CST1靶向OTUB1对胃癌干细胞特性的影响  

Effect of CST1 on cancer stem cell-like properties in gastric cancer cells via targeting OTUB1

作　　者：陈雪霁[1] 丁玉霞 朱波 CHEN Xue-ji;DING Yu-xia;ZHU Bo(Department of Oncology,North China Petroleum Hospital,Cangzhou 062552,China)

机构地区：[1]华北石油管理局总医院肿瘤科,河北沧州062552

出　　处：《解剖科学进展》2024年第6期641-644,共4页Progress of Anatomical Sciences

基　　金：河北省2021年度医学科学研究项目(20210591)。

摘　　要：目的探讨半胱氨酸蛋白酶抑制剂1(CST1)能否通过靶向去泛素化酶OTUB1影响胃癌干细胞样特性。方法RT-PCR检测CST1、OTUB1在34例胃癌组织及癌旁组织中的表达,双荧光素酶实验验证CST1与OTUB1靶向结合作用,选择人胃癌细胞株MKN-45采用免疫磁珠法从中分选出干细胞,并随机分为si-CST1组、si-NC组和Blank组(空白对照),肿瘤球形成实验、克隆形成实验检测细胞干性,CCK8检测细胞增殖能力,Western blot检测细胞CST1、OTUB1及细胞干性标志物CD133、CD44、Nanog、Bmi1蛋白表达。结果胃癌组织中CST1、OTUB1高表达,CST1与OTUB13′-UTR存在碱基互补区域;下调CST1表达后,细胞OTUB1蛋白表达、胃癌干细胞成球数、克隆形成数目、增殖水平、干性标志物CD133、CD44、Nanog、Bmi1蛋白表达均降低。结论下调CST1表达可抑制胃癌干细胞自我更新和增殖能力,是通过靶向调控OTUB1表达起作用的。Objective To investigate whether cysteine protease inhibitor 1(CST1)can affect stem cell-like properties of gastric cancer by targeting deubiquitinase OTU domain-containing ubiquitin aldehyde-binding protein 1(OTUB1).Methods The expressions of CST1 and OTUB1 in 34 gastric cancer tissues and adjacent tissues was detected by RT-PCR.The targeted binding effect of CST1 and OTUB1 was verified by double luciferase assay.The human gastric cancer cell line MKN-45 was selected.The stem cells were purified by the immunomagnetic bead method,and classified into si-CST1 group,si-NC group and Blank group(blank control).The sphere-forming capacity assay and colony formation assay were performed to evaluate the cell stemness.CCK8 was conducted to test cell proliferation,and Western blot was used to measure the expressions of CST1,OTUB1,CD133,CD44,Nanog and Bmi1 in cells.Results The expression of CST1 and OTUB1 were up-regulated in gastric cancer tissues,and there was a base complementary between CST1 and OTUB13′-UTR regions.After the down-regulation of CST1,the expression of OTUB1,the number of ball formation,the number of clone formation,the proliferation level,and the expressions of dry markers CD133,CD44,Nanog and Bmi1 were all decreased.Conclusion Down-regulation of CST1 expression can inhibit self-renewal and proliferation of gastric cancer stem cells,which may be related to the its targeted regulation of OTUB1 expression.

关 键 词：CST1 去泛素化酶 胃癌 干细胞样特性 

分 类 号：R735.2[医药卫生—肿瘤]