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目的探讨HLAⅠ类分子及MHCⅠ类链相关分子(MICA/MICB)在人鼻咽癌细胞株CNE2及多药耐药细胞株CNE2/DDP的表达及其对NK细胞杀伤活性的影响。方法流式细胞仪检测CNE2和CNE2/DDP细胞表面HLAⅠ类分子和MICA/MICB的表达情况;LDH释放法测定3例健康者的NK细胞在不同效靶比时对CNE2、CNE2/DDP细胞的杀伤活性;效靶比10∶1时,用抗HLAⅠ类分子单抗(W6/32)和抗MICA/MICB单抗(BAMO-1)分别封闭HLAⅠ类分子和MICA/MICB,观察NK细胞杀伤CNE2、CNE2/DDP细胞活性的变化。结果CNE2/DDP细胞表面HLAⅠ类分子和MICA/MICB的表达均较CNE2细胞明显减少(P<0.01)。NK细胞对CNE2、CNE2/DDP细胞的杀伤活性在效靶比5∶1时分别是(9.37±2.14)%、(4.37±0·63)%;效靶比10∶1时分别是(27.14±1.82)%、(15.79±2.87)%;效靶比20∶1时分别是(36.40±4.28)%、(26.20±4·18)%;效靶比301时分别是(54.67±2.80)%、(40.29±2.73)%。各效靶比时NK细胞对CNE2、CNE2/DDP细胞的杀伤活性与HLAⅠ类分子和MICA/MICB相关,NK细胞对CNE2/DDP细胞的杀伤活性明显低于CNE2细胞(P<0.01)。效靶比10∶1时,W6/32明显增强NK细胞对CNE2、CNE2/DDP细胞的杀伤活性(P<0.01),BAMO-1明显抑制NK细胞对CNE2、CNE2/DDP细胞的杀伤活性(P<0.01)。结论HLAⅠ类分子和MICA/MICB在CNE2、CNE2/DDP的表达差异影响着NK细胞的杀伤活性,MICA/MICB在多药耐药肿瘤细胞的低表达导致耐药肿瘤细胞对NK细胞杀伤敏感性下降。
Objective To investigate the expression of HLA class I and MICA / MICB on human nasopharyngeal carcinoma cell line CNE2 and multidrug-resistant cell line CNE2 / DDP and its effect on cytotoxicity of NK cells. Methods The expression of HLA class I molecules and MICA / MICB on the surface of CNE2 and CNE2 / DDP cells were detected by flow cytometry. The cytotoxicity of NK cells on CNE2 and CNE2 / DDP cells was measured by LDH release assay at different effective target ratios Activity and HLA-class I molecules and MICA / MICB were respectively blocked by anti-HLA class I monoclonal antibody (W6 / 32) and anti-MICA / MICB monoclonal antibody (BAMO-1) Changes in CNE2 / DDP cell activity. Results The expression of HLA class I molecules and MICA / MICB on CNE2 / DDP cells was significantly lower than that on CNE2 cells (P <0.01). The killing activity of NK cells on CNE2 and CNE2 / DDP cells was (9.37 ± 2.14)% and (4.37 ± 0.63)%, respectively, when the effective target ratio was 5:1; 1.82%, and (15.79 ± 2.87)%, respectively. The effective target ratios were (36.40 ± 4.28)% and (26.20 ± 4.18)% at 20:1 respectively; (40.29 ± 2.73)%. The killing activity of NK cells on CNE2 and CNE2 / DDP cells was correlated with HLA class I molecules and MICA / MICB at various effective target ratios. The cytotoxic activity of NK cells to CNE2 / DDP cells was significantly lower than that of CNE2 cells (P <0.01). The ratio of NKT to CNE2 and CNE2 / DDP cells was significantly increased by W6 / 32 (P <0.01), while BAMO-1 significantly inhibited the cytotoxicity of NK cells to CNE2 and CNE2 / DDP cells <0.01). Conclusion The expression of HLA class I molecules and MICA / MICB in CNE2 and CNE2 / DDP affect the killing activity of NK cells. The low expression of MICA / MICB in multidrug-resistant tumor cells results in the decrease of sensitivity of drug-resistant tumor cells to NK cell killing .