Nanog与CD44在胃癌细胞株MKN45球体细胞中的表达及其意义

目的：检测与干细胞相关的2种因子胚胎干细胞相关转录因子4（Nanog）和分化抗原簇44（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，在含有表皮生长因子（EGF）及成纤维生长因子（bFGF）无血清培养液中培养，观察球体形成情况；采用免疫印迹（Western blot）、免疫荧光与实时定量聚合酶链反应（qRT-PCR）检测干细胞相关基因Nanog和CD44在球体细胞及原代贴壁细胞中的表达差异。结果胃癌细胞株MKN45在无血清培养条件下能形成悬浮球体，球体细胞中的Nanog和CD44的mRNA相对表达量分别为2.34±0.22和1.18±0.04，明显高于贴...

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Bibliographic Details
Published in天津医药 Vol. 43; no. 1; pp. 30 - 33
Main Author 刘建明 周友浪 马利林 徐骏飞 章建国
Format Journal Article
LanguageChinese
Published 南通大学附属医院普外科 邮编226001 2015
Subjects
CD44
Nanog
悬浮培养法
抗原
肿瘤干细胞
胃肿瘤
neoplastic stem cells
抗原, CD44
antigens,CD44
悬浮培养法
胃肿瘤
Nanog
stomach neoplasms
suspension culture
肿瘤干细胞
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ISSN0253-9896
DOI10.3969/j.issn.0253-9896.2015.01.008

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Summary:目的：检测与干细胞相关的2种因子胚胎干细胞相关转录因子4（Nanog）和分化抗原簇44（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，在含有表皮生长因子（EGF）及成纤维生长因子（bFGF）无血清培养液中培养，观察球体形成情况；采用免疫印迹（Western blot）、免疫荧光与实时定量聚合酶链反应（qRT-PCR）检测干细胞相关基因Nanog和CD44在球体细胞及原代贴壁细胞中的表达差异。结果胃癌细胞株MKN45在无血清培养条件下能形成悬浮球体，球体细胞中的Nanog和CD44的mRNA相对表达量分别为2.34±0.22和1.18±0.04，明显高于贴壁细胞中的Nanog和CD44的mRNA相对表达量1.00±0.00和1.00±0.05；球体细胞中的Nanog和CD44的蛋白质相对表达量分别为0.18±0.02和0.24±0.04，明显高于贴壁细胞中的相对表达量0.07±0.02和0.18±0.01（P＜0.05）。球体细胞中Nanog主要分布在细胞核周围及细胞质中，而CD44主要在细胞膜上表达，且同一个球体细胞中的Nanog与CD44大多同时表达。结论胃癌细胞株MKN45在无血清培养条件下形成的球体细胞具有肿瘤干细胞的特性。Nanog/CD44共表达细胞可能是胃癌干细胞的一种表型。
Bibliography:Nanog;stomach neoplasms;neoplastic stem cells;suspension culture;Nanog
Objective To detect the expression of stem-cell related factors Nanog and CD44 in spheroid body-form？ing cells of gastric cancer cell line MKN-45. Methods The gastric cancer cell line MKN-45 was used to culture spheroid bodies in non-adherent condition in a serum-free medium supplemented with epidermal growth factor （EGF） and basic fibro？blast growth factor （bFGF）. Using Western blot analysis, immunofluorescence staining and quantitative real time polymerase chain reaction（qRT-PCR）, the expression levels of stem cell-related genes Nanog and CD44 were studied. Results In this study, we observed that MKN45 cells formed spheroid bodies in non-adherent condition in a serum-free medium, and the levels of Nanog and CD44 mRNA expression in spheroid body-forming cells were 2.34 ± 0.22 and 1.18 ± 0.04,respectively, which were higher than those in parental cells （1.00±0.00 and 1.00±0.05）. The levels of Nanog and CD44 protein expression in spheroid bo
ISSN:0253-9896
DOI:10.3969/j.issn.0253-9896.2015.01.008