Study On The Correlation Between Long Non-coding RNA And Mechanism Of Aldosterone-producing Adenoma

Part I Utilize CNC coexpression analysis to find related lncRNAsObjective:To find lncRNAs related to mRNAs:CYP11B2, PCP4, VPREB3 and PRRX1 which have greater difference between experiment group and control group in microarray.Methods:This study gained lncRNAs by means of CNC （coding-non-coding） coexpression with the criteria of PCC （Pearson correlation coefficient）>=0.85. Then these lncRNAs were checked in the differentially expressed lncRNAs in microarray to select target lncRNAs.Results:After the CNC coexpression analysis, this study gained 35,83,0 and 10 lncRNAs related to mRNA CYP11B2, PCP4, VPREB3 and PRRX1 respectively.7 target lncRNAs were selected through checking in the differentially expressed lncRNAs in microarray. They were NR₀03023, ENST00000417715, uc003zpr.2, ENST00000417262, CR605514, ENST00000503380 and ENST00000512207.Conclusion:There were 7 target lncRNAs:NR₀03023, ENST00000417715, uc003zpr.2, ENST00000417262, CR605514, ENST00000503380 and ENST00000512207.Part â…¡ Utilize real-time PCR to verify target incRNAsObjective:Real-time PCR was conducted on these 7 target IncRNAs in expanded samples [experiment group （aldosterone-producing adenoma+adrenal nodular hyperplasia）:33, control group （normal adrenal tissue）:19] to verify if there were still significant differences between experiment group and control group. It was hoped to find some IncRNAs able to explain the mechanism of APA.Methods:Total RNA was extracted from 33 experiment samples and 19 control samples. RNA was checked for its quality and integrity. Then the RNA was reversed transcribed into double strand cDNA. After adjusted by GAPDH, the 7 target IncRNAs were verified by real-time PCR with designed primers.Results:6 out of the 7 target IncRNAs were verified that the differences between experiment group and control group were significant and consistent with the results of CNC coexpression analysis and the microarray. They were:NR₀03023, uc003zpr.2, ENST00000417262, CR605514, ENST00000503380 and ENST00000512207. The real-time PCR result of lncRNA ENST00000512207 was consistent with the results of CNC coexpression analysis and the microarray but without significant difference. According to different pathologic types, this study divided experiment group into aldosterone-producing adenoma and adrenal nodular hyperplasia. Repeat the statistic analysis and our study got the same results. LncRNA uc003zpr.2 was the most specific lncRNA when our study graphed distribution map with experiment group and control group.Conclusion:LncRNA NR₀03023, uc003zpr.2, ENST00000417262, CR605514, ENST00000503380 and ENST00000512207 were verified that the differences between experiment group and control group were significant and consistent with the results of CNC coexpression analysis and the microarray. Among them, lncRNA uc003zpr.2 was the most specific lncRNA when our study graphed distribution map with experiment column and control column. It maybe strongly related with the mechanism of APA and was worth further investigation.