LncRNA-CP Affects Invasion And Migration Of Renal Clear Cell Carcinoma By Regulating HIF-1α/VEGFA Axis

Objective: Renal Cell Carcinoma(RCC)is one of the most common malignant tumors of the urinary system in adults,and clear Cell Carcinoma of the kidney(cc RCC)is the most common subtype of Renal cancer.Due to the complex nature of renal clear cell carcinoma(RCC),its mechanism has not been fully understood.Long non-coding RNA,as a potential biomarker of various cancers,has become a research hotspot,but its research in renal clear cell carcinoma is still largely unknown.Therefore,this paper aims to search for a new long non-coding RNA in renal clear cell carcinoma,and to explore the mechanism of its potential as a new biomarker and therapeutic target for renal clear cell carcinoma.This article first identified lnc RNA(which we named lnc RNA-CP),which is highly expressed in renal clear cell carcinoma and has prognostic value from TCGA.We confirmed its locus information in the UCSC database,and found that the gene CP was adjacent to its genome,and then we discussed whether there was an expression correlation between the two and predicted the cis-regulatory mechanism between them.We have also found that CP is a known target of HIF-1α and VEGFA in vitro,and transcription-induced CP promoter activation in hypoxic environment leads to increased expression of CP.Therefore,we focus on whether lnc RNA-CP affects the invasion and migration of renal clear cell carcinoma by regulating the HIF-1α/VEGFA axis.Methods: We firstly screened and identified new lnc RNA,lnc RNA-CP from the TCGA database.Differential expression analysis was performed to observe the expression of lnc RNA-CP in cc RCC,and KM-plot was used to analyze the relationship between the expression of lnc RNA-CP and the prognosis of cc RCC.The cross table was used to analyze the relationship between the expression of lnc RNA-CP and the clinicopathological parameters of cc RCC,and Cox proportional risk regression model was used to analyze whether lnc RNA-CP could be used as an independent prognostic risk factor of cc RCC.UCSC database was used to determine the proximity between the loci of lnc RNA-CP and CP.The expression and distribution of lnc RNA-CP in ACHN and 786-O cells of renal clear cell carcinoma were determined by cytoplasmic separation.m RNA and protein expression of related genes HIF-1α and VEGFA were detected by q RT-PCR and Western Blot.Meanwhile,cell scratch and Transwell experiments were performed to investigate the effect of silencing lnc RNA-CP on biological characteristics.Results: 1.Through the TCGA database,we identified 12 lnc RNAs with high prognostic value in cc RCC and single-factor and multi-factor analysis of lnc RNA,so we can obtain three lnc RNAs as independent risk factors for cc RCC.They are RP11-206M11.7,RP5-884M6.1 and LINC00460,respectively.Finally,we also analyzed the clinical pathology of these genes and found that they are related to TNM stage or Fuhrman(grade).The influence of RP11-206M11.7 in renal clear cell carcinoma has not been reported.We next select RP11-206M11.7 for the next analysis.(We named it lnc RNA-CP)2.It was verified by nuclear separation experiments that lnc RNA-CP was mainly localized in the nucleus.The positional relationship between lnc RNA-CP and CP loci was determined by USCS database.TCGA data showed that the expression of both was positively correlated;CP was highly expressed in renal clear cell carcinoma and was associated with poor prognosis.Clinical case parameter analysis of CP showed that it was related to gender,grade.3.Silent lnc RNA-CP cell model was successfully constructed.At the same time,q RT-PCR was used to detect the expression of CP m RNA.It was found that the expression of CP decreased in the same direction after silencing lnc RNA-CP.The scratching ability and transwell experiment showed that the cell migration ability was weakened after silencing lnc RNA-CP.After silencing lnc RNA-CP in 786-O and ACHN,the m RNA expression levels of VEGFA and HIF-1α were decreased by q RT-PCR.At the same time,western blot analysis showed that the expression levels of VEGFA and HIF-1α protein decreased in the same direction after interference with lnc RNA-CP,and there was no significant change in VEGF-R.Conclusion: 1.lnc RNA-CP is highly expressed in renal clear cell carcinoma and is an independent prognostic risk factor.2.The lnc RNA-CP in cc RCC was positively correlated with the expression of adjacent gene CP.3.Silencing lnc RNA-CP inhibits cc RCC invasion and migration.4.Silencing lnc RNA-CP inhibits HIF-1α and VEGFA expression in cc RCC.