[1]张维,李宝金,伊远学,等.小干扰RNA特异性抑制淋巴细胞T-bet表达的研究[J].第三军医大学学报,2009,31(13):1291-1294.
 ZHANG Wei,LI Bao-jin,YI Yuan-xue,et al.Study on the expression of T-bet on human lymphocytes inhibited by specific siRNA[J].J Third Mil Med Univ,2009,31(13):1291-1294.
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小干扰RNA特异性抑制淋巴细胞T-bet表达的研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第13期
页码:
1291-1294
栏目:
论著
出版日期:
2009-07-15

文章信息/Info

Title:
Study on the expression of T-bet on human lymphocytes inhibited by specific siRNA
作者:
张维李宝金伊远学吴立旋刘晓平陶剑平
北京大学深圳医院肝胆腔镜外科;中山大学中山医学院微生物学教研室
Author(s):
ZHANG Wei LI Bao-jin YI Yuan-xue WU Li-xuan LIU Xiao-ping TAO Jian-ping
Department of Hepatobiliary Laparoscopy Surgery, Shenzhen Hospital, Peking University, Shenzhen 518036, Guangdong Province; Department of Microbiology, Zhongshan Medical College, Sun Yet-sen University, Guangzhou 510080, China
关键词:
RNA干扰小干扰RNA淋巴细胞T-bet
Keywords:
RNA interference small interfering RNA lymphocyte T-bet
分类号:
R392-33; R392.12; R392.4
文献标志码:
A
摘要:
目的   探讨体外转录法合成的小干扰RNA(small interfering RNA,siRNA)对人淋巴细胞T-bet基因表达及功能的影响。   方法   设计并合成4条siRNA(siRNA-1、siRNA-2、siRNA-3及siRNA-co),脂质体法转染淋巴细胞。转染后48 h 收集转染后细胞,采用RT-PCR方法检测细胞T-bet mRNA的抑制率;酶联免疫吸附试验(ELISA)方法检测细胞上清液中IL-4和IFN-γ水平;将淋巴细胞与人脐静脉血管内皮细胞(ECV304)共培养检测淋巴细胞对ECV304增殖的抑制作用。   结果   转染后48h半定量RT-PCR的检测显示siRNA-1、siRNA-2和siRNA-3转染后的淋巴细胞T-bet表达抑制率分别为(72.50±1.01)%、(5.40±0.63)%和(30.90±0.90)%,以siRNA-1转染后的淋巴细胞T-bet表达抑制作用最强(P<0.05),其细胞上清液中IL-4水平最高,而IFN-γ的水平最低(P<0.05),siRNA-1转染淋巴细胞对人脐静脉血管内皮细胞(ECV304)增殖的抑制作用低于siRNA-2及siRNA-3。   结论   siRNA可特异性抑制淋巴细胞T-bet基因的表达,从而为进一步研究siRNA在移植免疫耐受中应用提供了理论和实验基础。
Abstract:
Objective   To investigate the expression of T-bet on human lymphocytes inhibited by siRNA.     Methods   Four different siRNAs (siRNA-1, siRNA-2, siRNA-3, and siRNA-co) were designed and synthysized and transfected into freshly isolated human lymphocytes with cationic liposome. The changes of T-bet mRNA levels were determined by semi-quantitative RT-PCR at 48 h after transfection. IL-4 and IFN-γ cytokine levels were assayed by ELISA. The proliferation of ECV304 co-cultured with T lymphocyte by cck-8 assay was determined.     Results   Different siRNA showed different reduction in T-bet expression. At 48 h after transfection, the degrees of reduction with siRNA-1, siRNA-2, and siRNA-3 were (72.50±1.01) %, (5.40±0.63)%, and (30.90±0.90)%, respectively as compared with those of the control (P<0.05). siRNA-1 was the most efficient. The results of semi-quantitative RT-PCR assay indicated T-bet mRNA levels were inhibited after transfection. IL-4 cytokine level in the transfection group was higher than that in the control group, but IFN-γ cytokine level in the transfection group was lower than that in the control group. The proliferation level of ECV304 co-cultured with T lymphocyte transfected with siRNA-1 group was higher than that of siRNA-2 group and siRNA-3 group.     Conclusion   Three different siRNAs could reduce the expression of T-bet and the T-bet mRNA level. siRNA may be useful for further study on graft-versus-host disease after allogeneic transplantation.

参考文献/References:

张维, 李宝金, 伊远学, 等.小干扰RNA特异性抑制淋巴细胞T-bet表达的研究[J]. 第三军医大学学报,2009,31(13):1291-1294.

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更新日期/Last Update: 2009-06-17