Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, No.1 Shuaifuyuan, Dongcheng District, Beijing, China.
Department of Gynecology and Obstetrics, West China Second University Hospital, Sichuan University, Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University), Ministry of Education, Chengdu, China.
J Ovarian Res. 2018 Apr 23;11(1):31. doi: 10.1186/s13048-018-0406-z.
B-cell-specific Moloney murine leukemia virus integration site 1 (BMI1) might be an appropriate biomarker in the management of epithelial ovarian cancer (EOC). However, the biological role of BMI1 and its relevant molecular mechanism needs further elaboration. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system is an excellent genome-editing tool and is scarcely used in EOC studies.
We first applied CRISPR/Cas9 technique to silence BMI1 in EOC cells; thereafter we accomplished various in vivo and in vitro experiments to detect biological behaviors of ovarian cancer cells, including MTT, flow cytometry, Transwell, real-time polymerase chain reaction and western blotting assays, etc.; eventually, we used RNA sequencing to reveal the underlying molecular traits driven by BMI1 in EOC.
We successfully shut off the expression of BMI1 in EOC cells using CRISPR/Cas9 system, providing an ideal cellular model for investigations of target gene. Silencing BMI1 could reduce cell growth and metastasis, promote cell apoptosis, and enhance the platinum sensitivity of EOC cells. BMI1 might alter extracellular matrix structure and angiogenesis of tumor cells through regulating Focal adhesion and PI3K/AKT pathways.
BMI1 is a potential biomarker in EOC management, especially for tumor progression and chemo-resistance. Molecular traits, including BMI1 and core genes in Focal adhesion and PI3K/AKT pathways, might be alternatives as therapeutic targets for EOC.
B 细胞特异性 Moloney 鼠白血病病毒整合位点 1（BMI1）可能是上皮性卵巢癌（EOC）管理的合适生物标志物。然而，BMI1 的生物学作用及其相关分子机制需要进一步阐述。成簇规律间隔短回文重复（CRISPR）/Cas9 系统是一种优秀的基因组编辑工具，在 EOC 研究中很少使用。
我们首先应用 CRISPR/Cas9 技术沉默 EOC 细胞中的 BMI1；此后，我们完成了各种体内和体外实验，以检测卵巢癌细胞的生物学行为，包括 MTT、流式细胞术、Transwell、实时聚合酶链反应和 Western blot 等；最终，我们使用 RNA 测序揭示 BMI1 在 EOC 中驱动的潜在分子特征。
我们成功地使用 CRISPR/Cas9 系统关闭了 EOC 细胞中 BMI1 的表达，为靶基因的研究提供了理想的细胞模型。沉默 BMI1 可降低细胞生长和转移，促进细胞凋亡，并增强 EOC 细胞对铂类的敏感性。BMI1 可能通过调节黏附斑和 PI3K/AKT 通路改变肿瘤细胞的细胞外基质结构和血管生成。
BMI1 是 EOC 管理的潜在生物标志物，特别是对肿瘤进展和化疗耐药性。包括 BMI1 在内的分子特征以及黏附斑和 PI3K/AKT 通路中的核心基因，可能成为 EOC 的替代治疗靶点。
