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目的:研究NDV7793激活的NK细胞对人肝癌HepG2细胞的杀伤作用,并探讨可能的相关机制。方法:用免疫磁珠分离法分离人NK细胞,用乳酸脱氢酶释放法(LDH法)检测不同血凝单位(25.6Hu,51.2Hu,102.4Hu,204.8Hu,512.0Hu)的NDV7793刺激NK细胞后对人肝癌HepG2细胞的杀伤作用,分别用ELISA法和流式细胞术检测活化后的NK细胞分泌肿瘤坏死因子-α(TNF-α)的水平及细胞膜表面的TRAIL蛋白表达水平。结果:人外周血NK细胞分离纯度达(90.6±1.15)%;NDV7793能提高NK细胞对人肝癌HepG2细胞的杀伤作用,其杀伤效力(释放的LDH活力值)随着效靶细胞作用时间的增加及效靶细胞比的增大而增强;NDV7793能显著提高NK细胞TNF-α的分泌水平,其中204.8HuNDV7793刺激NK细胞12h后,TNF-α分泌水平达到高峰(0.184±0.01);102.4HuNDV7793刺激NK细胞16h后,NK细胞膜表面TRAIL表达达到高峰(22.77±1.8)%。结论:NDV7793能增强NK细胞对人肝癌HepG2细胞的杀伤作用,杀伤机制可能是和提高NK细胞分泌的TNF-α及上调细胞膜表面TRAIL蛋白有关。
OBJECTIVE: To study the killing effect of NDV7793-activated NK cells on HepG2 cells and to explore possible mechanisms. Methods: Human NK cells were isolated by immunomagnetic beads method and NDV7793 cells with different hemagglutination units (25.6Hu, 51.2Hu, 102.4Hu, 204.8Hu, 512.0Hu) were stimulated with lactate dehydrogenase (LDH) to stimulate NK cells After the killing effect on HepG2 cells, the levels of tumor necrosis factor-α (TNF-α) secreted by activated NK cells and the expression of TRAIL protein on the cell membrane were detected by ELISA and flow cytometry respectively. Results: The purity of NK cells in human peripheral blood was (90.6 ± 1.15)%. NDV7793 could enhance the killing effect of NK cells on HepG2 cells. The killing effect (release of LDH activity) NDV7793 could significantly increase the level of TNF-α secretion in NK cells. After NK cells were stimulated by 204.8HuNDV7793 for 12 hours, the secretion of TNF-α reached the peak (0.184 ± 0.01); 102.4HuNDV7793 stimulated NK After 16 hours, the expression of TRAIL on NK cell membrane peaked (22.77 ± 1.8)%. Conclusion: NDV7793 can enhance the killing effect of NK cells on HepG2 cells. The killing mechanism may be related to the increase of TNF-α secreted by NK cells and the up-regulation of TRAIL protein on cell surface.