To clarify the mechanism of circGOLPH3 regulation on prostate cancer cells, we performed an over-expression 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 over-expression 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 over-expression 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 this study that circGOLPH3 and its binding protein CBX7can promote prostate cancer cell proliferation and inhibit apoptosis.

中文翻译：

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circGOLPH3及其结合蛋白CBX7对前列腺癌细胞增殖和凋亡的调控


为了阐明circGOLPH3对前列腺癌细胞的调控机制，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评估了前列腺癌细胞的细胞活力、增殖、细胞周期和凋亡。 、Edu 染色、TUNEL 染色和流式细胞术。通过 RNA Pull-down、质谱和 RNA 结合蛋白免疫沉淀 (RIP) 测定来鉴定 CircGOLPH3 的结合蛋白。通过qRT-PCR测量CircGOLPH3和CBX7的表达。结果显示，过表达circGOLPH3后，PC-3细胞的增殖能力和活力显着提高，而细胞凋亡受到抑制。 CircGOLPH3可以与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过表达显着提高了PC-3细胞的增殖能力和活力，并减少了细胞凋亡，这与circGOLPH3的作用一致。本研究验证了circGOLPH3及其结合蛋白CBX7能够促进前列腺癌细胞增殖并抑制细胞凋亡。