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目的:建立体外扩增人脐带血来源的NK细胞的方法,并研究其对多株人食管癌细胞系和食管癌患者原代肿瘤细胞的杀伤作用。方法:分离人脐带血单个核细胞,加入放射线灭活的人白血病K562细胞及IL-2、IL-15、IL-18细胞因子体外扩增培养2周,诱导NK细胞。采用流式细胞术检测NK细胞的扩增效果及其产生的IFN-γ等细胞因子杀伤靶细胞K562的能力,并检测体外扩增的人脐带血来源NK细胞对多种食管癌细胞系和食管癌患者原代肿瘤细胞凋亡的影响。结果:利用放射线灭活的K562细胞联合多种细胞因子,可以有效的扩增人脐带血来源的NK细胞。体外培养2周后NK细胞比率可达80%左右,并对靶细胞K562有明显促凋亡作用。此外,体外扩增的人脐带血来源NK细胞能够促进6株人食管癌细胞系和食管癌患者原代肿瘤细胞的凋亡。结论:本研究建立了体外扩增人脐带血NK细胞的方法,并证实这些NK细胞对食管癌细胞的促凋亡作用,可能为食管癌的免疫治疗提供新的手段。
OBJECTIVE: To establish a method to expand human umbilical cord blood-derived NK cells in vitro and study its killing effect on human esophageal cancer cell lines and primary tumor cells in patients with esophageal cancer. Methods: Human umbilical cord blood mononuclear cells were isolated and cultured. K562 cells and IL-2, IL-15 and IL-18 cytokines were inactivated by radiation in vitro. Flow cytometry was used to detect the proliferation of NK cells and the ability of cytokines such as IFN-γ to kill target cell K562. Flow cytometry was used to detect the proliferation of human umbilical cord blood-derived NK cells in vitro and in vitro Effect of primary tumor cell apoptosis in cancer patients. Results: Radiation-inactivated K562 cells combined with various cytokines could effectively proliferate human umbilical cord blood-derived NK cells. After 2 weeks of in vitro culture, the ratio of NK cells reached about 80%, and the apoptosis of K562 cells was obviously promoted. In addition, in vitro expanded human umbilical cord blood-derived NK cells can promote the apoptosis of primary human esophageal cancer cell lines and esophageal cancer cells in 6 human esophageal cancer cell lines. Conclusion: This study established a method of expanding human umbilical cord blood NK cells in vitro and confirmed that these NK cells could promote the apoptosis of esophageal cancer cells, which may provide a new method for immunotherapy of esophageal cancer.