knockout of EZH2 inhibits the tumorigenicity of human ovarian cancers cells SKOV3 by targeting steroid biosynthesis

Ovarian cancer has the highest mortality in gynecologic cancers and most patients are diagnosed in advanced stages. EZH2 has been a major tumor markers and effective therapeutic targets for ovarian cancer, which molecular mechanism remain unclear. The present study was investigated the biological effects on EZH2 knock-out SKOV3 cells in vitro and in vivo experiment, and explore the molecular mechanism by integrated analysis of mRNA-seq and CHIP-seq.
 CRISPR/Cas9 system was used to establish the knock-out EZH2 SKOV3 cells. Protein expression was detected by Western blot. The effect of EZH2 in ovarian cancer was detected by MTT assay, wound scratch assay and Transwell assay, apoptosis assay in vitro and xenograft model in vivo. mRNA-seq and CHIP-seq was used to explore the molecular mechanism of biological function of EZH2. mRNA and H3K27me3 profiles of wild type (WT) SKOV3 and SKOV3/sgEZH2 were generated by deep sequencing

卵巢癌是妇科恶性肿瘤中死亡率最高的癌种，且多数患者确诊时已处于进展期。EZH2一直是卵巢癌重要的肿瘤标志物与有效的治疗靶点，但其分子机制尚未明确。本研究通过体外与体内实验，探究EZH2敲除的SKOV3细胞的生物学效应，并结合mRNA测序（mRNA-seq）与染色质免疫沉淀测序（CHIP-seq）的整合分析，解析其背后的分子机制。
本研究采用CRISPR/Cas9系统构建EZH2敲除的SKOV3细胞系。通过蛋白质印迹（Western blot）检测相关蛋白的表达水平。分别利用MTT法、划痕愈合实验、Transwell实验及细胞凋亡实验，检测体外环境下EZH2对卵巢癌细胞的生物学功能影响，并通过体内异种移植瘤模型验证上述效应。采用mRNA-seq与CHIP-seq联合分析，探究EZH2调控卵巢癌生物学功能的分子机制。通过深度测序获取野生型（WT）SKOV3细胞与SKOV3/sgEZH2细胞的mRNA及H3K27me3修饰图谱。