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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 investigate the effect of PI3K / Akt signaling pathway inhibitor LY294002 and MEK pathway inhibitor PD98059 on proliferation and invasion of ovarian cancer cell line SKOV3 under the action of SDF-1. To investigate the effect of SDF-1 on ovarian cancer In the relevant role of access. Methods: The effects of different time and concentration of SDF-1 on the expression of phosphorylated Akt (p-Akt) and ERK (p-ERK) in ovarian cancer cells were detected by immunocytochemistry. The cells in the logarithmic growth phase were selected and treated with 100 ng / ml SDF-1, 10 μg / ml CXCR4 neutralizing antibody, 1 μg / ml CXCR4 inhibitor, 50 μmol / L LY294002 and 50 μmol / L PD98059 respectively. MTT, Transwell cell invasion assay to detect cell proliferation, invasive ability changes. Results: The expression of p-Akt and p-ERK increased with the prolongation of SDF-1 treatment. The optimal activation time of Akt and ERK protein was 30 min. With the increase of SDF-1 concentration, the expression of p-ERK1 and p-Akt protein increased at 30min (P <0.05). SDF-1 could significantly promote the proliferation and invasion ability of SKOV3 cells (P <0.05). However, the proliferation and invasion ability of SKOV3 cells were not significantly changed after adding CXCR4 neutralizing antibody, CXCR4 inhibitor AMD3100, PD98059 and LY294002 P <0.05). Conclusion: The enhanced effect of SDF-1 on proliferation and invasion of ovarian cancer SKOV3 cells can be blocked by inhibitors of PI3K / Akt signaling pathway and MEK pathway inhibitors. SDF-1 may play a role through the Ras / ERK signal transduction pathway and the PI3K / Akt signaling pathway.