Id2通过非HLH结构域促进SKOV3细胞的侵袭性  

Id2 promotes migration of SKOV3 ovarian cancer cells by non-HLH domain

作　　者：顾成磊[1] 伍志强[2] 孟元光[1] 赵亚力[2] 杨洁[2] 韩为东[2] 

机构地区：[1]解放军总医院妇产科,北京100853 [2]解放军总医院分子生物学室,北京100853

出　　处：《肿瘤》2008年第9期767-770,共4页Tumor

基　　金：国家自然科学基金资助项目(编号:30670809)

摘　　要：目的:观察分化抑制因子2(inhibitor of differentiation 2,Id2)基因转染卵巢癌SKOV3细胞后,对细胞生长和侵袭能力的影响,探讨Id2基因缺失螺旋-环-螺旋(helix-loop-helix,HLH)结构域后对SKOV3细胞的影响。方法:以携带野生型Id2、Id2-DBM和Id2-DBM-δHLH基因的质粒(pcDNA3.1-Id2、pcDNA3.1-Id2-DBM和pcDNA3.1-Id2-DBM-δHLH)经脂质体介导转染SKOV3细胞。采用Western印迹法和RT-PCR法检测转染后SKOV3细胞Id2、Id2-DBM和Id2-DBM-δHLH的表达水平;MTT法检测SKOV3细胞的增殖曲线;划痕试验和Transwell小室检测细胞的迁移能力;Western印迹法检测Id2对MCF-7细胞上皮钙黏附素表达的影响。结果:mRNA及蛋白水平显示质粒成功转入;细胞生长曲线显示各组细胞的增殖无显著差异;与空白对照组相比,转染pcDNA3.1-Id2、pcDNA3.1-Id2-DBM和pcDNA3.1-Id2-DBM-δHLH后,细胞的侵袭能力增强,且pcDNA3.1-Id2-DBM和pcDNA3.1-Id2-DBM-δHLH组细胞明显伴有上皮钙黏附素表达水平的降低。结论:Id2蛋白的过表达可促进卵巢癌SKOV3细胞的侵袭能力,与上皮钙黏附素表达水平的降低有关,且该作用在缺失HLH结构域后依然存在。Objective ： To study the influence of inhibitor of differentiation 2 （ Id2 ） on cell proliferation and migration in SKOV3 cells, and to investigate the influence of HLH domain deletion in Id2 gene on SKOV3 cells. Methods： Human ovarian cancer cell line SKOV3 was transfeeted with pcDNA3.1-Id2, pcDNA3.1-Id2-DBM and peDNA3.1-Id2-DBM-SHLH vectors by SuperFeet Transfeetion Reagent. The cells transfected with blank vector pCDNA3. 1 were used as control. Id2 protein was detected by Western blot, and mRNA transcriptions of Id2, Id2-DBM （ D-box mutant） , and Id2-DBM-δHLH were measured by RT-PCR. Cell proliferation was assessed by MTT assay. Cell invasion was detected by scratch test and transwell chamber assay. The regulatory effect of Id2 on E-cadherin was evaluated by Western blot. Results： The mRNA and protein levels indicated that the plasmid was successfully transfected. Cell growth curve suggested that the proliferation of cells had no significant difference between each group. Compared with the control group, cell migration ability was significantly enhanced after transfection with pcDNA3.1-Id2, pcDNA3.1-Id2-DBM and pcDNA3. 1-1d2-DBM- δHLH vectors. The downregulation of E-cadherin protein was accompanied with increased migration capability in pcDNA3.1-1d2-DBM and pcDNA3.1-1d2-DBM-δHLH vectors-transfeeted cells. Conclusion： Overexpression of Id2 promotes the migration capability of SKOV3 cells, which might be related with the downregulation of E-cadherin. The action still exists after the HLH domain deletion in Id2 gene.

关 键 词：分化抑制因子 卵巢肿瘤 肿瘤侵袭 钙黏着糖蛋白类 

分 类 号：R73-377[医药卫生—肿瘤] R737.31[医药卫生—临床医学]