体外三维培养卵巢癌A2780细胞及其多药耐药机制的实验研究  

Study on Three-dimensional Culture A2780 Cell of Ovarian Carcinoma and Drug Resistance Mechanism

作　　者：高爱莲 

机构地区：[1]尉氏县计划生育指导站,河南尉氏4750011

出　　处：《河南职工医学院学报》2008年第3期222-225,共4页Journal of Henan Medical College For Staff and Workers

摘　　要：目的建立卵巢癌A2 780细胞形成的多细胞团簇模型,探讨凋亡在卵巢癌细胞群集耐药中的作用。方法以三维培养方法获得人卵巢癌A 2 780多细胞球体(MCS)模型,单层细胞(MC)为对照;MTT法和liquid over-lay技术检测细胞的药物敏感性;流式细胞术检测细胞周期和凋亡的变化;RT-PCR分析凋亡相关基因caspase-3和bcl-2的表达变化。结果细胞在培养2 d后即可形成由多个细胞组成的细胞团,培养5 d后,形成有多层细胞结构的细胞球;与A2 780/MC相比,A 2 780/MCS细胞药物敏感性及细胞凋亡率均显著下降,差异有显著性;而与A2 780/MC相比,凋亡诱导基因caspase-3在A2 780/MCS细胞中明显下降,凋亡抑制基因bcl-2明显增高(P<0.05)。结论卵巢癌细胞群集耐药可能与肿瘤细胞凋亡相关基因caspase-3和bcl-2的表达相关。Objective To explore an optimal model of multicellular aggregates of ovarian cancer cell line A2780 and investigate the possible mechanism of ovary cancer multicellular drug resistance. Methods Three-dimensional culture was used to form MCS of human ovarian cancer cell line A2780. Monolayer A2780 cell was cultured as control. Drug sensitivity of monolayer cells （MC） and multicellular spheroids （MCS） were tested by MTT staining and radial outgrowth assay respectively. The apoptosis efficiency was determined by flow cytometry （FCM）. The expression of caspase-3 and bcl-2 in A2780/ MC and A2780/MCS were detected by RT-PCR. Results A2780/MC was compacted into mass after 2 days in three-dimensional cell culture model, and MCS had more than two layers of cells growing within 5 days. Compared with A2780/MC, A2780/MCS were more resistant to the anticancer drug, and the apoptosis rate was significantly lower than those of A2780/MC. The expression of caspase-3 in A2780/ MCS cells was sharply higher than that of A2780/Mc cells, but the expression of bcl-2 was significantly lower than that of A2780/Mc cells （P 〈 0.05）. Conclusion The drug resistance of MCS may be associated with the expression of apoptosis correlated gene caspase-3 and bcl-2.

关 键 词：多细胞团簇 卵巢肿瘤 CASPASE-3 BCL-2 

分 类 号：R969.4[医药卫生—药理学]