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目的探讨K562细胞中survivin基因的表达对淋巴细胞增殖和功能的影响。方法将融合重组质粒pEGFP-C1-survivin和靶向survivin基因的RNAi的重组质粒pTZU6+1-survivin分别转染K562细胞,得到K562/survivin+和K562/survivin-2种细胞,用G418筛选K562/survivin+细胞,用半定量逆转录-聚合酶链反应(RT-PCR)方法及免疫细胞化学法检测K562/survivin+和K562/survivin-2种细胞中survivinmRNA及蛋白水平的表达,并与K562细胞作对照。将这3种细胞与健康人外周血单个核细胞(PBMC)混合培养(MLTC),检测混合培养体系中淋巴细胞增殖能力、NK细胞活性及培养上清中IFN-γ水平。结果K562/survivin+组的淋巴细胞增殖指数明显低于其余两组。survivin基因的表达水平与NK细胞活性呈负相关。混合培养上清中分泌的IFN-γ,K562/survivin+组是K562组的1/4.5,是K562/survivin-组的1/8。结论高表达的survivin可以抑制淋巴细胞的增殖、NK细胞活性及IFN-γ的分泌。
Objective To investigate the effect of survivin gene expression on the proliferation and function of lymphocytes in K562 cells. Methods K562 / K562 / survivin-K562 / survivin-2 cells were transfected with pTZU6 + 1-survivin, and the recombinant plasmid pEGFP-C1-survivin and survivin gene were transfected into K562 cells. The expression of survivin mRNA and protein in K562 / survivin + and K562 / survivin-2 cells was detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry. The three kinds of cells and healthy human peripheral blood mononuclear cells (PBMC) were mixed culture (MLTC), detection of mixed culture system in lymphocyte proliferation capacity, NK cell activity and the level of IFN-γ in the culture supernatant. Results The lymphocyte proliferation index of K562 / survivin + group was significantly lower than that of the other two groups. The expression of survivin gene was negatively correlated with the activity of NK cells. IFN-γ secreted in the mixed culture supernatant was 1 / 4.5 in the K562 group and 1/8 of the K562 / survivin-treated group in the K562 / survivin + group. Conclusion High expression of survivin can inhibit lymphocyte proliferation, NK cell activity and IFN-γ secretion.