| Objective:Bladder cancer(BCa)has a high incidence rate and tends to relapse.In most cases,the low 5-year survival rate in patients with advanced BCa is caused by postoperative recurrence and drug resistance.Long non-coding RNA(lnc RNA)can participate in many biological functions by regulating the expression of genes that affect tumorigenesis.A novel lnc RNA LINC02321 is associated with bladder cancer prognosis.The effect of LINC02321 on tumors is unknown.The research objective is to elaborate the important role of LINC02321/RUVBL2 signaling axis in bladder cancer progression,develop the clinical value of LINC02321 as new biomarkers in predicting tumor progression and intervening in tumor treatment,and provide a reference for the development of targeted drugs for bladder cancer.Methods:1.Bioinformatics was used to analyze the expression level of LINC02321 in pancarcinoma,and to evaluate the relationship between the expression level of LINC02321 and the survival prognosis of patients with bladder cancer.2.The expression level of LINC02321 in bladder cancer tissues and bladder cancer cell lines were determined by using q RT-PCR.3.The expression of LNC02321 and RUVBL2 was up-or down-regulated by lentiviral vector infection technique and transfection technique in bladder cancer cells.4.By using CCK-8,clone formation assay and flow cytometry,we analyzed the effects of LNC02321 and RUVBL2 expression on bladder cancer cell proliferation,clone formation ability,cell cycle and cisplatin sensitivity,respectively.5.RNA pulldown and mass spectrometry confirmed the interaction between LINC02321 and RUVBL2.The expression level of RUVBL2 protein and the half-life of the protein were verified by western blot.6.The tumorigenic experiments in nude mice confirmed that the LINC02321/RUVBL2 axis regulates the growth of bladder cancer cells in vivo and the sensitivity of bladder cancer cells to cisplatin.Results:1.LINC02321 is highly expressed in bladder cancer and is positively associated with shorter overall survival and disease-free survival in bladder cancer patients.LINC02321 differentially expressed genes in the high and low expression groups,enriched in tumor-related pathways.LINC02321 expression levels were high in the bladder cancer cell lines UMUC 3 and RT112,and LINC02321 were mainly distributed in the nucleus.2.Knocked down LINC02321 in UMUC3 and RT112,the proliferation and colony formation ability of the cells was inhibited,and the cell cycle was arrested in G0/G1 phase.And the sensitivity of bladder cancer cells to cisplatin was increased after LINC02321 knockdown.3.LINC02321 binds directly to RUVBL2 protein in bladder cancer cells.Knockdown and overexpression of LINC02321 did not affect RUVBL2 m RNA levels,but down-regulated or upregulated RUVBL2 protein expression levels.Knockdown of LINC02321 resulted in a shortened RUVBL2 protein half-life.RUVBL2 is highly expressed in bladder cancer and is positively associated with poor overall survival and disease-free survival in patients.4.In UMUC3 and RT112 cells,overexpression of RUVBL2 on the basis of LINC02321 knockdown partly restored the inhibition of cell proliferation and cell cycle caused by LINC02321 downregulation and suppressed the enhanced sensitivity to cisplatin caused by LINC02321 downregulation.5.In a nude mouse model,the carcinogenesis volume and mass of bladder cancer cell with LINC02321 knockdown were significantly reduced compared to control cells,and LINC02321 knockdown enhanced the sensitivity of bladder cancer cells to cisplatin in vivo.Conclusion:We demonstrated that the LINC02321/RUVBL2 signaling axis plays an important role in bladder cancer proliferation and cell sensitivity to cisplatin.RUVBL2 may be involved in the sensitivity of cisplatin in bladder cancer cells by regulating DNA repair.Cisplatin-resistant bladder cancer patients may benefit from RUVBL2 inhibitors combined with chemotherapy.LINC02321 is an oncogene in bladder cancer,is a potential target for evaluating tumor prognosis and interventioning in cancer therapy. |
PDF Full Text Request