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Identification of cancer stem-like CD44+ cells in human nasopharyngeal carcinoma cell line.

机译:鉴定人鼻咽癌细胞系中的癌干样CD44 +细胞。

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BACKGROUND AND AIMS: Recent studies suggest that cancer stem cells (CSC) may be responsible for tumorigenesis and contribute to some individuals' resistance to cancer therapy. Although research is rapidly advancing in this field, to our knowledge there are few published reports about the CSC in human nasopharyngeal carcinoma (NPC). We undertook this study to separate, expand, and explore the biological features of CD44+ stem-like cancer cells from the human NPC SUNE-1 5-8F cell line. METHODS: Immunocytochemistry and flow cytometry were used to detect the expression of CD44 in SUNE-1 5-8F. Fluorescence-activated cell sorting was applied to purify CD44+ cells. MTT assay or clone formation assay was used to detect the differences of CD44+ and CD44- cells in proliferation, differentiation, radiosensitivity and chemosensitivity in vitro. The expression of stem cell markers Oct-4 and Bmi-1 was examined by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: CD44 was positively expressed in approximately 52.5% of NPC SUNE-1 5-8F cell line. Regardless of serum-free medium and serum medium culture conditions, freshly sorted CD44+ cells showed stronger proliferative capacity than CD44- and unsorted cells. The expression levels of Bmi-1 and Oct-4 mRNA in CD44+ cells were significantly higher than CD44- cells. After 2 Gy radiation, the average clone formation efficiency for CD44+ and CD44- cells was 22.17 +/- 6.65% and 11.50 +/- 5.00%, respectively (p <0.05). After cisplatin and docetaxel treatment with the same drug concentration, CD44+ cells showed a higher survival rate compared with CD44- cells. CONCLUSIONS: CD44+ cells have the biological characteristics of tumor stem cell and may be assumed as one of the markers of NPC tumor stem cells.
机译:背景与目的:最近的研究表明,癌症干细胞(CSC)可能与肿瘤发生有关,并导致某些人对癌症治疗产生抗性。尽管该领域的研究正在迅速发展,但据我们所知,关于人鼻咽癌(NPC)中CSC的报道很少。我们进行了这项研究,以分离,扩增和探索来自人NPC SUNE-1 5-8F细胞系的CD44 +干样癌细胞的生物学特性。方法:采用免疫细胞化学和流式细胞仪检测SUNE-1 5-8F中CD44的表达。荧光激活细胞分选用于纯化CD44 +细胞。使用MTT测定或克隆形成测定来检测CD44 +和CD44-细胞在体外增殖,分化,放射敏感性和化学敏感性方面的差异。通过逆转录聚合酶链反应(RT-PCR)检测干细胞标志物Oct-4和Bmi-1的表达。结果:CD44在约52.5%的NPC SUNE-1 5-8F细胞系中阳性表达。无论无血清培养基和血清培养基培养条件如何,新鲜分选的CD44 +细胞均比CD44-和未分选的细胞具有更强的增殖能力。 CD44 +细胞中Bmi-1和Oct-4 mRNA的表达水平明显高于CD44-细胞。 2 Gy辐射后,CD44 +和CD44-细胞的平均克隆形成效率分别为22.17 +/- 6.65%和11.50 +/- 5.00%(p <0.05)。用相同的药物浓度处理顺铂和多西他赛后,与CD44-细胞相比,CD44 +细胞显示出更高的存活率。结论:CD44 +细胞具有肿瘤干细胞的生物学特性,可作为NPC肿瘤干细胞的标志物之一。

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