Department of Urology, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, No.1 Shuaifuyuan Wangfujing, Dongcheng, Beijing, 100730, China.
J Cancer Res Clin Oncol. 2021 Mar;147(3):703-712. doi: 10.1007/s00432-020-03468-4. Epub 2021 Jan 1.
The malignant transformation of normal bladder cells (SV-HUC-1) was induced by arsenite to explore the possible mechanism of circRNA-100284 influencing bladder cancer cell proliferation.
Normal bladder SV-HUC-1 cells were cultured with 2 μM arsenite to induce malignant transformation. After 0, 3, 6, 12, and 24 h of culture, the expression level of circRNA-100284 in cells was detected by quantitative real-time PCR. Western blotting assays were used to detect the expression levels of EZH2 and cyclin-D1 proteins in cells treated with different media. Cell cycle was analyzed by flow cytometry. In addition, through cell transfection and CCK-8 experiments, the effect and mechanism of circRNA-100284 targeting microRNA-217 on proliferation was determined. The interaction between HSP70 methylation and Aurora-B was determined by Western blotting and immunoprecipitation experiments.
With prolonged contact time with arsenite, the expression level of circRNA-100284 in cells increased continuously (P < 0.05). Western blotting assays showed that the expression levels of EZH2 and cyclin-D1 proteins in arsenite-transformed cells increased. Flow cytometry and CCK-8 showed that circRNA-100284 accelerated cell cycle transition and cell proliferation through miR-217. Finally, after culturing human bladder cancer T24 cells, combined with immunoprecipitation and in vitro kinase experiments, it was found that K561- dimethyl HSP70 activated Aurora-B, thus promoting the proliferation of bladder cancer cells.
CircRNA-100284 activates aurora kinase B by inducing methylation of HSP70 via microRNA-217 to promote the proliferation of bladder cancer cells.
用亚砷酸钠诱导正常膀胱细胞（SV-HUC-1）恶性转化，探讨 circRNA-100284 影响膀胱癌细胞增殖的可能机制。
用 2 μM 亚砷酸钠培养正常膀胱 SV-HUC-1 细胞，诱导恶性转化。培养 0、3、6、12 和 24 h 后，采用实时定量 PCR 检测细胞中 circRNA-100284 的表达水平。采用 Western blot 检测不同培养基处理细胞中 EZH2 和细胞周期蛋白 D1 蛋白的表达水平。采用流式细胞术分析细胞周期。此外，通过细胞转染和 CCK-8 实验，确定 circRNA-100284 靶向 microRNA-217 对增殖的影响及其机制。通过 Western blot 和免疫沉淀实验确定 HSP70 甲基化与 Aurora-B 的相互作用。
随着与亚砷酸钠接触时间的延长，细胞中 circRNA-100284 的表达水平持续增加（P<0.05）。Western blot 检测结果显示，亚砷酸钠转化细胞中 EZH2 和细胞周期蛋白 D1 蛋白的表达水平升高。流式细胞术和 CCK-8 结果表明，circRNA-100284 通过 miR-217 加速细胞周期转换和细胞增殖。最后，在培养人膀胱癌 T24 细胞后，结合免疫沉淀和体外激酶实验发现，K561-二甲基 HSP70 激活 Aurora-B，从而促进膀胱癌细胞的增殖。
circRNA-100284 通过 microRNA-217 诱导 HSP70 甲基化激活 Aurora 激酶 B，促进膀胱癌细胞的增殖。
