论文部分内容阅读
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 investigate the effect of E-cadherin and intercellular adhesion molecule-1 (ICAM-1) on multidrug resistance of colon cancer HT-29 cells and its possible mechanism. Methods: Three-dimensional or monolayer cultures of HT-29 cells were treated with liq-uid overlay technique or conventional attachment method, respectively. The radial out-growth assay or MTT assay was used to determine drug sensitivity. Immunohistochemistry and Western blotting were used to detect the expression of E-cadherin, The expression of ICAM-1 protein was detected by immunohistochemistry. Apoptosis was detected by TUNEL. The expression of apoptosis-related proteins (Caspase-3, Bcl-2 and Bax) was detected by Western blotting. Results: Compared with monolayer cultured HT-29 cells, the sensitivity of 5-FU to HT-29 cells was decreased (18.74 ± 0.97) and (1.38 ± 0.09) μg / mL respectively. The expressions of E-cadherin and ICAM- 1 expression increased. After anti-adhesion treatment with different concentrations of Hyaluronidase (Hyase), HT-29 cells in three-dimensional culture increased the sensitivity to 5-FU (the IC50 value was significantly decreased), the expression of E-cadherin and ICAM- The number increased, Caspase-3 activation fragment and Bax expression increased, while Bcl-2 expression decreased, and showed a concentration-dependent characteristics. Conclusion: E-cadherin and ICAM-1 can induce multidrug resistance in HT-29 colon cancer cells, which may be related to the regulation of apoptosis-related proteins.