[1]罗颖,温贤浩,徐酉华.K562细胞survivin基因表达对淋巴细胞增殖和功能的影响[J].第三军医大学学报,2008,30(23):2215-2218.
 LUO Ying,WEN Xian-hao,XU You-hua.Effect of enhanced survivin expression on lymphocyte proliferation and function in K562 cells[J].J Third Mil Med Univ,2008,30(23):2215-2218.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第23期
页码:
2215-2218
栏目:
论著
出版日期:
2008-12-15

文章信息/Info

Title:
Effect of enhanced survivin expression on lymphocyte proliferation and function in K562 cells
作者:
罗颖 温贤浩 徐酉华
重庆医科大学附属儿童医院血液科
Author(s):
LUO Ying WEN Xian-hao XU You-hua
Department of Hematology, Children’s Hospital, Chongqing Medical University, Chongqing 400014, China
关键词:
survivin淋巴细胞增殖K562细胞RNA干扰
Keywords:
survivin lymphocyte proliferation K562 cells RNA interference
分类号:
R331.144; R73-362; R733.7
文献标志码:
A
摘要:
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survivin mRNA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。
Abstract:
Objective    To study the effect of survivin gene expression on the lymphocyte’s proliferation and function in cultured K562 cells.     Methods    The constructed recombinant vector pEGFP-C1-survivin and the plasmid pTZU6+1-survivin encoding short hairpin RNA of survivin were transfected into K562 cells respectively to generate K562/survivin+ cells and K562/survivin- cells. K562/survivin+ cells were selected by G418. The survivin mRNA and protein levels in the 3 kinds of cells (K562/survivin+, K562 and K562/survivin-) were detected by semi-quantitative RT-PCR and immunohistochemistry methods. The peripheral blood mononuclear cells (PBMC) from healthy subjects were co-cultured with these 3 cells respectively in mixed lymphocyte-tumor cell culture (MLTC). Lymphocyte proliferation in the supernatant was evaluated by MTT assay. Nature killer activity was detected by flow cytometry and IFN-γ level was measured by ELISA assay.     Results    Proliferation index in K562/survivin+ group was much lower than that in the other 2 groups (P<0.05). Survivin expression level was of negative correlation with NK activity. IFN-γ content in the MLTC of K562/survivin+ group was lower than that in K562 group (1/4.5) and K562/survivin- group (1/8).     Conclusion    Enhanced expression of survivin may inhibit the proliferation of lymphocyte, activity of nature killer and the secretion of IFN-γ.

参考文献/References:

罗颖, 温贤浩, 徐酉华. K562细胞survivin基因表达对淋巴细胞增殖和功能的影响[J].  第三军医大学学报, 2008, 30(23):2215-2218.

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更新日期/Last Update: 2008-11-12