γ射线辐照后NK92细胞对HOS骨肉瘤细胞杀伤作用的体外研究  

Ex-vivo study of the cytolytic effect of γ-ray-irradiated NK92 cells against HOS osteosarcoma cells

作　　者：胡家朗[1] 郑琼[1] 叶哲伟[2] 勘武生[1] 

机构地区：[1]湖北省武汉市普爱医院,430033 [2]华中科技大学附属协和医院,武汉市430022

出　　处：《中国矫形外科杂志》2013年第21期2212-2217,共6页Orthopedic Journal of China

基　　金：湖北省生物靶向治疗研究重点实验室开放课题资助项目(编号:2007B08)

摘　　要：[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0，3，10Gy，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2ng／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1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）。2ng／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 cytotoxic activity of T- ray-irradiated NK92 cells against HOS osteosarcoma cells and to investigate the cytolytic effect of rhlL - 12 - stimulated effector cells against the target cells. [ Methods] NK92 cells were exposed to different doses of radiation, orstimulated by 2ng/mLof rhlL - 12 for approximately 24h. We measured the cytotoxic activity of NK92 cells against targets by using the MTT colorimetric method and the quantity of Fas present on the surface of the osteosarcoma cells by using flow cytometry. [ Results ] The cytotoxic activity of NK92 cells against HOS was approximately 4. 3% -48%. In contrast to the control group, the cytotoxic activity of the NK92 cells irradiated against the target cells was only minimally greater, but that of the rhlL - 12 - stimulated NK92 cells was significantly higher. [ Conclusion] As T - ray - irradiated NK92 cells have excellent eytotoxic effecton HOS cells and reduce their proliferation activity significantly, they could be ideal for adoptive immunotherapy for osteosarcoma. As rhIL - 12 increases the cytotoxic activity of NK92 cells significantly, it has a good upregulating effect.

关 键 词：NK细胞系 过继免疫治疗 骨肉瘤 IL-12 

分 类 号：R738.1[医药卫生—肿瘤]