【摘要】 目的:探讨血小板活化因子(PAF)对卵巢癌细胞增殖的影响及其相关机制。方法:用不同浓度PAF及阻断剂作用于浆液性卵巢癌细胞株OVCA 429及黏液性卵巢癌细胞株RMUG-L,MTT法测定细胞增殖情况,Western blot法测定EGFR、Src、FAK、Paxillin及相应磷酸化蛋白表达。免疫共沉淀(Co-IP)测定PAFR与EGFR的相互作用。结果:PAF促进PAFR阳性卵巢癌细胞株OVCA 429细胞增殖,可被各种PAFR阻断剂阻断,诱导PAFR阳性细胞EGFR磷酸化,而对PAFR阴性细胞株无这一作用。PAF作用后未能检测到PAFR与EGFR直接相互作用。PAF可诱导下游Src、FAK及Paxillin蛋白的磷酸化。结论:PAF促进PAFR阳性卵巢癌细胞增殖是受体依赖的。PAFR激活后可同时激活EGFR,导致关键蛋白Src磷酸化,后者进而活化FAK与Paxillin,进一步激活下游蛋白,从而促进细胞增殖。更多还原

【Abstract】 Objective:To explore the effects and mechanisms of PAF on the proliferation of ovarian cancer cells.Methods:The ovarian cancer cell lines were treated by PAF alone or in combination with inhibitors.The proliferation of cells was determined by MTT assay.The cells were treated by PAF at different time and the expressions of EGFR and p-EGFR was determined by Western blot.Co-immunoprecipitation(Co-IP) was used to determine whether there was a direct interaction between PAFR and EGFR.The expressions of Src,FAK,Paxillin and their corresponding phosphorylated proteins was tested.Results:The proliferation of PAFR positive cells was significantly increased when treated by PAF.The effect of PAF promoting cell proliferation could be blocked by GB and PAFR Ab.The EGFR was phosphorylated by PAF in PAFR positive cells,but not in PAFR negative cells.There was no direct interaction between PAFR and EGFR when stimulated by PAF.The phosphorylation levels of Src,FAK and Paxillin were elevated in PAF treatment group.Conclusion:The promotion of proliferation of PAFR positive ovarian cancer cell by PAF was receptor-dependent.The activated PAFR led to the phosphorylation of EGFR and Src,which in turn activated FAK and paxillin.The signal was transferred to the nucleus and thus promoting cell proliferation.更多还原