The Function And Mechanism Of Long Non-coding RNA RP13-650J16.1 In Prostate Carcinoma:an Preliminary Study

Objective:High-throughput microarray was used to screening the differentially between carcinoma tissue and normal tissue in prostate cancer,and select the key Lnc RNA.Nest,the key Lnc RNA was verified between carcinoma tissue and normal tissue in prostate cancer.Last,discussion the mechanism of the key Lnc RNA in prostate cancer.Methods:By differences in multiple,adjacent to genes encoding the information analysis,lenth of character analysis and other technique screening the key molecules which related to invasion and metastasis of prostate cancer.then we collected 15 clinical tissues from prostatectomy in our hospital and established the extraction methods of high-quality total RNA from clinical tissues by improve obtaining,transportation and grinding of samples.And detecting the expression of RP13-650J16.1 by q RT-PCR.Nest,we selecti the prostate cancer cell line 22RV1 and DU145 to culture.Construction of si RNA interference plasmid vector:si-584705.Three cells are divided into transfected group;negative control group,then used Lipofectamine TM2000 as si RNAs carrier transfections cells.Nest,we use q RT-PCR to authenticate wether our cells transfected.If 22RV1 or DU145 cell lines which inhibited expression of RP13-650J16.1 after transfection will detect the power of its proliferation,migration,and invasion by MTT,transwell and Tablet cloning experiments.And making the transfection of si-NC and control the negative control for comparison.Using bioinformatics to predict the target genes of RP13-650J16.1.Then we use q RTPCR to detect the effect of target genes after RP13-650J16.1 knock down.Finally the cells which transfected use Western blot detection the expression of downstream target genes regulating protein called RAC3 and BCL-2.The cells which transfected of si-NC as negative control group for comparison.Results:Microarray analysis showed that 13 lnc RNAs which associated with the cancer pathway have diversity of expression in prostate cancer,including lnc RNA RP13-650J16.1.Large organizations which used RT-PCR to detect its expression indicatethat the expression of lnc RNA RP13-650J16.1 in prostate cancer was significantly higher than adjacent tissue(p<0.05).Lower expression of RP13-650J16.1 which transfection by si RNA can lead to the lower of migration,invasion and proliferation capacity in prostate cancer.Through bioinformatic analysis of RP13-650J16.1 we find a gene which related prostate cancer called RAC3.It is a cancer gene in breast cancer.The result of RT-PCR and western blot showed that knock down the expression of RP13-650J16.1 will reduce the expression of m RNA and protein in RAC3.Conclusion:Through the qualcomm chips analysis,we found a group of lnc RNA molecules which have variance expression in carcinoma tissue and non-cancer tissue of prostate.RP13-650J16.1 is one of the key lnc RNAs which affected the invasion and metastasis in prostate cancer.And expression higher in prostate cancer tissue than non-cancer tissue.Inhibition the expression of RP13-650J16.1 can lead to less migration,invasion and proliferation in prostate cancer cells.RAC3 is the functional target gene of RP13-650J16.1.RAC3 promote the capacity of invasion and metastasis in prostate cancer.Inhibiting the expression of RP13-650J16.1 may lead to the lower expression of protein RAC3.So we think that RP13-650J16.1 may be a upstream molecules in RAC3 pathway.