Preliminary Study On Pseudogene And Lncrna As Non-Invasive Biomarkers In Clear-Cell Renal Cell Carcinoma

Objective:To preliminarily investigate the possibility of pseudogene and long non-coding RNA(LncRNA)as non-invasive biomarkers for clear cell renal cell carcinoma(ccRCC).Methods:Combined with the previous research of our research group and reviewed the relevant literature,we selected eight LncRNAs and ten pseudogenes,the expression difference of which was preliminarily tested using the plasma from patients with ccRCC and healthy adults.The specific steps included extracting total RNA from plasma,reverse transcription to cDNA,using 5sRNA as internal reference and real-time fluorescence quantitative PCR.After preliminary test,1 LncRNAs and 4 pseudogenes with significant expression difference in plasma between patients with ccRCC and healthy adults were screened out.Subsequently,we used similar methods and a large sample size including plasma samples from 55 patients with ccRCC and 26 healthy adults to validate and screen,in order to determine the potency of these LncRNAs and pseudogenes as non-invasive biomarkers for ccRCC.Finally,we conducted on-line sample analysis of the selected LncRNAs and pseudogenes to determine their significant correlation with prognosis,and we try to investigate the possibility of them as a non-invasive biomarker for renal cancer prognosis and its possible mechanism affecting the development of ccRCC.Results:The selected eight LncRNAs and ten pseudogenes’ expression level was preliminarily tested using the blood samples from patients with ccRCC and healthy adults.The results showed that the changes in the expression level of pseudogenes PLEKHA8P1,CXCR2P1,KLRAP1,PDXDC2 P,FER1L4 and LncRNAs TLX1 NB,LINC00482 and DIO3 OS were relatively consistent and significant.Subsequently,a large cluster of samples were varified via real time PCR.The results showed significantly increased expression of pseudogene CXCR2P1(RQ=2.017,p=0.016),KLRAP1(RQ=3.152,p=0.010),PLEKHA8P1(RQ=2.782,p=0.014),PDXDC2P(RQ=2.625,p=0.029)and LncRNA TLX1NB(RQ=1.904,p=0.013)in the plasma of patients with ccRCC was significant,and the difference was statistically significant.Finally,by bioinformatics analysis,we found that the pseudogenes CXCR2P1,KLRAP1,PLEKHA8P1,and LncRNA TLX1 NB are associated with prognosis of patients with ccRCC.Conclusion:1.The system for extracting LncRNAs and pseudogenes from plasma and testing their expression level using real-time fluorescence quantitative PCR was successfully set up and verified.2.Pseudogenes CXCR2P1,KLRAP1,PLEKHA8P1 and PDXDC2 P and LncRNA TLX1 NB in plasma may be diagnostic non-invasive biomarkers of ccRCC.3.Pseudogenes CXCR2P1,KLRAP1 and PLEKHA8P1 and LncRNA TLX1 NB in plasma may be non-invasive biomarkers for the prognosis of ccRCC.