthermore, two prediction algorithms PicTar and TargetScan were used to analyze the possible target gene. Integrating three methods, PRKCD becomes the candidate. Knockdown of PRKCD protected against radiation damage in cervical cancer cells. Restoring the expression of PRKCD in cells expressing miR-181a could reverse the radiation protective effect of miR-181aConclusions We conclude that miR-181a regulates radiosensitivity in cervical cancer through targeting PRKCD.