›› 2012, Vol. 24 ›› Issue (4): 266-269.doi: 10.3969/j.issn.1004-616x.2012.04.006

• 论著 • 上一篇    下一篇

吲哚胺2，3-双加氧酶上调人肝癌细胞人类白细胞抗原-G的表达

孙建莉;何冬梅;张 敏;任晓静;申慧琴;武希润;王 琦   

  1. ( 1. 山西医科大学研究生院，山西 太原 030001； 2. 山西医科大学附属第二临床医院消化内科，山西 太原 030001 )
  • 收稿日期:2012-03-09 修回日期:2012-03-26 出版日期:2012-07-30 发布日期:2012-07-30
  • 通讯作者: 王 琦

Indoleamine 2, 3-dioxygenase up regulates the expression of human leucocyte antigen-G in hepatocellular carcinoma cells

SUN Jian-li; HE Dong-mei; ZHANG Min; REN Xiao-jing;SHEN Hui-qin;WU Xi-run;WANG Qi   

  1. (1. Graduate School, Shanxi Medical University, Taiyuan 030001; 2. Department of Gastroenterology, Second Affiliated Hospital of Shanxi Medical University, Taiyuan 030001, Shanxi, China)
  • Received:2012-03-09 Revised:2012-03-26 Online:2012-07-30 Published:2012-07-30
  • Contact: WANG Qi

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摘要： 目的： 探讨人类吲哚胺2，3-双加氧酶(indoleamine 2，3-dioxygenase ，IDO)对肝癌细胞人类白细胞抗原-G (human leucocyte antigen-G ，HLA-G)表达的影响。方法：用脂质体lipofectamineTM2000将pcDNA3.1-IDO重组质粒稳定转染入肝癌SMMC-7721细胞作为实验组(pcDNA3.1-IDO质粒转染组)，同时设置空载体对照组(pcDNA3.1转染组)和空白对照组(未转染组)，用RT-PCR和Western blot鉴定实验组和2个对照组细胞IDO mRNA和蛋白的表达，然后分别给予实验组细胞IDO的抑制剂(2.5 mmol/L 1-D-MT)和底物(200 μmol/L L-色氨酸)干预48 h，进一步用实时荧光定量PCR和Western blot检测上述5组细胞中HLA-G mRNA和蛋白的表达水平。结果：RT-PCR和Western blot显示空载体对照组和空白对照组未见IDO表达，而实验组细胞高表达IDO。实时荧光定量PCR和Western blot结果显示，实验组HLA-G mRNA和蛋白表达水平较空载体对照组和空白对照组均明显上调(P均<0.05)，给予1-D-MT和L-色氨酸干预后此效应可逆转。结论：在肝癌SMMC-7721细胞，IDO可以上调HLA-G的表达，色氨酸降解途径参与了此调节过程。

关键词: 人类吲哚胺2, 3-双加氧酶, 人类白细胞抗原-G, SMMC-7721细胞

Abstract: OBJECTIVE: To investigate the effect of indoleamine 2，3-dioxygenase(IDO) on the expression of human leucocyte antigen-G(HLA-G) in hepatocellular carcinoma cells. METHODS： Hepatocellular carcinoma cells SMMC-7721 transfected with pcDNA3.1- IDO recombinant plasmid by lipofectamineTM 2000 reagent as experimental group (pcDNA3.1 -IDO group)， SMMC-7721 transfected with blank plasmid( pcDNA3.1 group) and blank SMMC-7721 (non-transfected group) were used as control groups. The expression of IDO in expreimental group and control groups were tested by RT-PCR and Western blot， the expression of HLA-G mRNA and protein in cells were tested by real time fluorescence quantitative PCR and Western blot. Then pcDNA 3.1-IDO plasmid group cells were treated with IDO competitive inhibitior( 1-methyl-D-trypophan) and its substrate (L-tryptophan) intervention to investigate the effect of drugs on the expression of HLA-G. RESULTS：IDO was highly expressed in pcDNA3.1-IDO plasmid group cells shown by RT-PCR and Western blot. Expression of HLA-G mRNA and protein in pcDNA3.1-IDO plasmid group were higher than the other control groups, the intervention of L-tryptophan and 1-methyl-D-trypophan could reverse this effect. CONCLUSION：IDO could up regulate the expression of HLA-G in SMMC- 7721 cells with tryptophan degradation pathway involved in this regulation.

Key words: indoleamine 2, 3-dioxygenase, human leucocyte antigen-G, SMMC-7721cells

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