An Angiogenesis-related LncRNA Signature For The Prognostic Prediction Of Patients With Bladder Cancer And LINC02321 Promotes Bladder Cancer Progression Via The VEGFA Signaling Pathway

Objective:To constructs a angiogenesis related IncRNA prognosis model through big data mining,explores the relationship between this model and the survival prognosis of bladder cancer patients,and explores the role and mechanism of angiogenesis related long chain non coding RNA LINC02321 in the progression of bladder cancer.Methods:Download transcriptome data of bladder cancer and adjacent tissue samples and clinical data of bladder cancer patients in TCGA database.At the same time,download the gene set related to angiogenesis in the molecular model database.Pearson correlation analysis was used to screen angiogenesis related lncRNA,and Cox regression analysis was used to construct a angiogenesis related lncRNA model related to the prognosis of bladder cancer patients.According to the model,we screened the angiogenesis related lncRNA:LINC02321 that probably interfere with the proliferation and migration of bladder cancer.After determining the research objectives,the expression of LINC02321 was verified by RT qPCR at the tissue level and cell level respectively;After lentivirus overexpression of LINC02321 and small interfering RNA knockdown of LINC02321,CCK-8 cell viability test was used to detect the effect of LINC02321 on the viability and proliferation of bladder cancer cells;The effect ofLINC02321 on migration and invasion of bladder cancer cells was tested by Transwell cell migration,invasion test and cell scratch test.The effect of LINC02321 on cisplatin drug sensitivity was detected by CCK-8 drug sensitivity test;We further explored the mechanism of LINC02321 in the malignant progression of bladder cancer through GSEA enrichment analysis,Pearson correlation analysis and Western blot experiment.Results:In the TCGA database,we found 390 angiogenesis related lncRNAs through Pearson correlation analysis of lncRNA transcription data and angiogenesis gene expression data of bladder cancer samples.Combined with the clinical data of bladder cancer patients,COX regression analysis was carried out on the 390 angiogenesis related lncRNAs,and a 6-angiogenesis related lncRNA prediction model(USP30-AS1,LINC02321,PSMB8-AS1,KRT7-AS,LINC01767 and OCIAD1-AS1)was successfully constructed.According to the median of risk score(RS),patients were divided into high-risk and low-risk groups.The study showed that the total survival time(OS)of patients in high-risk group was significantly shorter than those in low-risk group,and the highest AUC value of receiver operating characteristic curve(ROC)curve was 0.807.The nomograph is constructed based on the patient’s age,sex,stage and model risk score.Compared with traditional clinical indicators,the model risk score we have constructed has a higher predictive efficacy for the patient’s prognosis.By drawing the calibration curve and ROC curve,it is found that the accuracy of the nomograph is higher.TCGA database prediction and RT qPCR experiment results showed that only LINC02321 was highly expressed in bladder cancer tissue,And accelerate the replication,metastasis and cisplatin resistance of malignant tumors.GSEA,Pearson analysis and Western blot experiments showed that VEGFA was involved in LINC02321 regulated bladder cancer progression.Conclusion:The LINC02321 is significantly up-regulated in bladder cancer tissues and cell lines,which may promote the proliferation and metastasis of bladder cancer cells by inducing the high expression of VEGFA,and increase the resistance of bladder cancer cells to cisplatin,thus having a adverse effect on the prognosis of bladder cancer patients.