Synthetic Circular RNA Elements Inhibit The Malignant Phenotype Of Bladder Cancer Cell 5637 By Targeting MicroRNA-183 Cluster And Transcription Factor β–Catenin

Purpose: Many studies have shown that transcription factor β-Catenin and mi RNA-183 cluster can play a regulatory role in the occurrence and development of tumor.Previous studies have constructed an artificial long non coding RNA,which can achieve the simultaneous regulation of transcription factor β-Catenin and mi RNA-183 cluster.However,linear non coding RNA is unstable in cells.We want to construct a more stable synthetic biological element that can regulate transcription factors and mi RNA at the same time.Circular RNA is stable,and it can be used as a molecular sponge of mi RNA and RNA aptamer.In addition,the technology of in vitro synthesis of circular RNA has become mature.Therefore,our goal is to construct a synthetic biological element,which can form circular RNA in cells,and the formed circular RNA sequence is an artificially designed target sequence.We call this element acirc RNA,which can target the transcription factors β-Catenin and mi RNA-183 cluster in tumor cells,thus achieving the regulation of tumor cells.Methods: In order to realize the function of circular RNA element,we used the principle of synthetic biology to arrange two β-Catenin aptamer c DNA sequences and four mi RNA-183-182-96 cluster coordination point c DNA sequences in series,and inserted them into the filling sequence of artificial overexpression vector of circular RNA,so as to construct the acirc RNA we need.The acirc RNA element was transfected into 5637 cells.The regulatory effect of acirc RNA element on target molecules and phenotype of 5637 cells were verified by fluorescence quantitative PCR,CCK-8,edu and flow cytometry.In addition,we constructed four plasmids to verify whether the element can form a loop in cells.Results: The four plasmids were transferred into 293 t cells,and the fluorescence expression of each group was consistent with the expected results,indicating that the circular RNA element constructed by us could form circular RNA in cells.PCR results showed that the circular RNA element could reduce the expression of mi RNA-183-182-96 cluster and β-Catenin.Further functional experiments showed that the circular RNA element could affect the proliferation,migration and apoptosis of 5637 cells.Conclusion: The circular RNA elements synthesized by us can form specific circular RNA molecules in cells and play a role in inhibiting tumor cell phenotype,and achieve simultaneous pre-and post transcriptional regulation in bladder cancer cell 5637.However,further research still needs to be improved,and there is still a certain distance to apply it to the treatment of tumor.