耐地塞米松B细胞淋巴瘤细胞逃逸同种异体NK细胞的杀伤及其机制  

Mechanism of dexamethasone-resistant B-cell lymphoma cells escaping cytotoxicity of allogeneic NK cells

作　　者：谢宁 周健 宋永平 高全立 张艳莉 符粤文 房佰俊 魏旭东 

机构地区：[1]郑州大学附属肿瘤医院血液科；河南郑州450003

出　　处：《中国肿瘤生物治疗杂志》2012年第3期247-251,共5页Chinese Journal of Cancer Biotherapy

基　　金：河南省科技厅科研项目(No.0611042000)~~

摘　　要：目的:观察地塞米松耐药的人B细胞淋巴瘤细胞系对NK细胞杀伤敏感性的变化,并探讨其作用机制。方法:20μg/ml地塞米松(dexamethasone,DXM)诱导B细胞淋巴瘤细胞系SU-DHL-4(简称SU细胞)发生耐药,建立多药耐药细胞系SU/DXM。流式细胞术分选健康人外周血NK细胞,流式细胞术检测效靶比20∶1时,NK细胞对SU和SU/DXM细胞的杀伤效应。实时定量PCR检测SU和SU/DXM细胞表面NK细胞活化性受体(soluble NK group 2 member D,NKG2D)配体基因[可溶性MHCⅠ类分子相关A/B(MHC classⅠchain-related molecules A/B,MICA/B)及人UL16结合蛋白(UL16 binding protein,ULBP)1、2、3]的表达。结果:成功建立多药耐药细胞系SU/DXM。与SU细胞相比,SU/DXM细胞对NK细胞杀伤的敏感性明显下降[SU细胞为(11.38±3.51)%,SU/DXM细胞为(3.57±4.22)%,P<0.05],细胞表面NKG2D配体基因MICA、MICB、ULBP2 mRNA表达量降低(SU细胞分别为1.014±0.121、1.009±0.092、0.993±0.108,SU/DXM细胞分别为0.017±0.006、0.682±0.063、0.773±0.066,P<0.05或P<0.01)。结论:地塞米松能诱导B细胞淋巴瘤SU细胞发生多药耐药,多药耐药SU/DXM细胞能够抵抗NK细胞的杀伤,其机制可能与NKG2D配体基因表达量下降有关。Objective： To observe the change of cytotoxicity of NK cells on dexamethasone-resistant B-cell lymphoma cells,and study its mechanism.Methods： 20 μg/ml dexamethasone（DXM） was added to induce DXM-resistance B lymphoma SU-DHL-4 cells（SU cells）.The multidrug resistance of SU cells was named SU-DXM cells.NK cells,sorted by flow cytometry was chosen as an effector,and the cytotoxicity of NK cells on SU and SU/DXM cells was analyzed through flow cytometry at E∶ T was 20∶ 1.Real-time PCR was used to detect the gene expression of MHC class Ⅰ chain-related molecules A/B（MICA/B）,UL16 binding protein（ULBP） 1,ULBP2 and ULBP3 in both SU and SU/DXM cells.Results： After DXM treatment,the SU /DXM cells were established as a multi-drug-resistant cell line.Compared to SU cells,the cytotoyxicity sensitivity of NK cells on SU/DXM cells was obviously down-regulated（[3.57 ± 4.22]% vs [11.38 ±3.51]%,P 0.05）,and the expressions of NKG2D ligand genes MICA,MICB and ULBP2 were statistically lower in SU/DXM cells（MICA： 0.017 ± 0.006,MICB： 0.682 ± 0.063,and ULBP： 20.773 ± 0.066） than those in SU cells（MICA： 1.014 ± 0.121,MICB： 1.009 ± 0.092,and ULBP2： 0.993 ± 0.108,P 0.05,P 0.01）.Conclusion： Devamethasone has the ability to induce SU cells to multi-drug-resistant cells,SU /DXM cells,which resist the cytotoxicity mediated by NK cells,and the low gene expression of NKG2D ligands could be one important cause.

关 键 词：B细胞淋巴瘤 地塞米松 NK细胞 多药耐药 杀伤 NKG2D 

分 类 号：R730.2[医药卫生—肿瘤] R730.3[医药卫生—临床医学]