Miyazawa K, Mori A, Okudaira H
Central Research Laboratories, Kissei Pharmaceutical Co., Ltd., Nagano, 399-8304, Japan.
J Biochem. 1998 Dec 1;124(6):1153-62. doi: 10.1093/oxfordjournals.jbchem.a022233.
A novel immortalized rheumatoid fibroblast-like synoviocyte (FLS) line, MH7A, was established by stably transfecting FLS cells with SV40 T antigen gene. MH7A cells expressed SV40-specific small t and large T antigens as well as an elevated level of p53 protein. They have already reached over 150 population doublings through culture crisis, and have been growing rapidly compared with the parental FLSs. Constitutive activation of p42/p44 mitogen-activated protein (MAP) kinase was detected in MH7A cells. Serum requirements for the growth of MH7A were markedly decreased compared with those for the parental FLSs. MH7A cells were stained positively for interleukin (IL)-1R, intercellular adhesion molecule-1 (ICAM-1), CD16, CD40, CD80, and CD95. IL-1beta enhanced the production of IL-6 and stromelysin-1, and the surface expression of ICAM-1, in a manner similar to that in the parental FLSs. SB203580, a specific inhibitor of p38 MAP kinase, significantly inhibited IL-1beta-induced IL-6 and stromelysin-1 production by both parental FLSs and MH7A cells; although PD098059, an inhibitor of the p42/p44 MAP kinase pathway, did not affect it. Our results clearly indicate the usefulness of MH7A cells for investigating the regulation of rheumatoid FLSs and the IL-1 signal transduction pathway to develop future RA therapy.
通过用SV40 T抗原基因稳定转染成纤维样滑膜细胞（FLS），建立了一种新的永生化类风湿性FLS细胞系MH7A。MH7A细胞表达SV40特异性小t和大T抗原以及高水平的p53蛋白。它们已经通过培养危机达到了超过150次群体倍增，并且与亲代FLS相比生长迅速。在MH7A细胞中检测到p42/p44丝裂原活化蛋白（MAP）激酶的组成性激活。与亲代FLS相比，MH7A生长对血清的需求明显降低。MH7A细胞对白介素（IL）-1R、细胞间粘附分子-1（ICAM-1）、CD16、CD40、CD80和CD95染色呈阳性。IL-1β以与亲代FLS相似的方式增强了IL-6和基质溶解素-1的产生以及ICAM-1的表面表达。p38 MAP激酶的特异性抑制剂SB203580显著抑制了亲代FLS和MH7A细胞中IL-1β诱导的IL-6和基质溶解素-1产生；而p42/p44 MAP激酶途径抑制剂PD098059对此没有影响。我们的结果清楚地表明MH7A细胞对于研究类风湿性FLS调控和IL-1信号转导途径以开发未来类风湿性关节炎治疗方法具有实用性。
