Chengdu Second People's Hospital, Chengdu 10017, P.R. China.
Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, P.R. China.
Biosci Rep. 2020 Dec 23;40(12). doi: 10.1042/BSR20200936.
To clarify the mechanism of circGOLPH3 regulation on prostate cancer cells, we performed an overexpression and interference circGOLPH3 assay in prostate cancer cells PC-3 and then evaluated cellular viability, proliferation, cell cycle, and apoptosis of prostate cancer cells by MTT, CCK8, Edu stain, TUNEL stain, and flow cytometry. Binding proteins of CircGOLPH3 were identified by RNA pull-down, mass spectrometry, and RNA-binding protein immunoprecipitation (RIP) assays. The expressions of CircGOLPH3 and CBX7 were measured by qRT-PCR. The results showed that after overexpression of circGOLPH3, the proliferative capacity and the viability of PC-3cells were significantly improved, whereas apoptosis was inhibited. CircGOLPH3 could bind to the CBX7 protein that was highly expressed in the PC-3 cell. Additionally, a functional test on CBX7 showed that the CBX7 overexpression notably improved the proliferative capacity and the viability of PC-3 cells and decreased cellular apoptosis, which was consistent with the effects of circGOLPH3. The validated the present study that circGOLPH3 and its binding protein CBX7 can promote prostate cancer cell proliferation and inhibit apoptosis.
为了阐明 circGOLPH3 对前列腺癌细胞调控的机制,我们在前列腺癌细胞 PC-3 中进行了 circGOLPH3 的过表达和干扰实验,然后通过 MTT、CCK8、Edu 染色、TUNEL 染色和流式细胞术评估了前列腺癌细胞的细胞活力、增殖、细胞周期和凋亡。通过 RNA 下拉、质谱和 RNA 结合蛋白免疫沉淀(RIP)实验鉴定 CircGOLPH3 的结合蛋白。通过 qRT-PCR 测量 CircGOLPH3 和 CBX7 的表达。结果表明,circGOLPH3 过表达后,PC-3 细胞的增殖能力和活力显著提高,而凋亡受到抑制。CircGOLPH3 可以与在 PC-3 细胞中高表达的 CBX7 蛋白结合。此外,对 CBX7 的功能测试表明,CBX7 的过表达显著提高了 PC-3 细胞的增殖能力和活力,并降低了细胞凋亡,这与 circGOLPH3 的作用一致。验证了本研究表明 circGOLPH3 及其结合蛋白 CBX7 可以促进前列腺癌细胞的增殖并抑制凋亡。