摘要
目的研究苦参碱(Matrine)作用前后白血病KG1a细胞对NK细胞杀伤细胞敏感性的变化,并初步探讨其机制。方法应用CCK-8法检测苦参碱对KG1a细胞50%抑制量IC50值,以此量的苦参碱作用于KG1a细胞,LDH释放法检测作用前后对NK细胞的杀伤敏感性。RT-PCR检测KG1a细胞NKG2D配体(MICA、MICB、ULBP1、ULBP2、ULBP3)五种基因的表达,流式细胞仪检测作用前后KG1a细胞表面NKG2D配体和HLA-Ⅰ类分子表达变化。结果苦参碱作用前后在效靶比为5:1、10:1、20:1时对NK细胞杀伤敏感性差异均有统计学意义(P<0.05)。KG1a细胞在mRNA水平五种基因均表达,苦参碱作用前KG1a细胞表面NKG2D各配体几乎不表达,作用后各配体显著升高,与作用前相比差异有统计学意义(P<0.05),HLA-Ⅰ类分子无明显变化(P>0.05)。结论苦参碱能提高KG1a细胞对NK细胞的杀伤敏感性,其机制可能与苦参碱提高KG1a细胞表面NKG2D配体的表达有关。
Objective To study the effects of NK cell cytotoxicity against leukemic cell lines KG1a treated with Matrine and to preliminarily explore their molecular mechanisms. Methods The IC50 of KG1a treated with Matrine for 48h was determined with the cck-8 assay and this level of Matrine was chosen to treat KG1a cells. Cytotoxicity of NK cells isolated from 6 healthy volunteers against KG1a cell were ana lyzed by LDH releasing assay at different effeetor-to target cell ratios（E：T） before and after treated with Matrine. The genes expression of NKG2D ligands（MICA, MICB, ULBP1, ULBP2, ULBP3） of KG1 a cells by RT-PCR, the expression of NKG2D ligands and HLA-class I molecules were assayed by flow cytometery before and after treated with Matrine. Results The cytotoxicity of NK cells against KG1a cells treated with Matrine was enhanced at 5： 1;10： 1;20：1 of effect-to-target（P〈0. 05）. The genes of NKG2D ligands were positively detected in KGla cells at mRNA level, but there were hardly detected at the pro tein level. However, the expression of NKG2D ligands of KG1a cells were up-regulated after treated with Matrine（P〈0. 05）. There was no significant difference of expression HLA-class I molecules between before and after treated with Matrine（P〉0. 05）. Conclusion The Matrine could obviously up-regulate the NKG2D ligands expression in KGla cells and thus enhance the cytotoxicity of NK cell against the KGla cells.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2009年第4期298-301,共4页
Cancer Research on Prevention and Treatment
基金
广州市科技计划资助项目(2005Z3-E0551)
