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目的:探讨rhPRL对NK细胞系增殖状态及细胞毒活性的调节作用。方法:利用MTT比色法结合细胞计数法观测了rhPRL对三种NK细胞系(NKL、NK-92、YT)增殖活性和细胞毒活性的影响;RT-PCR法、直接免疫荧光法和FACS技术检测了PRL-R和CD25在不同细胞系的表达状况以及rhPRL对CD25表达强度的影响。结果:MTT结果表明,在低剂量IL-2存在的条件下,rhPRL浓度为12和100ng/ml时对NK-92和NKL表现出最佳刺激效应,该反应呈双峰型曲线。相反,YT细胞的增殖状态和细胞毒效应均无明显改变。当有足量IL-2(100 U/ml)存在时,rhPRL对三种 NK细胞系均无明显刺激作用。RT-PCR和直接免疫荧光结果显示,三种细胞系均有PBL-R表达,其表达量以NKL细胞最高,NK-92和YT荧光强度较弱。FACS结果表明CD25在NKL表达量最高约90%,NK-92次之约为50%,YT最低为10%,rhPRL刺激后其表达明显上调。结论:rhPRL能促进NK细胞系(NK-92和NKL)的增殖活性和杀伤活性,该效应可能部分是通过IL-2受体表达并参与其下游信号呈递通路实现的。
Objective: To investigate the regulatory effect of rhPRL on proliferation and cytotoxicity of NK cell line. Methods: The effects of rhPRL on the proliferation and cytotoxicity of three NK cell lines (NKL, NK-92, YT) were observed by MTT assay and cell counting method. RT-PCR, direct immunofluorescence and FACS The expression of PRL-R and CD25 in different cell lines and the effect of rhPRL on the expression of CD25 were examined. Results: MTT results showed that rhPRL at 12 and 100 ng / ml showed the best stimulation effect on NK-92 and NKL in the presence of low dose of IL-2, and the reaction showed a bimodal curve. In contrast, there was no significant change in the proliferation status and cytotoxicity of YT cells. When sufficient IL-2 (100 U / ml) was present, rhPRL had no significant stimulatory effect on all three NK cell lines. The results of RT-PCR and direct immunofluorescence showed that PBL-R was expressed in all the three cell lines, with the highest expression in NKL cells and weak fluorescence intensity of NK-92 and YT. FACS results showed that the expression level of CD25 in NKL was about 90%, that in NK-92 was about 50%, and that in YT was 10%. After rhPRL stimulation, the expression of CD25 was up-regulated. CONCLUSION: rhPRL can promote the proliferation and cytotoxicity of NK cell lines (NK-92 and NKL). This effect may be partially mediated by the expression of IL-2 receptor and its involvement in downstream signaling pathways.