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[目的]研究NK92细胞经γ射线辐照后的增殖活性及对HOS骨肉瘤细胞的体外杀伤效应变化;观察rhIL-12对γ射线辐照后NK92细胞的胞毒杀伤效力的影响。[方法]应用60Coγ射线发生器对NK92进行照射处理,照射剂量分别为0,3,10 Gy,靶细胞为K562细胞及HOS细胞,其中K562细胞为阳性对照。MTT法检测NK92细胞经射线受照后的增殖能力及不同处理条件下NK92细胞对靶细胞的杀伤活性。将2 ng/ml rhIL-12加入预照射处理后的NK92培养基中孵育过夜,检测rhIL-12对γ射线辐照后NK92细胞的胞毒杀伤效力的影响,流式细胞术检测细胞表面Fas表达的变化。[结果]随着效靶比梯度的变化,NK92对HOS的杀伤率在4.3%~48.0%之间。与未照射组相比,经射线处理后的NK92细胞对靶细胞的杀伤率差异无统计学意义(P>0.05)。2 ng/ml rhIL-12能明显提高NK92细胞及γ射线辐照后的NK92细胞的体外杀伤作用(P<0.05)。[结论]γ射线辐照后的NK92细胞对HOS细胞具有良好的杀伤活性,同时其自身增殖活性明显降低,可作为骨肉瘤过继免疫治疗的理想效应细胞。此外,rhIL-12能明显提高NK92细胞对靶细胞的杀伤活性,起到良好正向调节作用。
[Objective] To study the proliferation activity of NK92 cells after γ-ray irradiation and the killing effect on osteosarcoma cells in vitro and the effect of rhIL-12 on cytotoxicity of NK92 cells after γ-ray irradiation. [Methods] NK92 was irradiated with 60Co γ-ray generator at doses of 0, 3 and 10 Gy, respectively. The target cells were K562 cells and HOS cells, of which K562 cells were positive control. The proliferation of NK92 cells irradiated by irradiation and the killing activity of NK92 cells on target cells under different treatment conditions were detected by MTT assay. After 2 ng / ml rhIL-12 was added to the pre-irradiated NK92 medium for incubation overnight, the effect of rhIL-12 on cytotoxicity of NK92 cells after γ-ray irradiation was examined. Fas expression on the cell surface was detected by flow cytometry The change. [Result] The killing rate of NK92 to HOS ranged from 4.3% to 48.0% with the change of target-specific gradient. Compared with the non-irradiated group, there was no significant difference in the killing rate of target cells between irradiated and irradiated NK92 cells (P> 0.05). 2 ng / ml rhIL-12 could significantly enhance the killing effect of NK92 cells and γ-ray irradiated NK92 cells in vitro (P <0.05). [Conclusion] The γ-ray irradiation of NK92 cells have good cytotoxic activity on HOS cells, meanwhile, their proliferation activity is obviously decreased, which can be used as ideal effector cells for adoptive immunotherapy of osteosarcoma. In addition, rhIL-12 can significantly improve the killing activity of NK92 cells on target cells, play a good positive regulatory role.