CD133^＋肺癌干细胞中基质金属蛋白酶9和缺氧诱导因子2α的表达及其病理学机制  

Expression and pathological mechanism of MMP-9 and HIF-2α in CD133^＋ lung cancer stem cells

作　　者：高远[1] 冯军[1] 吴灵芝[1] 詹升华[2] 孙静[3] 

机构地区：[1]苏州大学附属第一医院肿瘤科,215000 [2]苏州大学附属第一医院病理科,215000 [3]苏州大学基础医学与生物科学学院免疫学系

出　　处：《中华医学杂志》2015年第32期2607-2611,共5页National Medical Journal of China

基　　金：国家自然科学基金（31100634）;苏州市科技计划项目（SYS201464）

摘　　要：目的 探讨CD133^＋肺癌干细胞中基质金属蛋白酶（MMP）-9和缺氧诱导因子（HIF）-2α的表达及其病理学机制.方法 选取2009年1至12月苏州大学附属第一人民医院收治的肺癌手术患者组织石蜡标本62例.运用免疫组织化学染色技术检测CD133分子在肺癌组织中的表达,分析其临床意义;实时荧光定量PCR法分析CD133^＋肺癌干细胞中MMP-1、MMP-2、MMP-9、HIF-1α、HIF-1β、HIF-2α和MMP组织抑制剂（TIMP）-1、TIMP-2、TIMP-3、TIMP-4等转移相关基因的表达;分别用乱序siRNA和CD133 siRNA转染肺癌细胞株A549,构建对照-si-A549细胞和CD133-si-A549细胞,PCR法分析两组细胞中CD133、MMP-9和HIF-2α基因的表达;肿瘤侵袭试验分析细胞侵袭迁移能力的改变.结果 51.6％的肺癌组织阳性表达CD133分子,其表达与肺癌的转移和患者生存率密切相关（P＜0.05）.CD133^＋与CD133-细胞中TIMP-1、TIMP-2、TIMP-3、TIMP-4、HIF-1α、HIF-1β、MMP-1、MMP-2相对表达量差异均无统计学意义（均P＞0.05）;CD133^＋细胞中HIF-2α和MMP-9的相对表达量均显著高于CD133-细胞（1.58±0.39比1.10±0.31,1.67±0.38比1.05±0.21,均P＜0.05）.CD 133-si-A549细胞中CD133、HIF-2α和MMP-9相对表达量均显著低于对照-si-A549细胞（0.24±0.10比0.85±0.23、0.19±0.09比0.54±0.18和0.31±0.17比1.12±0.31,均P＜0.05）.CD133-si-A549细胞迁移至下室细胞数量显著低于对照-si-A549细胞（207±25比82±10,P＜0.05）.结论 CD133^＋肺癌干细胞与肿瘤转移和患者预后相关,它可通过上调HIF-2α和MMP-9的表达促进肿瘤的转移.Objective To investigate the expression and pathological mechanism of matrix metalloproteinase （MMP）-9 and hypoxia-inducible factor （HIF）-2α in CD133^＋ lung cancer stem cells.Methods Sixty-two cases of lung cancer paraffin embedding tissues were collected from the First Affiliated Hospital of Soochow University between January 2009 and December 2009.Immunohistochemistry （IHC） was used for detection of CD133 expression in lung cancer tissues and the clinical significance was analyzed.Real-time polymerase chain reaction （PCR） was used for the investigation of expression of tumor metastasis associated genes,including MMP-1,MMP-2,MMP-9,HIF-1α,HIF-1β,HIF-2α and tissue inhibitor of metalloproteinase （TIMP）-1,TIMP-2,TIMP-3,TIMP-4.Scrambled siRNA or CD133 siRNA were used to transfect the lung cancer cell line A549,then Control-si-A549 cells and CD133-si-A549 cells were generated respectively.PCR was used to analysis CD133,MMP-9 and HIF-2α genes expression and transwell invasion assay was used to study the invasion ability of A549 cells in two groups.Results 51.6% of lung cancer tissues expressed CD133 （P 〈 0.05）;the expression level of CD133 was related to tumor metastasis and patients＇ survival rate （P 〈0.05）.The gene expression of HIF-2α and MMP-9 were increased in CD133^＋ lung cancer cells compared with CD133-cancer cells （1.58 ± 0.39 vs 1.10 ± 0.31,1.67 ± 0.38 vs 1.05 ±0.21,all P 〈0.05）,whereas no difference was found in gene expression of MMP-1,MMP-2,HIF-1 α,HIF-1β and TIMP-1,TIMP-2,TIMP-3,TIMP-4 （all P 〉 0.05）.Compared with the Control-si-A549 cell,the expression of CD133,HIF-2α and MMP-9 （0.24 ±0.10 vs 0.85 ±0.23,0.19 ±0.09 vs 0.54 ± 0.18,0.31 ± 0.17 vs 1.12 ± 0.31,all P 〈 0.05） in CD133-si-A549 cell were remarkably decreased.The number of CD133-si-A549 cells migrated to below room was significantly smaller than that of Control-si-A549 cells （207 ± 25 vs 82 ± 10,P 〈 0.05）.Concluslons The CD133 ＋ lung cancer stem cell is correlated

关 键 词：肺肿瘤 肿瘤干细胞 CD133 基质金属蛋白酶9 缺氧诱导因子2 α亚基 

分 类 号：R734.2[医药卫生—肿瘤]