Department of Urology, the First Affiliated Hospital of Xiamen University, No.55 Zhenhai Road, Xiamen, 361003, Fujian, China.
Mol Med. 2018 Jul 31;24(1):40. doi: 10.1186/s10020-018-0039-0.
This study aimed to investigate the effect of over-expressing circular RNA CEP128 (circCEP128) on cell functions and explore the molecular mechanism of which in bladder carcinoma.
The differentially expressed circRNAs and mRNAs in bladder carcinoma cells and cells in adjacent tissues were screened out using microarray analysis. Expression levels of circRNAs and mRNAs in tissues and cells were determined by qRT-PCR. Expression of SOX11 was detected by western blot. Luciferase reporter assay and RNA pull-down assay were used to investigate the interactions between the specific circRNA, miRNA and mRNA. Cell cycle and apoptosis were measured using flow cytometry after transfection. MTT assay was also performed to detect the cell proliferation.
In present study, circCEP128 and SOX11 were observed significantly up-regulated in bladder cancer tissues, while the expression of miR-145-5p was decreased in cancer samples compared to normal samples. Cytoscape was used to visualize circCEP128-miRNA-target gene interactions based on the TargetScan and circular RNA interactome, which revealed that circCEP128 served as a sponge of miR-145-5p and indirectly regulated SOX11. Knockdown of circCEP128 induced the inhibition of cell proliferation and the increased bladder cancer cell apoptosis rate.
CircCEP128 functions as a ceRNA for miR-145-5p, which could up regulates SOX11 and further promotes cell proliferation and inhibits cell apoptosis of bladder cancer.
本研究旨在探讨过表达环状 RNA CEP128（circCEP128）对膀胱癌细胞功能的影响，并探索其在膀胱癌中的分子机制。
通过微阵列分析筛选出膀胱癌细胞和相邻组织细胞中差异表达的环状 RNA 和信使 RNA。采用 qRT-PCR 法检测组织和细胞中 circRNA 和 mRNAs 的表达水平。采用 Western blot 检测 SOX11 的表达。采用荧光素酶报告基因和 RNA 下拉实验检测特定 circRNA、miRNA 和 mRNA 之间的相互作用。转染后通过流式细胞术检测细胞周期和凋亡。采用 MTT 法检测细胞增殖。
本研究发现 circCEP128 和 SOX11 在膀胱癌组织中明显上调，而 miR-145-5p 在癌症样本中的表达低于正常样本。Cytoscape 软件基于 TargetScan 和环状 RNA 相互作用网络可视化 circCEP128-miRNA-靶基因相互作用，表明 circCEP128 作为 miR-145-5p 的海绵分子，间接调控 SOX11。circCEP128 敲低可诱导膀胱癌细胞增殖抑制和细胞凋亡率增加。
circCEP128 作为 miR-145-5p 的 ceRNA，可上调 SOX11，进而促进膀胱癌的细胞增殖和抑制细胞凋亡。
