CAJ | 학술논문

目的：检测干细胞相关因子CD44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,在含有纤维生长因子-2及表皮生长因子无血清培养液中培养，观察球体形成情况；采用逆转录聚合酶链反应和免疫印迹分析法，检测干细胞相关基因CD44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CD44的相对表达量高于原代贴壁细胞(P＜0.05)。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富集类肿瘤干细胞。该方法是分选和鉴定肿瘤干细胞的行之有效的方法。
목적：검측간세포상관인자CD44재위암세포주MKN45현부구체세포중적표체。방법：선택인위암세포주MKN45,재함유섬유생장인자-2급표피생장인자무혈청배양액중배양，관찰구체형성정황；채용역전록취합매련반응화면역인적분석법，검측간세포상관기인CD44재구체세포급원대첩벽세포량조세포중적표체차이。결과：위암세포주MKN45재무혈청배양조건하능형성현부구체,구체세포중CD44적상대표체량고우원대첩벽세포(P＜0.05)。결론：현부구배양법재위암세포주MKN45중배양형성적구체세포부집류종류간세포。해방법시분선화감정종류간세포적행지유효적방법。
Objective:To detect the expression of stem-cell related factor CD44 in spheroid body-forming cells of gastric cancer cell line MKN45. Methods: Gastric cancer cell line MKN45 were used to culture spheroid bodies in nonad-herent condition in a serum-free medium (SFM), supplemented with Epidermal Growth Factor (EGF) and Basic Fibroblast Growth Factor(bFGF). By using RT-PCR and western blot analysis, the expression levels of the embryonic stem cell-related gene CD44 were studied. Results: MKN45 cells formed spheroid bodies in nonadherent condition in a serum-free medium were observed, and the spheroid body-forming cells showed higher expressions of the embryonic stem cell-related gene CD44. Conclusion: Spheroid body-forming cells developed from human gastric cancer cell line MKN45 in nonadherent condition in a serum-free medium supplemented with Epidermal Growth Factor (EGF) and Basic Fibroblast Growth Factor (bFGF) were enriched in cancer stem-like cells. Spheroid body formation assay may be a practical and effective approach to isolate and identify CSCs.