和厚朴酚提高人非霍奇金淋巴瘤Raji细胞对NK细胞杀伤敏感性的研究

摘要

Objective To observe the difference of cytotoxicity of natural killer(NK) cells against Raji cells before and after HNK treatment with HNK and to investigate the possible mechanism underlyings. Methods The inhibitive effect of HNK on Raji cells proliferation was detected by MTT assay. Cytotoxicity of NK cells against Raji cells before and after HNK treatment was determined by LDH releasing assay. The natural killer group 2 member D (NKG2D) ligands (MICA/B, ULBP1, ULBP2, and ULBP3) and human leukocyte antigen (HLA) class I molecules expression in Raji cells and cell cycle assay before and after HNK treatment was determined by flow cytoraetery assay, respectively. Results HNK significantly inhibited the Raji cells growth.The cytotoxicity of NK cells against Raji cells after HNK treatment was enhanced (P ＜ 0.05). NKG2D ligands (MICAB,ULBP2NULBP3) expression was incresed after HNK treatment (P ＜ 0.05). No significant difference was observed in HLA class I molecules and ULBP1 expression before and after HNK treatment (P ＞ 0.05). Raji cells was shown blocked at Go/G, phase after HNK treatment. Conclusion HNK up-regulated NKG2D ligands (MICABNULBP2,ULBP3) expression in Raji cells, and enhanced the cytotoxicity of NK cells against Raji cells contributing to high ability to kill tumor cells.%目的:探讨和厚朴酚(honokiol,HNK)作用前后人非霍奇金淋巴瘤Raji细胞对NK细胞杀伤敏感性的变化及其可能机制.方法:MTT法检测和厚朴酚对Raji细胞的增殖抑制作用.乳酸脱氢酶释放法检测HNK作用前后Raji细胞对NK细胞的杀伤敏感性.流式细胞仪检测HNK作用前后Raji细胞表面NKG2D配体(MICAB、ULBP1、ULBP2、ULBP3)和HLA-Ⅰ类分子的表达以及细胞周期变化.结果:HNK对Raji细胞的生长具有明显抑制作用.HNK作用后,Raji细胞对NK细胞的杀伤敏感性较HNK作用前显著增强(P＜0.05).HNK作用后,Raji细胞表面MICAB、ULBP2、ULBP3表达显著升高(P＜0.05),ULBP1和HLA-Ⅰ类分子无明显变化(P＞0.05),同时,细胞周期检测显示Raji细胞被阻滞在G0/G1期.结论:HNK能提高Raji细胞NKG2D配体(MICAB、ULBP1、ULBP3)的表达,从而使Raji细胞对NK细胞的杀伤敏感性增强,以达到杀伤肿瘤细胞的能力.