【摘要】：目的:建立體外擴增人臍帶血來源的NK細胞的方法,并研究其對多株人食管癌細胞系和食管癌患者原代腫瘤細胞的殺傷作用。方法:分離人臍帶血單個核細胞,加入放射線滅活的人白血病K562細胞及IL-2、IL-15、IL-18細胞因子體外擴增培養(yǎng)2周,誘導NK細胞。采用流式細胞術檢測NK細胞的擴增效果及其產(chǎn)生的IFN-γ等細胞因子殺傷靶細胞K562的能力,并檢測體外擴增的人臍帶血來源NK細胞對多種食管癌細胞系和食管癌患者原代腫瘤細胞凋亡的影響。結果:利用放射線滅活的K562細胞聯(lián)合多種細胞因子,可以有效的擴增人臍帶血來源的NK細胞。體外培養(yǎng)2周后NK細胞比率可達80%左右,并對靶細胞K562有明顯促凋亡作用。此外,體外擴增的人臍帶血來源NK細胞能夠促進6株人食管癌細胞系和食管癌患者原代腫瘤細胞的凋亡。結論:本研究建立了體外擴增人臍帶血NK細胞的方法,并證實這些NK細胞對食管癌細胞的促凋亡作用,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】： 鄭州大學第一附屬醫(yī)院生物細胞治療中心;鄭州大學第一臨床學院;鄭州大學生命科學學院;
【基金】：河南省基礎與前沿技術研究計劃基金資助項目(No.142300410387.0)~~
【分類號】：R735.1

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