Reproductive Medicine Center, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China.
Department of Intensive Care Unit, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China.
Mol Med Rep. 2021 Jul;24(1). doi: 10.3892/mmr.2021.12155. Epub 2021 May 20.
Ovarian cancer is one of the most common lethal gynecological malignancies worldwide. Abnormal kinesin family member 4A (KIF4A) expression has been implicated in ovarian cancer progression; however, the potential mechanism underlying KIF4A in ovarian cancer is not completely understood. The present study aimed to clarify the molecular basis of KIF4A in ovarian cancer. KIF4A and budding uninhibited by benzimidazoles 1 (BUB1) expression levels were detected via reverse transcription-quantitative PCR and western blotting. Cell Counting Kit-8, colony formation, wound healing, TUNEL and flow cytometry assays were performed to assess cell proliferation, migration, apoptosis and cell cycle distribution, respectively. Ki67 expression levels were detected by conducting immunofluorescence assays. The expression levels of migration- and apoptosis-related proteins were measured via western blotting. A co-immunoprecipitation assay was conducted to determine the association between KIF4A and BUB1. The results demonstrated that KIF4A was expressed at significantly higher levels in ovarian cancer cell lines compared with IOSE-80 cells. Compared with the short hairpin RNA-negative control group, KIF4A knockdown significantly inhibited cell viability, colony formation and migration, and markedly induced cell apoptosis. The results indicated that KIF4A could bind to BUB1 and regulate BUB1 expression. BUB1 overexpression weakened KIF4A knockdown-mediated effects on cell viability, colony formation, migration and apoptosis. Overall, the present study demonstrated that KIF4A knockdown suppressed ovarian cancer progression by regulating BUB1, and suggested the potential value of KIF4A and BUB1 as therapeutic targets for ovarian cancer.
卵巢癌是全球最常见的致命妇科恶性肿瘤之一。异常的驱动蛋白家族成员 4A（KIF4A）表达与卵巢癌的进展有关；然而，KIF4A 在卵巢癌中的潜在机制尚不完全清楚。本研究旨在阐明 KIF4A 在卵巢癌中的分子基础。通过逆转录定量 PCR 和 Western blot 检测 KIF4A 和有丝分裂检验点激酶 1（BUB1）的表达水平。通过细胞计数试剂盒-8、集落形成、划痕愈合、TUNEL 和流式细胞术分别评估细胞增殖、迁移、凋亡和细胞周期分布。通过免疫荧光法检测 Ki67 表达水平。通过 Western blot 测定迁移和凋亡相关蛋白的表达水平。进行免疫共沉淀测定以确定 KIF4A 与 BUB1 之间的关联。结果表明，与 IOSE-80 细胞相比，KIF4A 在卵巢癌细胞系中的表达水平明显更高。与短发夹 RNA 阴性对照组相比，KIF4A 敲低显著抑制细胞活力、集落形成和迁移，并显著诱导细胞凋亡。结果表明，KIF4A 可以与 BUB1 结合并调节 BUB1 的表达。BUB1 过表达削弱了 KIF4A 敲低介导的对细胞活力、集落形成、迁移和凋亡的影响。总体而言，本研究表明 KIF4A 敲低通过调节 BUB1 抑制卵巢癌进展，并提示 KIF4A 和 BUB1 作为卵巢癌治疗靶点的潜在价值。
