RNA结合基序5基因敲除子宫内膜癌细胞系及其作用研究  

RBM5 knockout in endometrial cancer cells and its effect

作　　者：王赛凤 史蔚 谢咸晶 WANG Saifeng;SHI Wei;XIE Xianjing(The International Peace Maternity and Child Health Hospital,School of Medicine,Shanghai Jiao Tong University,Shang-hai Key Laboratory of Embryo Original Diseases,Shanghai 200030,China)

机构地区：[1]上海交通大学医学院附属国际和平妇幼保健院,上海市胚胎源性疾病重点实验室妇科,上海200030

出　　处：《实用医学杂志》2022年第10期1203-1207,共5页The Journal of Practical Medicine

基　　金：上海市卫生健康委员会资助项目(编号:20194Y0050)。

摘　　要：目的探讨用CRISPR/Cas9系统敲除RNA结合基序5(RBM5)基因对子宫内膜癌细胞生物学行为的影响。方法构建CRISPR/Cas9⁃RBM5⁃sgRNA重组质粒并转染于高表达RBM5的Ishikawa细胞系后,Western Blot、qRT⁃PCR分析RBM5表达并进行Sanger测序。CCK⁃8、克隆形成和Transwell检测其对子宫内膜癌细胞增殖、迁移和侵袭能力的影响。结果敲除细胞系中RBM5表达降低(P<0.05);测序结果表明在RBM5编辑位点产生移码突变。RBM5敲除细胞株增殖、迁移和侵袭能力显著增强(P<0.05)。结论RBM5缺失促进子宫内膜癌细胞增殖、迁移和侵袭,本研究为子宫内膜癌分子调控机制的研究提供了参考依据。Objective To investigate the effect of RBM5 gene knockout by CRISPR/Cas9 system on the biological behavior of endometrial cancer cells.Methods CRISPR/Cas9⁃RBM5⁃sgRNA recombinant plasmids were transfected intoRBM5⁃high expression Ishikawacell lines.Western blot,qRT⁃PCR and Sanger sequencing were used to verify the RBM5 knockout.The abilities of proliferation,migration and invasion of parental strains,knock⁃out strains and revertant strains were analyzed by CCK⁃8 kit,colony formation and Transwell assays.Results The expression of RBM5 was significantly decreased in the knockout cell lines(P<0.05).The sequencing results showed a code shift mutation.The abilities of proliferation,migration and invasion in the RBM5⁃knockout cell significantly increased(P<0.05).Conclusion RBM5 knockout in endometrial cancer cell lines can promote cell proliferation,migration and invasion,which guides further molecular mechanism of endometrial cancer.

关 键 词：CRISPR/Cas9 子宫内膜癌 RBM5 增殖 迁移 侵袭 

分 类 号：R73-34[医药卫生—肿瘤]