酸性成纤维细胞生长因子在卵巢癌中的表达及其信号传导途径

目的　探讨酸性成纤维细胞生长因子 (aFGF)在卵巢癌发生发展中的作用及其信号传导机制。方法　应用逆转录 聚合酶链反应技术 (RT PCR)和Western印迹方法对 40例上皮性卵巢癌组织aFGF及其受体FGFR1的表达进行了分析 ;并以不同浓度的aFGF和酪氨酸蛋白激酶 (TPK)抑制剂 genistein诱导卵巢癌细胞株CAOV3细胞 ,利用 [γ 32 P]ATP掺入外源性底物的方法 ,液体闪烁测定蛋白激酶C(PKC)及细胞外信号调节激酶 (ERK)的活性。结果　aFGFmRNA和FGFR1mRNA在上皮性卵巢癌组织中的半定量检测结果分别为 0 981± 0 1 30和 1 0 4 7± 0 1 4 8,与正常卵巢、卵巢瘤样病变及卵巢良性肿瘤相比 ,差异有显著意义 (P <0 0 5) ,且在Ⅲ～Ⅳ期卵巢癌中的表达水平明显高于Ⅰ～Ⅱ期 (P <0 0 5) ;Western印迹检测见上皮性卵巢癌组织中aFGF和FGFR1条带均深于正常卵巢组织 ;随着aFGF浓度的增加 ,CAOV3细胞PKC及ERK活性随之升高 ,与aFGF浓度呈剂量依赖效应 ;Genistein抑制细胞内PKC及ERK活性 ,与 genistein浓度亦呈剂量依赖效应。 结论　aFGF与上皮性卵巢癌的发生、发展、浸润呈正相关 ,其受体具有TPK活性 ,TPK激活后可进一步激活PKC和ERK ,进一步证明PKC及ERK确是TPK的下游信号分子

Expression of aFGF in ovarian epithelial cancer and its signal transduction pathway

OBJECTIVE: To explore the expression of acidic fibroblast growth factor (aFGF) and its receptor FGFR1 in ovarian epithelial cancer and observe the effects of aFGF and TPK inhibitor Genistein on intracellular PKC and ERK activity in ovarian epithelial cancer cells line CAOV3. METHODS: The expression levels of aFGF and FGFR1 were evaluated by RT-PCR and western blot in 40 cases of ovarian epithelial cancer. The activity of PKC and ERK in cells induced by different concentration of aFGF and Genistein were detected by incorporation of [gamma-(32)P]-ATP into exogenous substrate. RESULTS: The expression levels of aFGFmRNA and FGFR1mRNA in the ovarian epithelial cancer were 0.981 +/- 0.130 and 1.047 +/- 0.148, respectively. Compared with normal ovary, ovarian tumor like condition and benign ovary tumors, the difference was significant (P < 0.05). The expression levels in stage III - IV were significantly higher than those in stage I - II (P < 0.05). There were overexpression of aFGF and FGFR1 in the ovarian epithelial cancer in western blot, too. The intracellular PKC and ERK activity increased with aFGF in a dose dependent manner, Genistein suppressed the intracellular PKC and ERK activity also in a dose dependent manner. CONCLUSION: aFGF may play an important role in carcinogenesis, development and invasion of ovarian epithelial cancer. Its receptor in human ovarian cancer cell line CAOV3 possessed TPK activity. These tyrosine-specific protein phosphorylation may initiate a cascade of biochemical events, which may increase the intracellular PKC and ERK activity. PKC and ERK locate downstream of TPK in CAOV3 cell line.