Department of Obstetrics and Gynaecology, Renmin Hospital of Wuhan University, Jiefang Road No.238, Wuhan, 430060, Hubei, China.
Mol Biotechnol. 2021 Dec;63(12):1235-1243. doi: 10.1007/s12033-021-00375-y. Epub 2021 Aug 2.
Ovarian cancer (OC) is one of the most common cancers among women, characterized by various histological subtypes. Here, we aimed to investigate the biological function of F-box and leucine-rich repeat protein 20 (FBXL20) in the malignant phenotype of OC cells and its related mechanism. The expression of FBXL20 in OC tissue and normal tissue samples was analyzed through the GEPIA database. Quantitative real-time PCR (qRT-PCR), immunohistochemistry (IHC) and Western blot were employed to detect the expression of miR-195-5p and FBXL20 in OC tissues and cell lines. Cell counting kit-8 (CCK-8) assay, 5-ethynyl-2'-deoxyuridine (EdU) experiment and flow cytometry were applied to detect cell proliferation, cell cycle and apoptosis. Bioinformatics analysis and dual-luciferase reporter gene experiments were adopted to predict and validate the targeting relationship between miR-195-5p and FBXL20 mRNA 3'-untranslated region (3'UTR). Correlation between the expressions of miR-195-5p and FBXL20 mRNA was analyzed by Pearson correlation analysis. FBXL20 expression was upregulated in OC, and its high expression level was significantly associated with higher International Federation of Gynecology and Obstetrics (FIGO) stage and poor tumor differentiation. Functionally, overexpression of FBXL20 promoted proliferation, inhibited apoptosis and accelerated the cell cycle in OC cells in comparison to control group, and knockdown of FBXL20 exerted the opposite effects. Mechanistically, miR-195-5p directly targeted FBXL20 and negatively regulated its expression. Pearson correlation analysis indicated that miR-195-5p was negatively correlated with FBXL20 mRNA expression. In addition, overexpression of miR-195-5p reversed the above biological functions of FBXL20 in OC cells. FBXL20, negatively regulated by miR-195-5p, accelerates the proliferation and cell cycle progression of OC cells, and inhibits cell apoptosis, which might act as a prospective prognostic biomarker and a promising therapeutic target for OC.
卵巢癌 (OC) 是女性中最常见的癌症之一，其特征是存在各种组织学亚型。在这里，我们旨在研究 F-box 和富含亮氨酸重复蛋白 20 (FBXL20) 在 OC 细胞恶性表型中的生物学功能及其相关机制。通过 GEPIA 数据库分析 OC 组织和正常组织样本中 FBXL20 的表达。采用定量实时 PCR (qRT-PCR)、免疫组织化学 (IHC) 和 Western blot 检测 OC 组织和细胞系中 miR-195-5p 和 FBXL20 的表达。细胞计数试剂盒-8 (CCK-8) 检测、5-乙炔基-2'-脱氧尿苷 (EdU) 实验和流式细胞术用于检测细胞增殖、细胞周期和细胞凋亡。采用生物信息学分析和双荧光素酶报告基因实验预测和验证 miR-195-5p 和 FBXL20 mRNA 3'-非翻译区 (3'UTR) 的靶向关系。采用 Pearson 相关性分析分析 miR-195-5p 和 FBXL20 mRNA 表达之间的相关性。分析 FBXL20 在 OC 中的表达上调，其高表达水平与较高的国际妇产科联合会 (FIGO) 分期和较差的肿瘤分化显著相关。功能上，与对照组相比，FBXL20 的过表达促进 OC 细胞的增殖，抑制凋亡并加速细胞周期，而 FBXL20 的敲低则产生相反的效果。机制上，miR-195-5p 直接靶向 FBXL20 并负调控其表达。Pearson 相关性分析表明 miR-195-5p 与 FBXL20 mRNA 表达呈负相关。此外，miR-195-5p 的过表达逆转了 FBXL20 在 OC 细胞中的上述生物学功能。FBXL20 受 miR-195-5p 的负调控，加速 OC 细胞的增殖和细胞周期进程，抑制细胞凋亡，可能作为 OC 的一个有前途的预后生物标志物和有希望的治疗靶点。
