Expression And Function Study Of Circular RNA ZKSCAN1 In Bladder Cancers

BACKGROUND AND OBJECTIVE: Bladder malignancy is the most common tumor in the urinary system.The incidence of bladder cancer is high,and the mortality rate is also high.It imposes a heavy burden on society and individuals.The diagnosis and treatment of bladder cancer has made great progress,its 5-year survival rate is still unsatisfactory.The treatments for bladder cancer include surgery,radiotherapy and chemotherapy,and biological therapy.These treatments have a serious impact on the patient’s quality of life.Finding molecular markers that influence the development of bladder cancer and understanding the underlying pathogenesis of bladder cancer will be of great significance in the early diagnosis and treatment of bladder cancer.Circular RNA is a research hotspot in recent years.It has been found that many circular RNAs are abnormally expressed in tumor tissues and have important influence on tumor progression.Circ-ZKSCAN1 is a recently discovered circular RNA.Studies have shown that it has an inhibitory effect on the development of hepatocellular carcinoma.However,it has not been reported in bladder cancer.Therefore,this study aims to study the expression of Circ-ZKSCAN1 in bladder cancer and verify its effect on the function of bladder cancer cells.In order to further regulate the expression of Circ-ZKSCAN1 in bladder cancer,we constructed a target genes quantitative regulation system for bladder cancer according to the principles of synthetic biology.We also verify its feasibility.METHODS: We used QRT-PCR to detect the relative expression levels of circ-ZKSCAN1 in 51 pairs of bladder cancer tissue specimens and two bladder cancer cell lines.Then we also analyzed the relationship between the expression level of circ-ZKSCAN1 and clinical pathological features.We constructed a circ-ZKSCAN1 overexpression plasmid to up-regulate the expression level of circ-ZKSCAN1 in bladder cancer cell lines,and then verified its effects on the proliferation,migration and invasion of bladder cancer cells.We inserted a guide RNA that binds to the GFP gene in a bladder cancer target gene quantitative regulatory system to construct a verification plasmid.We used 293 T cells that stably expressing the GFP gene to verify the feasibility of this synthetic system.RESULTS: The expression level of circ-ZKSCAN1 in bladder cancer tissues was significantly lower than that of paired adjacent normal tissues.The expression level of circ-ZKSCAN1 was related to the tumor’s primary tumor.In the bladder cancer cell line,the expression level of circ-ZKSCAN1 is also significantly lower than normal bladder urothelial cells.Circ-ZKSCAN1 overexpression plasmid can effectively increase the expression level of circ-ZKSCAN1 in bladder cancer cells.In addition,transfection of Circ-ZKSCAN1 overexpression plasmid into bladder cancer cells can significantly inhibit the proliferation,migration and invasion of the cells.The expression level of GFP gene was significantly inhibited after the target gene quantitative regulation system of the bladder cancer was transferred to the 293 T cells that stably expressing the GFP gene.CONCLUSION: As a tumor suppressor gene,Circ-ZKSCAN1 plays an important role in the progression of bladder cancer and may be a novel molecular biomarker in bladder cancer.Bladder cancer target gene quantitative regulation system may become a new method to study bladder cancer and regulate gene expression.