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Significantly increased expression of OCT4 and ABCG2 in spheroid body-forming cells of the human gastric cancer MKN-45 cell line

机译:OCT4和ABCG2在人胃癌MKN-45细胞系球状体形成细胞中的表达显着增加

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摘要

The cancer stem cell (CSC) theory hypothesizes that CSCs are the cause of tumor formation, recurrence and metastasis. Key to the study of CSCs is their isolation and identification. The present study investigated whether spheroid body-forming cells in the human gastric cancer (GC) MKN-45 cell line are enriched for CSC properties, and also assessed the expression of the candidate CSC markers, octamer-binding transcription factor-4 (OCT4) and adenosine triphosphate-binding cassette transporter G2 (ABCG2) in the MKN-45 spheroid body cells. The MKN-45 cells were plated in a stem cell-conditioned culture system to allow for spheroid body formation. The expression levels of OCT4 and ABCG2 in the spheroid body cells were assessed by qPCR, western blot analysis and immunofluorescence staining, while the tumorigenicity of the spheroid body-forming cells was assessed by in vivo xenograft studies in nude mice. The MKN-45 cells were able to form spheroid bodies when cultured in stem cell-conditioned medium. The spheroid body-forming cells showed a significantly higher (P<0.01) expression of OCT4 and ABCG2 compared with the parental cells. These data suggest that the spheroid body cells from the MKN-45 GC cell line cultured in stem cell-conditioned medium possessed gastric CSC properties. The co-expression of OCT4 and ABCG2 by these cells may represent the presence of a subpopulation of gastric CSCs.
机译:癌症干细胞(CSC)理论假设,CSC是肿瘤形成,复发和转移的原因。研究CSC的关键是其隔离和识别。本研究调查了人胃癌(GC)MKN-45细胞系中的球状体形成细胞是否具有丰富的CSC特性,并评估了候选CSC标志物,八聚物结合转录因子4(OCT4)的表达。 MKN-45球体细胞中的三磷酸腺苷结合盒转运蛋白G2(ABCG2)。将MKN-45细胞接种在干细胞条件培养系统中,以形成球体。通过qPCR,蛋白质印迹分析和免疫荧光染色评估球状体细胞中OCT4和ABCG2的表达水平,同时通过体内裸鼠体内异种移植研究评估球状体形成细胞的致瘤性。在干细胞条件培养基中培养时,MKN-45细胞能够形成球体。与亲代细胞相比,球状体形成细胞显示OCT4和ABCG2的表达显着更高(P <0.01)。这些数据表明,在干细胞条件培养基中培养的来自MKN-45 GC细胞系的球状体细胞具有胃CSC特性。这些细胞对OCT4和ABCG2的共表达可能代表了胃CSC亚群的存在。

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