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目的分析人类胶质母细胞瘤(glioblastoma multiforme,GBM)细胞株U251和由此分离的胶质瘤干细胞中HMGA1的差异性表达。方法用MACS柱从U251中分离出表达表面标志物CD133的胶质瘤干细胞(glioblas-toma stem cells,GSCs),并用免疫荧光技术和流式细胞分析法对其进行分析。用实时反转录酶-聚合酶链反应技术(real-time PCR,RT-PCR)和蛋白质印迹技术分别探测到两类细胞的HMGA1基因在转录和翻译水平上表达的不同。结果从U251中分离出GSCs,U251中的GSCs约为0.32%。在GSCs中,HMGA1在转录和翻译水平上分别为U251蛋白的(6.13±0.25)倍和(2.75±0.99)倍。结论相比于U251,HMGA1在GSCs是过度表达的。HMGA1的过度表达可能与体内肿瘤干细胞的恶性扩增、侵袭和分化密切相关。HMGA1基因可望成为胶质母细胞瘤的一种生物标记物和治疗的靶向目标。
Objective To analyze the differential expression of HMGA1 in glioma stem cells of human glioblastoma multiforme (GBM) cell line U251. Methods Glioblas-derived stem cells (GSCs) expressing surface marker CD133 were isolated from U251 using MACS column and analyzed by immunofluorescence and flow cytometry. Real-time reverse transcriptase-polymerase chain reaction (real-time PCR, RT-PCR) and Western blotting were used to detect the difference of HMGA1 gene transcription and translation between the two cell types. Results GSCs were isolated from U251 and GSCs in U251 were approximately 0.32%. In GSCs, HMGA1 was (6.13 ± 0.25) fold and (2.75 ± 0.99) fold higher than the U251 protein at the transcriptional and translational levels, respectively. Conclusion HMGA1 is overexpressed in GSCs compared to U251. Overexpression of HMGA1 may be closely related to the malignant expansion, invasion and differentiation of cancer stem cells in vivo. The HMGA1 gene is expected to be a biomarker and target of therapy for glioblastoma.