SDF-1对卵巢癌细胞增殖和侵袭能力作用的相关通路研究

The signal transduction pathway of SDF-1 on proliferation,invasiveness of ovarian carcinoma

目的:探讨PI3K/Akt信号通路抑制剂LY294002、MEK通路抑制剂PD98059对基质衍生因子-1(SDF-1)作用下卵巢癌细胞SKOV3的增殖、侵袭能力的影响,初步探讨SDF-1在卵巢癌中的相关作用通路。方法:细胞免疫化学法检测SDF-1不同作用时间及不同作用浓度对卵巢癌细胞中磷酸化Akt(p-Akt)和ERK(p-ERK)表达的影响。选取对数生长期细胞,分别加SDF-1 100ng/ml、CXCR4中和抗体10μg/ml、CXCR4抑制剂AMD3100 1μg/ml、LY294002 50μmol/L、PD98059 50μmol/L。MTT、Transwell细胞侵袭实验检测细胞的增殖、侵袭能力的变化。结果:随着SDF-1作用时间的延长,p-Akt、p-ERK蛋白表达水平增高;Akt、ERK蛋白的最佳活化时间为30min。作用30min时,随着SDF-1作用浓度的增加,p-ERK1、p-Akt蛋白表达水平增高,差异具有统计学意义(P<0.05)。SDF-1可明显促进SKOV3细胞的增殖、侵袭能力(P<0.05);但加入CXCR4中和抗体、CXCR4抑制剂AMD3100、通路抑制剂PD98059、LY294002后,SKOV3细胞的增殖、侵袭能力无明显变化(P<0.05)。结论:SDF-1对卵巢癌SKOV3细胞的增殖、侵袭能力的增强作用可被PI3K/Akt信号通路抑制剂、MEK通路抑制剂所阻断。SDF-1可能通过Ras/ERK信号转导通路、PI3K/Akt信号通路发挥作用。

Objective:To study the effect of inhibitors of PI3K/Akt signal transduction pathway(LY294002) and MEK signal transduction pathway(PD98059) on proliferation,invasiveness of ovarian carcinoma cell SKOV3 which had been stimulated by SDF-1,and then to explore the the signal transduction pathway of SDF-1.Methods:The expression of p-Akt and PERK were detected by Immunocytochemistry.Add 100ng / ml SDF-1,neutralizing antibody of CXCR4,inhibitors of CXCR4 AMD3100,the inhibitors of signal transduction pathway LY294002,PD98059 to logarithmic growth phase cells,and detected the effect of proliferation,invasiveness of ovarian carcinoma cell among different experimental group by monotetrazolium(MTT) assay and Transwell invasion trial.Results:(1) Longer 100ng/ml SDF-1 acted for,higher the expression of P-ERK and p-Akt protein were.The best acting time was 30 minutes.When concentrations of SDF-1 increased,the expression of P-ERK and p-Akt increased too,and the difference was significant.(2) SDF-1 increased the ability of proliferation,invasiveness of ovarian carcinoma cell,but inhibitors of CXCR4(AMD3100,1μg / ml) 、neutralizing antibody of CXCR4(10μg / ml) 、the inhibitors of RAS / ERK signal transduction pathway(PD98059) and PI3K / Akt signal transduction pathway(LY294002) can blocked the function too(P < 0.05).Conclusion:The effect of SDF-1 on proliferation,invasiveness of ovarian carcinoma cell can be blocked by inhibitors of RAS / ERK and PI3K / Akt signal transduction pathway.SDF-1 may play its functions through Ras / ERK and PI3K / Akt signaling pathway.