The Functional Study Of Long Non-coding RNA CRNDE In Bladder Cancer

BackgroundBladder cancer is the most common malignant tumor in urinary system,bringing a huge financial burden to society and threatening human health.During the past decades,great progress has been made in the diagnosis and therapy of bladder cancer.However,many problems concerning the treatment of bladder cancer have not been settled,such as undesirable side effects,lacking of accurate therapeutic target,low five-year survival rate and high rate of metastasis and recurrence.Surgery and intravesical instillation therapy are fit for bladder cancer patients in early stage.Palliative care is suitable for those in advanced stage,but the lethality rate of those patients remains high.Therefore,it is urgent to detect novel non-invasive biomarkers and accurate therapeutic targets for bladder cancer.Long non-coding RNAs are defined as RNA transcripts whose lengths are more than 200 nt,but do not have protein-coding ability.Recently,it is suggested that long non-coding RNAs play indispensable role in the initiation and development of cancers,and they are closely correlated with tumor metastasis and the prognosis of cancer patients.Among so many long non-coding RNAs,we focused on long non-coding RNA CRNDE.CRNDE is dysregulated in many kinds of tumors,promoting the proliferation,migration and invasion of cancer cells and acting as a cancer-promoting gene in the development of cancers.A new study indicates that high expression of CRNDE is closely correlated with the lymph nodes metastasis and TNM stage of cancer patients.Moreover,high expression of CRNDE leads to poor prognosis of cancer patients and it may serve as an indicator of the over survival of cancer patients.ObjectiveThis study aimed to investigate the expression level of CRNDE in the bladder cancer tissues and cell lines.The relationship between CRNDE expression and clinicopathological features would be analyzed.Moreover,we also detected the effects of CRNDE on the malignant phenotype of bladder cancer cells.MethodsTotal RNA from the bladder cancer tissues and matched tumor-adjcent tissues were extracted by using Trizol reagent.Quantitative Real-time PCR was performed to detect the expression level of CRNDE in the bladder cancer tissues and cell lines.In addition,chi-square test was used to analyze the correlation between the expression of CRNDE and the clinicopathological features of bladder cancer patients.In order to investigate the effects of CRNDE on the malignant phenotype of bladder cancer cells,we used the siRNA to inhibit CRNDE expression and transfected the overexpressed vector to increase CRNDE expression,respectively.Quantitative Real-time PCR was carried out to detect the validity of siRNA and overexpressed vector.Wound healing assay and Transwell assay were performed to measure the migration ability of bladder cancer cells.CCK-8 assay and EDU assay were carried out to detect the proliferation ability of bladder cancer cells.At last,ELISA capase-3 assay and flow cytometry were performed to detect the apoptosis of bladder cancer cells.ResultsLong non-coding RNA CRNDE is overexpressed in 54 bladder cancer tissues and cancer cell lines.High expression of CRNDE is positively associated with the TNM stage of bladder cancer patients,but irrelevant with the gender,age,histological grade,tumor size and lymph nodes metastasis of bladder cancer patients.In addition,CRNDE is overexpressed in bladder cancer cells(5637 and T24)compared with SV-HUC1 cell.Knockdown of CRNDE can inhibit cell migration and proliferation,while promote the apoptosis of bladder cancer cells.Overexpression of CRNDE can promote cell migration and proliferation,while inhibit the apoptosis of bladder cancer cells.ConclusionLong non-coding RNA CRNDE is augmented in bladder cancer tissues and cell lines,and the expression level of CRNDE is associated with the TNM stage.Long non-coding RNA CRNDE is involved in the development of bladder cancer and it can promote the migration and proliferation of bladder cancer cells,while inhibit cell apoptosis.To sum up,long non-coding RNA CRNDE acts as an oncogene in the development of bladder cancer,and it may become a novel diagnostic and therapeutic target for bladder cancer,providing theoretical basis for bladder cancer precise treatment in the future.