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目的研究CD133在人喉癌细胞系Hep-2中的表达,观察纯化的CD133+肿瘤细胞体外生长特性,确定喉癌肿瘤起始细胞的表面标志。方法免疫荧光细胞化学技术及流式细胞仪检测喉癌Hep-2细胞系中的CD133表达,免疫磁珠分选技术纯化CD133+肿瘤细胞,体外培养并观察其增殖及分化能力。结果喉癌Hep-2细胞系中有3·22%的微量细胞CD133呈阳性表达,免疫磁珠富集的CD133+肿瘤细胞在无血清培养基中3、5、7d的吸光度分别为0.320、0.370、0.558,均高于相同条件下未分选细胞和CD133-细胞;CD133+在培养体系中的比例逐日下降,至培养的第12天,由第1天的90.88%下降至4.53%。结论喉癌Hep-2细胞系中,CD133+癌细胞有比其他细胞亚群强的体外分化和增殖能力,CD133是肿瘤起始细胞的标志之一。
Objective To study the expression of CD133 in human laryngeal carcinoma cell line Hep-2 and to observe the growth characteristics of purified CD133 + tumor cells in vitro and to determine the surface markers of laryngeal cancer cell lines. Methods The expression of CD133 in laryngeal carcinoma Hep-2 cell line was detected by immunofluorescence cytochemistry and flow cytometry. The CD133 + tumor cells were purified by magnetic activated cell sorting and cultured in vitro. The proliferation and differentiation of CD133 tumor cells were observed. Results In the laryngeal carcinoma Hep-2 cell line, 3.22% of microtubules were positive for CD133. The absorbance of CD133 + tumor cells enriched in immunomagnetic beads in serum-free medium at 3, 5 and 7 days were 0.320 and 0.370, 0.558, all of which were higher than that under the same conditions. The proportion of CD133 + in the culture system decreased day by day and decreased from 90.88% on the first day to 4.53% on the 12th day of culture. Conclusion CD133 cancer cells have stronger differentiation and proliferation ability than other cell subsets in laryngeal cancer Hep-2 cell line. CD133 is one of the markers of tumor initiating cells.