摘要
目的 观察应用反义RNA阻断胚胎横纹肌肉瘤细胞内源性IL 15表达后 ,肿瘤细胞对NK细胞杀伤敏感性及其体内、外增殖的变化。方法 应用流式细胞技术检测RD细胞及转染IL 15反义RNA的RD 10细胞表面MHCⅠ类 (包括HLA ABC重链及β2M链 )分子的表达 ;应用LDH释放法检测NK细胞对RD和RD 10细胞的细胞毒活性 ;通过对转染细胞的细胞周期分析以及接种到裸鼠体内来评价IL 15对肿瘤细胞增殖的影响。结果 ①阻断IL 15的表达可使HLA ABC分子阳性细胞百分率从 19.5 %升至 95 .5 % ;β2M分子阳性细胞百分率从 31%升至 90 .6 %。由于MHCⅠ类分子表达的增高 ,RD 10细胞对NK细胞的杀伤敏感性低于RD细胞 (P <0 .0 1)。②RD 10细胞的增殖减慢 ,阻滞在G0 /G1期。动物实验的结果也表明RD 10细胞在体内的致瘤性低于RD细胞。结论 阻断RD细胞内源性IL 15的表达可抑制该细胞的增殖 ,从而提示IL
Objective To investigate the variation of cytotoxicity to NK cells and the proliferation in vitro and in vivo of rhabdomyosarcoma cells after blocking the endogenous expression of IL 15 using anti sense RNA. Methods The expression of MHC class Ⅰ molecules including HLA ABC and β2M chains on RD and RD 10 cells were detected by flow cytometry. NK cytotoxicity was observed with LDH release methods. The cell cycle of the transfected cells was evaluated by flow cytometry. The proliferation was determined by means of in nude mice heterograft model in vivo . Results The results of flow cytometry showed RD cells had low level of HLA ABC and β2M molecules and the rate of positive cells is 19.5% and 31% respectively, whereas the rate of positive cells in RD 10 were elevated up to 95.5% and 90.6%. The NK cytotoxicity was inhibited in RD 10 cells with high expression of MHC class Ⅰ molecules. Flow cytometric analysis demonstrated an increased cell number of G 0/G 1 phase and RD 10 cells had low tumorigenicity in nude mice. Conclusion Suppress the proliferation of RD cells by blocking the expression of IL 15, which indicates the various role of IL 15 on tumors.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2002年第4期243-245,249,共4页
Immunological Journal
基金
国家自然科学基金资助项目 (39970 82 6 )