E-钙黏素和ICAM-1诱导结肠癌多细胞耐药的实验研究  

Multicellular resistance to 5-FU induced by E-cadherin and ICAM-1 in colon carcinoma cells

作　　者：李建军[1] 潘凤[1] 胡绍毅[1] 黄海辉[1] 边志衡[1] 梁后杰[1] 

机构地区：[1]第三军医大学西南医院肿瘤科,重庆400038

出　　处：《中华肿瘤防治杂志》2009年第2期85-88,120,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30171067);重庆市科技攻关计划项目(9398)

摘　　要：目的:探讨E-钙黏素(E-cadherin)和细胞间黏附分子-1(intercellular adhesionmolecule-1,ICAM-1)诱导结肠癌HT-29细胞多细胞耐药的作用及可能的机制。方法:HT-29细胞的三维或单层培养分别采用liq-uid overlay技术或常规贴壁法;radial out-growth分析或MTT法测定药物敏感性;免疫组织化学和蛋白质印迹法检测E-cadherin、ICAM-1蛋白的表达;TUNEL检测细胞凋亡;蛋白质印迹法检测凋亡相关蛋白(Caspase-3、Bcl-2和Bax)的表达水平。结果:三维培养HT-29细胞与单层培养细胞相比,对5-FU的敏感性降低,IC50值分别为(18.74±0.97)和(1.38±0.09)μg/mL;E-cadherin、ICAM-1表达增强。以不同浓度透明质酸酶(Hyaluronidase,Hyase)抗黏附处理后,三维培养HT-29细胞对5-FU敏感性增加(IC50值明显降低),E-cadherin和ICAM-1表达减弱,凋亡细胞数目增多,Caspase-3活化片断和Bax表达增强,而Bcl-2表达减少,并呈现浓度依赖性的特点。结论:E-cadherin和ICAM-1可诱导结肠癌HT-29细胞产生多细胞耐药,其机制可能与凋亡相关蛋白的表达调控有关。OBJECTIVE： To explore the effect of E-cadherin and intercellular adhesion molecule-1 （ICAM-1） on muhicellular resistance （MCR） to 5 FU in colon carcinoma ceils and its mech anisms. METHODS： The muhicellular spheroids （MCS） of colon carcinoma HT-29 cell line were cultured by the liquid overlay technique and monolayer cells by using the routine procedure. The 50% inhibiting concentration （IC50） of spheroids or monolayer cells for 5-FU was determined by the radial outgrowth assay or MTT assay respectively and the index of MCR （MCRI） was calculated. The expressions of E-cadherin and ICAM-1 in spheroids or monolayer cells were detected by immunohistochemistry and Western blotting. Apoptosis was examined by TUNEL. Caspase-3, Bcl 2 and Bax were detected by Western blotting. RESULTS： The value of IC50 for 5-FU was （ 18.74± 0.97） μg/mL in MCS while （1.38＋0.09） μg/mL in monolayer cells. The expressions of E-cadherin and ICAM-1 in MCS were also up-regulated. After HT-29 MCS were treated by hyaluronidase with different concentrations, the value of IC50 for 5-FU was decreased and the expression of E-cadherin or ICAM-1 was reduced. Meanwhile, the apoptotic rate was increased, the expression of Caspase-3 large subunit and Bax were up regulated and Bcl 2 was decreased. CONCLUSION： E-cadherin and ICAM-1 can induce MCR to 5 FU in colon carcinoma HT 29 cells which might involved in the mediation of apoptotic proteins.

关 键 词：结肠肿瘤 E-钙黏素 细胞间黏附分子-1 多细胞耐药 细胞凋亡 

分 类 号：R735.35[医药卫生—肿瘤]