抑瘤基因NGX6对人结肠癌细胞HT-29生长的影响  

Effects of tumor suppressor gene NGX6 on growth of human colon cancer cell line HT-29

作　　者：王晓艳[1] 沈守荣[1] 刘华英[2] 张晓梅[3] 彭聪[2] 黄河[2] 刘芬[1] 李晓玲[2] 李桂源[2] 

机构地区：[1]中南大学湘雅三院消化内科,湖南省长沙市410013 [2]中南大学湘雅医学院肿瘤研究所,湖南省长沙市410078 [3]中南大学湘雅医院消化内科,湖南省长沙市410078

出　　处：《世界华人消化杂志》2004年第3期574-579,共6页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目.No.30370801湖南省自然科学基金资助项目;No.01JJY2102和03JJY2004~~

摘　　要：目的:NGX6是新克隆的侯选抑瘤基因,其在结肠癌组织中表达明显下调,提示NGX6的差异表达与结肠癌发生有关.本文通过研究NGX6对人结肠癌细胞HT-29生物学特性的影响以明确NGX6在结肠癌发生发展中的作用. 方法:脂质体介导pcDNA3.1(+)/NGX6重组体转染低表达NGX6的人结肠癌细胞HT-29,Dot blot及RT-PCR方法检测外源性NGX6基因的表达,借助生长曲线、MTT、软琼脂集落形成、流式细胞仪、裸鼠成瘤实验对NGX6转染前后人结肠癌细胞HT-29的生物学行为进行检测. 结果:pcDNA3.1(+)/NGX6/HT-29组细胞生长速度比pcDNA3.1(+)/HT-29和HT-29组明显减慢(P<0.05),其倍增时间为34.5 h比pcDNA3.1(+)/HT-29(27.1 h)和HT-29 (23.0 h)延长;pcDNA3.1(+)/NGX6/HT-29组软琼脂集落形成率较对照组明显下降(P<0.05);流式细胞仪检测NGX6高表达能延缓HT-29细胞周期由G0/G1期→s期的进程;裸鼠成瘤实验表明pcDNA3.1(+)/NGX6/HT-29组成瘤受抑. 结论:NGX6的重表达可逆转人结肠癌细胞HT-29的恶性表型;NGX6是极具前途的结肠癌相关抑瘤基因侯选者.AIM: NGX6 is a novel tumor suppressor gene candidate, and the expression of NGX6 decreased in colon cancer specimen, which suggested that the down-regulation of NGX6 was closely related to tumorigenesis of colon cancer. This study was designed to explore effects of NGX6 on the cytobiological characteristics of human colon cancer cell line HT-29 and to elucidate the role of NGX6 in the colon cancer. METHODS: The mammal expression vector of pcDNA3.1 (+)/NGX6 was transfected into human colon cancer cell line HT-29 cells by liposome.The integration of the exogenous vector DNA and the expression of NGX6 were detected by dot blot and RT-PCR. Finally, the cytobiological characteristics of positive clone were analyzed by growth curves of cells, MTT, soft agar assay, FCM and xenograft. RESULTS: pcDNA3.1(+)/NGX6/HT-29 cells grew slower than pcDNA3.1(+)/HT-29 and HT-29 cells (P<0.05), and the PDT of pcDNA3.1(+)/NGX6/HT-29 cell was 34.5 hours, which was significantly longer than that of pcDNA3.1(+)/ HT-29 cells (27.1 h) and HT-29 cells (23.0 h). FCM data showed that the overexpression of NGX6 blocked the progression of G1→S phase in cell cycle, and it also presented decreased clonogenicity in soft agar assay and tumor formation in nude mice. CONCLUSION: The re-expression of NGX6 can reverse the malignant phenotype of human colon cancer cell line HT-29. Therefore, NGX6 gene may be a good candidate of tumor suppressor gene associated with colon cancer.

关 键 词：NGX6基因 肿瘤抑制 结肠癌 HT-29细胞 肿瘤生物学 DNA探针 

分 类 号：R735.35[医药卫生—肿瘤]