Department of Gynaecology and Obstetrics, Joint Service Support Force No.920 Hospital of the PLA, No. 212 Daguan Road, Xishan District, Kunming, 650032, Yunnan Province, China.
Mol Biotechnol. 2022 Sep;64(9):958-969. doi: 10.1007/s12033-022-00463-7. Epub 2022 Mar 9.
Cyclin B1 (CCNB1) is regarded as an oncogene in multiple tumors. This work aims to investigate the expression, function, and related mechanisms of CCNB1 in ovarian carcinoma (OC). Three microarray datasets (GSE14407, GSE18520, and GSE54388) were obtained from the Gene Expression Omnibus (GEO) database and screened for differentially expressed genes (DEGs) of OC tissues and normal ovarian tissues. CCNB1 expression in OC tissues and paracancerous tissues was detected by immunohistochemistry. Kaplan-Meier plotter database was utilized to analyze the correlation between CCNB1 expression and the prognosis of OC patients. After the loss-of-function and gain-of-function cell models were established, cell counting kit-8 (CCK-8), bromo-deoxyuridine (BrdU), and transwell experiments were employed to examine the proliferation, migration, and invasion of OC cells, respectively. The targeting relationship between miR-559 and CCNB1 was verified using the dual-luciferase reporter gene experiment. The expressions of CCNB1 mRNA and miR-559 were detected by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Western blot was used to quantify the protein expression of CCNB1. In addition, xenograft nude mouse models were established to examine the effects of CCNB1 on lung metastasis in vivo. CCNB1 expression was markedly increased in OC tissues and cell lines. The overall survival, progression-free survival, and post-progression survival of OC patients with high CCNB1 expression were significantly shorter. OC cell proliferation, migration, and invasion were enhanced by CCNB1 overexpression while CCNB1 knockdown led to opposite effects. MiR-559 expression was remarkably reduced in OC tissues and cell lines, and miR-559 markedly suppressed the malignant characteristics of OC cells. Besides, miR-559 directly targeted the 3' UTR of CCNB1 mRNA and reduced CCNB1 expression at both the mRNA and protein levels. Overexpression of CCNB1 accelerated lung metastasis of OC cells in vivo. CCNB1, of which expression is modulated by miR-559, facilitates proliferation, migration, and invasion of OC cells, therefore, working as a potential therapeutic target of OC. This work provides new insights into the clinical diagnosis and treatment of OC.
细胞周期蛋白 B1 (CCNB1) 被认为是多种肿瘤的癌基因。本研究旨在探讨 CCNB1 在卵巢癌 (OC) 中的表达、功能及相关机制。从基因表达综合数据库 (GEO) 中获取了 3 个微阵列数据集 (GSE14407、GSE18520 和 GSE54388)，筛选 OC 组织和正常卵巢组织差异表达基因 (DEGs)。采用免疫组织化学法检测 OC 组织和癌旁组织中 CCNB1 的表达。利用 Kaplan-Meier plotter 数据库分析 CCNB1 表达与 OC 患者预后的相关性。建立 CCNB1 敲低和过表达细胞模型后，通过细胞计数试剂盒-8 (CCK-8)、溴脱氧尿苷 (BrdU) 和 Transwell 实验分别检测 OC 细胞的增殖、迁移和侵袭能力。通过双荧光素酶报告基因实验验证 miR-559 与 CCNB1 的靶向关系。采用实时定量逆转录聚合酶链反应 (qRT-PCR) 检测 CCNB1 mRNA 和 miR-559 的表达。采用 Western blot 法检测 CCNB1 蛋白表达。此外，建立 OC 裸鼠肺转移模型，检测 CCNB1 体内对肺转移的影响。CCNB1 在 OC 组织和细胞系中表达明显上调。CCNB1 高表达的 OC 患者总生存期、无进展生存期和进展后生存期明显缩短。CCNB1 过表达促进 OC 细胞增殖、迁移和侵袭，而 CCNB1 敲低则产生相反的效果。miR-559 在 OC 组织和细胞系中的表达明显下调，miR-559 显著抑制 OC 细胞的恶性特征。此外，miR-559 可直接靶向 CCNB1 mRNA 的 3'UTR，降低 CCNB1 mRNA 和蛋白水平的表达。CCNB1 过表达加速 OC 细胞体内肺转移。CCNB1 受 miR-559 调控，促进 OC 细胞增殖、迁移和侵袭，因此，CCNB1 可能成为 OC 的潜在治疗靶点。本研究为 OC 的临床诊断和治疗提供了新的思路。
