Role And Mechanism Of LINC00858 In Bladder Cancer

Bladder cancer is one of the most common cancers globally.Because bladder cancer is easy to relapse after treatment,the current mortality rate of patients with bladder cancer is high and the 5-year survival rate is still low.Therefore,it is necessary to further study the pathogenesis of bladder cancer,exploring new target for bladder cancer treatment is urgently needed to further improve the therapeutic effects on bladder cancer.The long non-coding RNA00858(LINC00858)has been reported to be an oncogene for various cancer diseases,including osteosarcoma and colorectal cancer.LINC00858 has been found to induce the progression and metastasis of malignant tumors,and has been studied as a biomarker and novel target for the diagnosis and treatment of a variety of tumors.However,the expression and function of LINC00858 in bladder cancer remain unclear.This project aims to study the role and mechanism of LINC00858 in the occurrence and development of bladder cancer,and is expected to provide theoretical basis for the development of new targets and drugs for the treatment of bladder cancer.Part1:The role of LINC00858 in the development of bladder cancerPurposes:To explore the expression of LINC00858 in bladder cancer tissues and cells,and to study the malignant biological function role of LINC00858 in bladder cancer.Methods:RT-qPCR was used to detect the expression of LINC00858 in bladder cancer tissues and normal adjacent tissues,as well as in bladder cancer cell lines(5637,T24 and J82)and bladder epithelial cell lines(SV-HUC-1).Nuclear and cytoplasmic separation experiments analysis the expression of LINC00858 in cytoplasm and nucleus of bladder cancer cells.Gene interference technology(knock down the expression of LINC00858 of bladder cancer cell),CCK-8 experiment,clone formation experiment,wound healing experiment and transwell chamber experiment were uesd to detect the changes of cell proliferation,migration and invasion.Results:The expression of LINC00858 in bladder cancer tissue was significantly higher than that in normal tissues adjacent to cancer(P<0.001).In addition,the expression of LINC00858 in bladder cancer cell lines was significantly higher than that in normal human bladder epithelial cell lines(P<0.01).LINC00858 is mainly expressed in the cytoplasm.Knockdown of LINC00858 of bladder cancer cells can significantly inhibit cell proliferation,migration and invasion.Conclusion:LINC00858 is highly expressed in bladder cancer and may mediate the malignant biological function of bladder cancer.Part2:The mechanism of LINC00858 in the development of bladder cancerPurposes:To explore the molecular mechanism of LINC00858 in regulating the development of bladder cancer.Methods:Bioinformatics analysis was performed by starbasev2.0 to predict the miRNA bound to LINC00855 and downstream target genes.Dual luciferase reporters and RNA immunoprecipitation(RIP)were used to identify the interaction between LINC00858,miR-3064-5p and CTGF(connective tissue growth factor).Gene interference and gene overexpression technology,CCK-8 experiment,clone formation experiment,wound healing experiment and transwell chamber experiment were used to verify the relationship between LINC00858 and miR-3064-5p/CTGFin bladder cancer cells.Results:LINC00858 directly targets miR-3064-5p in bladder cancer cells.Inhibition of miR-3064-5p promoted the proliferation,migration and invasion of bladder cancer cells.RIP experiment results showed that CTGF is the direct target of miR-3064-5p.miR-3064-5p mimics significantly reduced the expression of CTGF in bladder cancer cells,which could be restored by LINC00858 overexpression.Overexpression of LINC00858 significantly enhanced the proliferation,migration and invasion of bladder cancer cells.However,konockdown of CTGF gene can antagonize the above-mentioned biological behaviors.Conclusion:LINC00858,as a competitive RNA,mediates the malignant biological ability of bladder cancer by isolating miR-3064-5p to increase the expression level of oncogene CTGF.LINC00858/miR-3064-5p/CTGF axis might be an important pathway for the occurrence and development of bladder cancer.