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目的探讨IL-21对小泛素相关修饰蛋白特异性蛋白酶1(SENP1)表达的影响及其对NK细胞生物学特性的调控作用。方法以NK92细胞系为模型,采用慢病毒介导的Senp1基因干涉策略,抑制NK92细胞SENP1的表达。实时荧光定量PCR和Western印迹检测SENP1表达水平;利用CCK-8细胞增殖检测试剂盒测定细胞增殖活性;AnnexinⅤ-APC/PI标记检测细胞凋亡。将K562细胞与NK92细胞共培养,荧光素酶报告基因方法检测NK92细胞的杀伤活性。结果 IL-21处理上调NK92细胞SENP1的表达;慢病毒介导Senp1-shRNA转染显著降低NK92细胞内Senp1 mRNA和蛋白表达水平。Senp1干涉组NK92细胞增殖能力较对照组明显降低。同时,干涉Senp1能促进NK92细胞凋亡,但对NK92细胞杀伤K562活性无显著影响。结论 IL-21上调NK92细胞SENP1的表达;干涉Senp1能抑制NK92细胞增殖,促进细胞凋亡,同时影响穿孔素表达和对肿瘤细胞的杀伤活性。
Objective To investigate the effect of IL-21 on the expression of small ubiquitin-related modified protein-specific protease 1 (SENP1) and its regulation on the biological characteristics of NK cells. Methods Using the NK92 cell line as a model, the lentivirus mediated Senp1 gene interference strategy was used to inhibit the expression of SENP1 in NK92 cells. Real-time quantitative PCR and Western blot were used to detect the expression of SENP1. Cell proliferation was measured by CCK-8 cell proliferation assay kit. Apoptosis was detected by AnnexinⅤ-APC / PI. K562 cells were co-cultured with NK92 cells, and the killing activity of NK92 cells was detected by luciferase reporter assay. Results IL-21 treatment up-regulated the expression of SENP1 in NK92 cells. Lentivirus-mediated Senp1-shRNA transfection significantly reduced the expression of Senp1 mRNA and protein in NK92 cells. The proliferation of NK92 cells in the Senp1-treated group was significantly lower than that in the control group. At the same time, interference with Senp1 could promote the apoptosis of NK92 cells, but had no significant effect on NK92 cells killing K562 activity. Conclusion IL-21 up-regulates the expression of SENP1 in NK92 cells. Interference with Senp1 can inhibit the proliferation of NK92 cells and promote the apoptosis of cells, meanwhile, it also affects the expression of perforin and its cytotoxic activity on tumor cells.