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根据已知的PTKs的氨基酸序列保守区设计简并引物 ,采用 3′快速末端扩增法 (3′RACE)扩增了滑膜细胞中PTKscDNAs的 3′末端 ,然后通过RNA点杂交方法分别观察了这些PTKs在类风湿性关节炎 (RA)和骨性关节炎 (OA)病人滑膜细胞中的表达水平。结果从RAFLS细胞中共克隆到 6种已知PTKscDNA片段 ,分别为血小板衍生生长因子受体A (PDGFRA)、胰岛素样生长因子 1受体 (IGF 1R)、含盘状 (discoidin)结构域的受体型酪氨酸激酶 (DDR2 )、Lyn、Janus激酶 1(JAK1)和TYK2。RNA点杂交结果显示 ,在 4例RA病人和 2例OA病人的滑膜细胞中都表现为 :PDGFRA、IGF 1R和DDR2在RA滑膜细胞中的表达水平高于OA滑膜细胞 ,其它几种激酶在两种细胞中的表达水平无明显差别 ,说明RA滑膜细胞中至少表达PDGFR、IGF 1R、Lyn、DDR2、JAK1和TYK2等 6种PTKs,其中PDGFRA、IGF 1R和DDR2可能与RA滑膜细胞的过度增殖和对软骨的侵蚀性相关。
Degenerate primers were designed according to the conserved regions of the known amino acid sequences of PTKs. The 3 ’end of PTKscDNAs in synovial cells was amplified by 3’ rapid amplification (3’RACE) and then observed by RNA dot blot The expression levels of these PTKs in synovial cells of patients with rheumatoid arthritis (RA) and osteoarthritis (OA). Results Six kinds of known PTK cDNA fragments were co-cloned from RAFLS cells. They were PDGFRA, IGF 1R, and discoidin domain Tyrosine kinase (DDR2), Lyn, Janus kinase 1 (JAK1), and TYK2. RNA dot blot results showed that the expression of PDGFRA, IGF 1R and DDR2 in RA synovial cells was higher in synovial cells of 4 RA patients and 2 OA patients than that of OA synovial cells. The other several The expression level of kinases in two kinds of cells showed no significant difference, indicating that at least six kinds of PTKs such as PDGFR, IGF1R, Lyn, DDR2, JAK1 and TYK2 were expressed in RA synovial cells. PDGFRA, IGF1R and DDR2 may be associated with RA synovial membrane Cell hyperproliferation and cartilage erosion related.