胃癌细胞株MKN45球体细胞中Sox2的表达研究

摘要

Objective:To detect the expression of stem-cell related factor Sox2 in spheroid body-forming cells of gastric cancer cell line MKN45. Methods: We used gastric cancer cell line MKN45 to culture spheroid bodies in nonadherent condition in a serum-free medium supplemented with epidermal growth factor(EGF) and basic fibroblast growth factor(bFGF),and evaluated their resistance to DDP. Using reverse transcriptase polymerase chain reaction(RT-PCR) and Western Blot analysed the expres-sion level of the embryonic stem cell-related gene Sox2 in spheroid body forming cells and parental cells. Results: In this study, we observed that MKN45 cells formed spheroid bodies in nonadherent condition in a serum-free medium. The spheroid body-forming cells possessed the ability of chemoresistance, and showed higher expression of the embryonic stem cell-related gene Sox2. Conclusion: Spheroid body-forming cells developed from human gastric cancer cell line MKN45 in nonadherent condition in a serum-free medium supplemented with EGF and bFGF acquired CSCs characteristics. Spheroid body formation assay may be a practical and effective approach to isolate and identify CSCs.%目的：检测干细胞相关因子Sox2在胃癌细胞株MKN45悬浮球体细胞中的表达。方法：选择人胃癌细胞株MKN45，在含有表皮生长因子(epidermal growth factor，EGF)及基本的成纤维细胞生长因子(basic fibroblast growth factor，bFGF)无血清培养液中培养，观察球体形成情况，并检测球体细胞对化疗药物顺铂(cisplatin，DDP)的耐受性；采用RT-PCR与免疫印迹分析检测干细胞相关基因Sox2在球体细胞及原代贴壁细胞两组细胞中的表达差异。结果：胃癌细胞株MKN45在无血清培养条件下能形成悬浮球体，球体细胞中Sox2的相对表达量明显高于原代贴壁细胞(P=0.000)。结论：悬浮球培养法在胃癌细胞株MKN45中培养形成的球体细胞具有肿瘤干细胞特性。该方法是分离和鉴定肿瘤干细胞的实用而有效的方法。