【摘要】：目的:建立體外擴(kuò)增人臍帶血來(lái)源的NK細(xì)胞的方法,并研究其對(duì)多株人食管癌細(xì)胞系和食管癌患者原代腫瘤細(xì)胞的殺傷作用。方法:分離人臍帶血單個(gè)核細(xì)胞,加入放射線滅活的人白血病K562細(xì)胞及IL-2、IL-15、IL-18細(xì)胞因子體外擴(kuò)增培養(yǎng)2周,誘導(dǎo)NK細(xì)胞。采用流式細(xì)胞術(shù)檢測(cè)NK細(xì)胞的擴(kuò)增效果及其產(chǎn)生的IFN-γ等細(xì)胞因子殺傷靶細(xì)胞K562的能力,并檢測(cè)體外擴(kuò)增的人臍帶血來(lái)源NK細(xì)胞對(duì)多種食管癌細(xì)胞系和食管癌患者原代腫瘤細(xì)胞凋亡的影響。結(jié)果:利用放射線滅活的K562細(xì)胞聯(lián)合多種細(xì)胞因子,可以有效的擴(kuò)增人臍帶血來(lái)源的NK細(xì)胞。體外培養(yǎng)2周后NK細(xì)胞比率可達(dá)80%左右,并對(duì)靶細(xì)胞K562有明顯促凋亡作用。此外,體外擴(kuò)增的人臍帶血來(lái)源NK細(xì)胞能夠促進(jìn)6株人食管癌細(xì)胞系和食管癌患者原代腫瘤細(xì)胞的凋亡。結(jié)論:本研究建立了體外擴(kuò)增人臍帶血NK細(xì)胞的方法,并證實(shí)這些NK細(xì)胞對(duì)食管癌細(xì)胞的促凋亡作用,可能為食管癌的免疫治療提供新的手段。
[Abstract]:Aim: to establish a method to amplify NK cells derived from human umbilical cord blood in vitro and to study its killing effect on human esophageal cancer cell lines and primary tumor cells from esophageal cancer patients. Methods: human umbilical cord blood mononuclear cells were isolated and cultured in vitro with radiation-inactivated human leukemia K562 cells and IL-2,IL-15,IL-18 cytokines for 2 weeks to induce NK cells. Flow cytometry was used to detect the amplification effect of NK cells and the ability of IFN- 緯 and other cytokines to kill K562 target cells. The effects of NK cells derived from human umbilical cord blood on apoptosis of various esophageal cancer cell lines and primary tumor cells from patients with esophageal carcinoma were examined. Results: NK cells derived from human umbilical cord blood could be effectively amplified by radiation inactivated K562 cells combined with many cytokines. After 2 weeks of culture in vitro, the ratio of NK cells was about 80%, and the target cell K562 could induce apoptosis. In addition, NK cells derived from human umbilical cord blood in vitro could promote apoptosis of 6 human esophageal cancer cell lines and primary tumor cells of esophageal cancer patients. Conclusion: this study established a method to amplify human umbilical cord blood NK cells in vitro, and confirmed that these NK cells can promote apoptosis of esophageal cancer cells, which may provide a new method for immunotherapy of esophageal cancer.
【作者單位】： 鄭州大學(xué)第一附屬醫(yī)院生物細(xì)胞治療中心;鄭州大學(xué)第一臨床學(xué)院;鄭州大學(xué)生命科學(xué)學(xué)院;
【基金】：河南省基礎(chǔ)與前沿技術(shù)研究計(jì)劃基金資助項(xiàng)目(No.142300410387.0)~~
【分類號(hào)】：R735.1

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