Differentially Expression Of Long Non-coding RNAs In The Malignant Transformation From Proliferative Inflammatory Atrophy To Prostate Carcinoma: An Preliminary Study

Objectives:To analyze the differentially expressed m RNA and lnc RNA among prostatic carcinoma(PCa),Proliferative inflammatory atrophy(PIA)and their paired adjacent noncancerous tissues by microarray, with the aim to detect genetic alterations in neoplastic transformation and identify the possible distinctive network of prostatic nonresolving inflammation transformation into prostate carcinogenesis for refreshing molecular mechanism and providing new proposal in the preservation, clinical diagnosis, treatment and pathogenesis of prostate cancer. Methods:First of all, we collected 20 clinical tissues from prostatectomy between August 2013 and August 2014 in our hospital and established the extraction methods of high-quality total RNA from clinical tissues by improve obtaining, transportation and grinding of samples. Then, the purity of PCa, PIA and their paired adjacent noncancerous clusters in the same prostate were isolated by cryosections combined with manual microdissection technology. In the end, we performed Bioinformatic analysis of them by combining lnc RNA expression profiling and m RNA expression profiling. The Pathway analysis and targets prediction using reversing Cis were performed, meanwhile, we constructed a genomic network of lnc RNA and its target genes in some inflammation and/or cancer signaling pathways, related to the differentially expression profiling. Results:1. Tissues of radical prostatectomy were put and cryopreserved in liquid nitrogen within 30 minutes.then, high-quality total RNA(RIN>7.5)might be extracted from those samples through liquid nitrogen grinding or cryosections.2. Tissues obtained from manual microdissection were used for high-quality total RNA extraction by Trizol, which could meet the standards of high-throughput microarray technology, but H&E staining may destroy the integrity of the RNA.3. Of the 2610 m RNA dysregulated detected in the human PCa tissues when compared to adjacent noncancerous tissues, 689 genes were up-regulated and 1921 genes were down-regulated. At the same time, 2176 lnc RNA were differentially expressed, with 688 up-regulation and 1488 down-regulation. Compared with adjacent noncancerous tissues, 183 of 592 m RNA profiles were up-regulated and 409 of 592 were down-regulated in human PIA tissues, meanwhile, 575 lnc RNA were differentially expressed, with 160 up-regulation and 415 down-regulation. Of 1007 m RNA dysregulated between the PCa tissues and the PIA tissues, 115 genes were up-regulated and 892 genes were under-regulated.773 lnc RNA were differentially expressed as well,105 lnc RNA of them were up-regulated and 668 lnc RNA of them were under-regulated. The 113 overlapping m RNA showed significant linear difference among them, 8 genes were commonly up-regulated and 105 genes were commonly down-regulated. Also, The 106 overlapping lnc RNA showed significant linear difference among them, 11 genes were commonly up-regulated and 95 genes were commonly down-regulated.4. Pathway analysis of the differentially expressed genes revealed that 22 signaling pathways were the top over-represented terms, nine of them closely correlated with inflammation and/or cancer, which altered dramatically in the progress of the disease. Conclusions:1. It is an economical and valid method to qualify and isolate targeted sub-lines by H&E staining of cryosections combined with the “elimination” manual microdissection, which were used for extracting high-quality RNA consistent with the standards of high-throughput microarray technology.2. Gene expression profile chip is an ideal technology for screening m RNA and lnc RNA dysregulated.3. Genes significantly differentially expressed were detected among PCa, PIA and their paired adjacent noncancerous clusters. masses of differential molecules interacted with each other and involved in the signaling pathways in inflammation and/or cancer, which prognosticated that lnc RNA may play a critical role in neoplastic transformation resulted from nonresolving inflammation. Lnc RNA may become novel tumor biomarkers and potential targets of therapy.