目的：研究HLA-G反义寡核苷酸逆转肿瘤细胞免疫逃逸，提高免疫效应细胞识别杀伤活性的作用和机制。方法：采用反义核酸技术合成HLA-G反义寡核苷酸(ASODN)，硫代化修饰与脂质体形成复合物，转导入高表达HLA-G的绒毛膜癌细胞系JEG-3。采用RT-PCR方法检测HLA-G mRNA表达水平的变化；流式细胞术检测细胞表面HLA-G蛋白表达水平的变化； MTT法检测：NK杀伤活性、CD3AK杀伤、增殖活性的变化； ELISA方法探讨ASODN调节CD3AK细胞因子IFN-γ产生的变化。结果HLA-G ASODN可显著抑制HLA-G mRNA和蛋白水平的表达，逆转可溶型HLA-G分子对NK细胞杀伤活性的抑制作用；可部分逆转HLA-G分子对CD3AK细胞产生IFN-γ的抑制作用，并增强CD3AK的增殖活性，增强JEG-3细胞对CD3AK的杀伤敏感性。结论： HLA-G ASODN通过抑制肿瘤细胞HLA-G mRNA和蛋白水平的表达，增强NK，CD3AK的杀伤作用和机制，发挥逆转肿瘤细胞免疫逃逸作用。

Objective: To explore the mechanism of HLA-G antisense oligodeoxynucleotides (ASODN) on reversion of tumour immune evasion and enhancement of killer activity of immune effector cells. Methods: The expression of HLA-G in JEG-3 cell line was studied by RT-PCR and flow cytometry. MTT assay was used in our studies to detect the change of NK and CD3AK killer activity after ASODN treatment. The effect of IFN-γ secreted by CD3AK after coculture of tumour-lymphocytes was explored by ELISA.Results： HLA-G ASODN could obviously inhibit the expression of HLA-G mRNA and protein. The supernatant of JEG-3 treated by ASODN could reverse the inhibiton of soluble HLA-G molecules to NK killer activity. On the other hand, HLA-G ASODN also could reverse the inhibition of HLA-G to the production of IFN-γ secreted by CD3AK and enhanced the proliferation of CD3AK. Meanwhile, it also improved the susceptibility of JEG-3 to CD3AK.Conclusion:HLA-G ASODN could inhibits the expression of HLA-G mRNA and protein in tumour cells and partly restore the cytotoxicity of NK and CD3AK cells, and thereby partly reverse immune evasion of tumour cells caused by souble HLA-G molecules.

山东省自然科学基金资助项目（Y2001C08）