缺氧影响胃腺癌循环肿瘤细胞发生的体内实验  

Effects of hypoxia on circulating tumor cells in gastric adenocarcinoma in vivo

作　　者：李熳[1] 张志广[1] 刘霞[1] 张雪莲[1] 

机构地区：[1]天津医科大学第二医院消化内科,300211

出　　处：《中华消化杂志》2014年第3期156-159,共4页Chinese Journal of Digestion

基　　金：天津市高等学校科技发展基金计划（20110130）;天津医科大学科技基金（2008ky28）

摘　　要：目的探讨微环境缺氧的变化与胃腺癌循环肿瘤细胞（CTC）发生之间的相关性。方法将人胃腺癌BGC-823细胞用不同终浓度CoCl2（0、100、200、300、400μmol／L）处理4h，应用Western印迹法检测缺氧诱导因子-la（HIF-la）表达水平来筛选最适浓度；将40只BALB／c小鼠平均分为缺氧组与常氧组，分别采用尾静脉注射经上述最适浓度处理的BGc～823细胞与未经缺氧诱导的BGC-823细胞，3d后取内呲血，采用密度梯度离心法富集CTC，以细胞角蛋白19（CKl9）抗体为标志物行细胞免疫荧光鉴定并计数CTC。采用方差分析和Dunnett-T法进行组间差异分析。结果BGC-823细胞经终浓度200μmol／LCoCI2模拟缺氧4h后HIF-1a表达水平（1．40土0．16）最高。缺氧组小鼠CTC检出数量较常氧组增高[（4．70±1．72）／N倍视野比（1．15±0．56）／高倍视野，z=-4．824，P〈0．01]。结论微环境缺氧可能促进胃腺癌出现CTC。Objective To explore the correlation between the change of hypoxic in microenvironment and the genesis of circulating tumor cells （CTCs） in gastric adenocarcinoma. Methods After human gastric adenocarcinomn BGC-8Z3 cells treated with different concentration of CoCl2 （0,100, 200,300,400 /1tool/L） for four hours, the expression levels of hypoxia inducible factor 1 alpha （HIF-1α） were detected by Western blot to select the optimal concentration. A total of 40 BALB/c mice were divided into two equal groups, named hypoxic group and normoxie group. Optimal concentration of COC12 treated BGC-823 gastric cancer cells and non-hypoxic induced BGC-823 gastric cancer cells were injected throughthe tail vein. After three days blood samples were collected through the angular vein. CTCs were enriched by density gradient eentrifugation, identified with cytokeratin19 （CK19） antibody marker by cell immunofluorescence and cell number was counted. Statistical significace test were performed by analysis of variance and Dunnett-T test. Results The expression of HIF-I~ reached a maximun of 1.40±0.16 when BGC-823 cells were treated with 200 tlmol/L COC12 for four hours. The number of CTCs in hypoxic group was higher than that of normoxic group （（4.70±1.72）/high power field （HPF） vs （1.15±0.56）/HPF, Z=-4. 824,P〈0.01）. Concision Hypoxic microenvironment might promote the genesis of CTC in gastric adenocarcinoma.

关 键 词：缺氧诱导因子1 胃肿瘤 腺癌 循环肿瘤细胞 肿瘤转移 

分 类 号：R735.2[医药卫生—肿瘤]