ALDH1A2基因过表达对结肠癌LoVo细胞增殖、侵袭及黏附能力的影响  

Effect of overexpresion of ALDH1A2 gene on LoVo cell invasion and metastasis

作　　者：杨万广[1] 张谢夫[2] 张善锋[3] 

机构地区：[1]郑州大学第一附属医院急诊外科,450052 [2]郑州大学第一附属医院胃肠外科 [3]郑州大学基础医学院生物化学与分子生物学教研室

出　　处：《中华实验外科杂志》2012年第9期1747-1749,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察ALDH1A2基因过表达对LoVo细胞增殖、侵袭及黏附能力的影响。方法将携带有人外源性ALDH1A2基凶的真核表达质粒转染人结肠癌细胞株LoVo细胞，分别应用逆转录-聚合酶链反应（RT—PCR）和Westernblot法从mRNA及蛋白水平测定3组细胞中ALDH1A2水平变化，通过噻唑蓝（MTF）、Matrige!体外侵袭实验、平板克隆形成实验、黏附实验测定不同组别细胞增殖能力、侵袭转移能力、克隆形成能力及黏附能力的变化。结果RT—PCR及Westernblot结果显示，与空质粒转染组及空白对照组细胞比较，转染pEGFP—NI—ALDH1A2的细胞其内源性ALDHlA2mRNA及蛋白表达水平显著增加（P〈0．05）；MTY及平板克隆实验显示转染pEGFP-N1-ALDH1A2质粒的LoVo增殖能力显著下降；Matrigel体外侵袭实验证实，转染pEGFP—N1-ALDH1A2质粒的LoVo穿膜细胞数为（21．4±3．1）个，显著低于转染空质粒及空白组，LoVo穿膜细胞数为（71．9±7．27、78．3±9．35）个，差异有统计学意义（P〈0．05），黏附实验结果显示，转染pEGFP—N1-ALDHIA2质粒的LoVo黏附能力显著降低（P〈0．05），提示转染pSUPER—TF-1质粒的LoVo细胞侵袭、黏附能力显著下降。结论ALDH1A2基因过表达可以明显降低LoVo细胞增殖能力、体外侵袭黏附能力。Objective To evaluate the effects of ALDHIA2 gene over-expression on human LoVo cell line proliferation, invadesion and adhesion. Methods Human eukaryotie expression plasmid pEGFP- N1-ALDH1A2 were transfeeted to LoVo cell line. Reverse transeription-polymerase chain reaction （RT- PCR） and Western blotting were used to detect the changes of ALDH1 A2 gene expression,A methyl thiazol tetrazolium （MTT） assay, Matrigel invasion assay, plate clony-forming test and adhesion test were performed to assess the LoVo cells＇ proliferative activity, invadsion ability, elony-forming capability and adhesive abili- ty before and after transfeetion. Results RT-PCR and Western blotting analysis showed that LoVo cells with pEGFP-N1-ALDH1A2 transfected increased the ALDH1A2 gene expression compare（] with control cells. ALDH1 A2 over-expression can reduce LoVo cells proliferation,invadsion ability,clony-forming capability and adhesive ability （P 〈 0. 05 ）. Conclusion ALDH1 A2 over-expression can inhibit the proliferation and elony-forming ability of human LoVo cell.

关 键 词：结肠癌 ALDH1A2 克隆 侵袭 黏附 

分 类 号：R735.35[医药卫生—肿瘤]