竞争性内源RNA，ncRAN，通过miR-1273e调节CDK6、RAP1A表达促进膀胱癌细胞增殖、侵袭机制的研究

The competing endogenous RNAs (ceRNAs) play an important role in the transcriptional regulation of the tumor transcriptome, but their roles in the malignant development of urinary tract tumors is unclear. We previously identified and analyzed a long non-coding RNA, non-coding RNA expressed in aggressive neuroblastoma (ncRAN), that promoted the development of urinary tract tumors. Further investigations showed that ncRAN shared miRNA-response elements with oncogenic transcripts of CDK6 and RAP1A, and promoted malignant progression of bladder cancer via regulation of CDK6 and RAP1A expressions. On the basis of these data, we hypothesized that ncRAN as a competing endogenous RNA for miR-1273e may promote bladder cancer proliferation and invasion via regulating CDK6 and RAP1A. In this project, we intends to obtain reliable evidences that ncRAN regulates CDK6 and RAP1A expressions via the miRNA pathway with a series of molecular biology methods such as luciferase reporter assay, RNA immunoprecipitation, and DNA microarray, combining with in vivo and in vitro functional experiments. The project will help to clarify transcriptional regulation mechanisms of the long non-coding RNAs, expand ceRNA regulatory networks and members, and provide a more solid scientific basis for the identification of new therapeutic targets for bladder cancer with the core characters of the ceRNA regulatory networks.

竞争性内源RNA(ceRNA)在转录组调控中发挥了重要作用,但其在泌尿系肿瘤恶性发展中的作用尚不清楚。申请者在前期工作中鉴定了一个促进泌尿系肿瘤恶性进展的长链非编码 RNA-ncRAN。进而发现ncRAN 和 癌基因CDK6、RAP1A的转录物通过竞争结合共享的miRNA应答元件调控各自的表达水平，影响膀胱癌的恶性进展。由此我们提出假说：ncRAN作为ceRNA，可通过miR-1273e调节CDK6、RAP1A的表达 ,促进膀胱癌细胞的增殖 、侵袭。本研究拟进一步采用荧光素酶报告基因分析、RNA蛋白免疫沉淀、基因芯片等分子生物学方法，结合体内、体外功能实验，旨在获得ncRAN经由miRNA途径调控 CDK6、RAP1A表达的可靠证据。本项目将有助于阐明长链非编码 RNA 的转录后调控机制，拓展ceRNA调控网络及成员,为鉴定以ceRNA调控网络为核心的膀胱癌治疗新靶点提供更充分的科学依据。

竞争性内源RNA(ceRNA)在转录组调控中发挥了重要作用,但其在泌尿系肿瘤恶性发展中的作用尚不清楚。申请者在前期工作中鉴定了一个促进泌尿系肿瘤恶性进展的长链非编码 RNA-ncRAN。进而发现ncRAN 和 癌基因CDK6、RAP1A 的转录物通过竞争结合共享的miRNA 应答元件调控各自的表达水平，影响膀胱癌的恶性进展。由此我们提出假说：ncRAN 作为ceRNA，可通过miR-1273e 调节CDK6、RAP1A 的表达 ,促进膀胱癌细胞的增殖 、侵袭。本研究拟进一步采用荧光素酶报告基因分析、RNA 蛋白免疫沉淀、基因芯片等分子生物学方法，结合体内、体外功能实验，旨在获得ncRAN 经由miRNA 途径调控 CDK6、RAP1A 表达的可靠证据。本项目将有助于阐明长链非编码 RNA 的转录后调控机制，拓展ceRNA 调控网络及成员,为鉴定以ceRNA 调控网络为核心的膀胱癌治疗新靶点提供更充分的科学依据。