Id2通过非HLH结构域促进MCF-7细胞的侵袭性  

A HLH domain deletion of Id2 promotes migration in MCF-7 breast cancer cells

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室,北京100853

出　　处：《现代肿瘤医学》2008年第12期2043-2046,共4页Journal of Modern Oncology

基　　金：国家自然科学基金项目(编号:30670809)

摘　　要：目的:观察Id2基因转染乳腺癌细胞MCF-7后对细胞生长、侵袭能力的变化,探讨Id2基因缺失HLH结构域后对细胞的影响。方法:以携带Id2-DBM和Id2-DBM-δHLH基因的质粒(pCDNA3.1-Id2-DBM和pCDNA3.1-Id2-DBM-δHLH)经脂质体介导转染MCF-7细胞。RT-PCR法检测转染后Id2、Id2-DBM和Id2-DBM-δHLH mRNA表达水平;MTT法检测MCF-7细胞增殖曲线;划痕实验、transwell小室检测细胞的迁移能力;Western Blot分析Id2对MCF-7细胞E-cad表达的影响。结果:mRNA水平显示质粒均成功转入;细胞生长曲线显示增殖无显著差异;与空载组比较,转染pCDNA3.1-Id2-DBM和pCD-NA3.1-Id2-DBM-δHLH后侵袭能力增强,且E-cad表达降低。结论:Id2蛋白的过表达可以促进乳腺癌细胞MCF-7的侵袭能力,与E-cad的降低有关,且该作用在缺失HLH结构域后仍然存在。Objective ：To study the influence of Id2 on cell proliferation and migration in MCF -7 cells, and to investigate the influence of HLH - deleted Id2. Methods： Human breast cancer cell line MCF - 7 was transfected with peDNA3.1 - Id2 - DBM and pcDNA3.1 - Id2 - DBM - 8HLH vectors by SuperFect Transfeetion Reagent, and cells transfected with blank vector pCDNA3.1 were used as control, mRNA of ld2, Id2 - DBM and Id2 - DBM - ~HLH were measured by reverse transcription - polymerase chain reaction （ RT - PCR）. Cell proliferation was as- sessed by MTr assay. Cell invasion was detected by scratch assay and transwell chaber assay. The regulation of pro- tein E -cadherin was observed by Western blot after transfection. Results： The cellular proliferation had no novel changes before and after transfection （ P 〉 0.05 ）. We found that overexpression Id2 - DBM and Id2 - DBM - 8HLH promoted MCF - 7 cells migration into the wound by scratch assay. Furthermore, the number of migrated cells was in- creased by transwell chamber assay （P 〈 0. 001 ）. In addition, the E - eadherin protein was downregulated after transfeetion compared to control. Conclusion： Overexpression of protein Id2 promoted migration in MCF -7 cells. This effect might be related with the decrease of E - cadherin, and it still retained after the HLH domain of Id2 was deleted.

关 键 词：分化抑制因子蛋白2 螺旋-环-螺旋构型 乳腺癌 细胞侵袭 上皮钙黏附素 

分 类 号：R730[医药卫生—肿瘤]