黏着斑激酶通过细胞周期蛋白依赖性激酶2、c-junN端激酶促进人肺动脉平滑肌细胞增殖  

CDK and JNK mediate FAK to enhance proliferation in human pulmonary artery smooth cells

作　　者：林春龙[1] 

机构地区：[1]湖南师范大学附属岳阳医院,岳阳市立二医院呼吸内科,湖南岳阳414000

出　　处：《中国病理生理杂志》2008年第2期303-306,共4页Chinese Journal of Pathophysiology

摘　　要：目的:探讨黏着斑激酶(FAK)促进人肺动脉平滑肌细胞(HPASMCs)增殖的机制。方法:取肺癌部分切除手术病人远离肺癌的周围正常组织,酶消化法培养细胞。在正义寡核苷酸(FAK SODNs)、反义寡核苷酸(FAK ASODNs)、随机寡核苷酸(FAK MS)转染后,用免疫印迹方法测定了FAK、c-junN端激酶(JNK)、细胞周期蛋白依赖性激酶2(CDK2)的表达;用流式细胞仪测定了细胞周期、细胞凋亡;用免疫组织化学法测定胞浆FAK的表达。结果:免疫印迹结果表明,与FAK MS组比较,应用FAK ASODNs转染后HPASMCs FAK、JNK、CDK2蛋白质的表达明显降低(P<0.01、P<0.05、P<0.01);而用正义寡核苷酸转染后则明显增加(P<0.01、P<0.05、P<0.05)。流式细胞仪测定结果示:与FAK MS转染组比较:FAK SODNs转染后,HPASMCs中G1相的比例显著减少,S相显著增加(P<0.01),细胞凋亡减少(P<0.01);而FAK ASODNs转染后结果则相反。免疫组织化学结果显示:FAK SODNs转染后HPASMCs胞浆染色增强,FAK ASODNs转染后胞浆浅染,提示FAK表达减少。结论:FAK可能经由JNK、CDK2信号通路促使细胞增殖。AIM： To find out the mechanism of focal adhesion kinase （FAK） facilitating human pulmonary artery smooth muscle cells （HPASMCs） proliferation. METHODS ： HPASMCs were isolated from normal part of lungs of two carcinoma patients who undergone lung partial resection. Cultured HPASMCs stimulated by fibronection（40 mg/L） were passively transfected with ODNs, sense focal adhesion kinase （FAK）, mismatch sense and antisense -FAK, respectively. Expression of FAK, Jun NH2 -terminal kinase （JNK） and cyclin -dependent kinase2 （CDK2） proteins were detected by immunoprecipitation and Western blotting. Cell cycle and cell apoptosis were analyzed by flow cytometry. In addition, cytoplasma FAK expression was detected by immunohistochemistry staining. RESULTS ： The protein expressions of FAK, JNK and CDK2 in HPASMCs decreased in FAK ASODNs group and increased in FAK SODNs group. Meanwhile, the proportion of cells at G1 phase decreased significantly in FAK SODNs group, while the cells at S phase increased significantly. In contrast, the proportion of cells at G1 phase was increased significantly in FAK ASODNs group. The level of cell apoptosis in FAK ASODNs group was higher. FAK expression in FAK SODNs group was strongly stained by immunocytochemistry, whereas that in FAK ASODNs group was weakly stained. CONCLUSION： The results suggest that FAK via JNK, CDK2 signaling pathway enhances HPASMCs proliferation.

关 键 词：黏着斑激酶 人肺动脉平滑肌细胞 C-JUN N端激酶 细胞周期蛋白依赖激酶类 

分 类 号：R363[医药卫生—病理学]