C-myc反义寡脱氧核苷酸可诱导类风湿性滑膜细胞凋亡并下调Fas表达。

C-myc antisense oligodeoxynucleotides can induce apoptosis and down-regulate Fas expression in rheumatoid synoviocytes.

作者信息

Hashiramoto A, Sano H, Maekawa T, Kawahito Y, Kimura S, Kusaka Y, Wilder R L, Kato H, Kondo M, Nakajima H

机构信息

Kyoto Prefectural University of Medicine, Japan.

出版信息

Arthritis Rheum. 1999 May;42(5):954-62. doi: 10.1002/1529-0131(199905)42:5<954::AID-ANR14>3.0.CO;2-J.

DOI:10.1002/1529-0131(199905)42:5<954::AID-ANR14>3.0.CO;2-J

PMID:10323451

Abstract

OBJECTIVE

To investigate the role of c-myc in the pathogenesis of rheumatoid arthritis (RA) and the mechanism of synovial apoptosis.

METHODS

Using cultured human synoviocytes from patients with RA and c-myc antisense oligodeoxynucleotides (AS ODN), we examined the inhibition of cell proliferation by the MTT assay and the induction of apoptosis with TUNEL staining and fluorescence microscopy. In addition, the effect of c-myc on down-regulation of Fas expression was analyzed by flow cytometry, cytotoxicity assay, and reverse transcriptase-polymerase chain reaction.

RESULTS

Treatment with c-myc AS ODN induced inhibition of cell proliferation, along with down-regulation of c-Myc protein and c-myc messenger RNA (mRNA) expression. The morphologic changes of synovial cell death were typical of apoptosis. In addition, c-myc AS ODN treatment down-regulated expression of Fas mRNA but not Fas antigen. Analysis of the involvement of the caspase cascade revealed that the cytotoxic activity of c-myc AS ODN was completely blocked by inhibitors of both caspase 1 (YVAD-FMK) and caspase 3 (DEVD-FMK).

CONCLUSION

Our results strongly suggest that c-myc AS ODN might be a useful therapeutic tool in RA and clarify that cell death by c-myc AS ODN is induced through the caspase cascade, similar to Fas-induced apoptosis. In addition, combination therapy with anti-Fas antibody and c-myc AS ODN reduced Fas-dependent cytotoxicity.

摘要

目的

探讨c-myc在类风湿关节炎（RA）发病机制中的作用及滑膜细胞凋亡的机制。

方法

利用培养的RA患者的人滑膜细胞和c-myc反义寡脱氧核苷酸（AS ODN），通过MTT法检测细胞增殖抑制情况，采用TUNEL染色和荧光显微镜观察细胞凋亡诱导情况。此外，通过流式细胞术、细胞毒性测定和逆转录聚合酶链反应分析c-myc对Fas表达下调的影响。

结果

用c-myc AS ODN处理可诱导细胞增殖抑制，同时c-Myc蛋白和c-myc信使核糖核酸（mRNA）表达下调。滑膜细胞死亡的形态学变化是典型的凋亡。此外，c-myc AS ODN处理下调了Fas mRNA的表达，但未下调Fas抗原的表达。对caspase级联反应参与情况的分析表明，c-myc AS ODN的细胞毒性活性被caspase 1（YVAD-FMK）和caspase 3（DEVD-FMK）的抑制剂完全阻断。