CXCR4及其配体SDF-1α在卵巢癌SKOV3细胞黏附和侵袭中的作用

The role of CXCR4 and its ligand SDF-1α in the process of adhesion and invasion of ovarian cancer

目的:探讨趋化因子受体4(chemokine receptor 4,CXCR4)与其配体基质细胞衍生因子-1α(stromal derived factor-1α,SDF-1α)通过激活丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路在卵巢癌黏附和侵袭中的作用及其机制。方法:采用激光共聚焦显微镜观察SDF-1α处理前、后卵巢癌SKOV3细胞中钙离子的波动;Western印迹法检测SDF-1α对SKOV3细胞中细胞外调节信号激酶(extracellular signal-regulated kinase,ERK)-1和-2磷酸化的影响;通过黏附实验、明胶酶谱法检测SDF-1α/CXCR4对卵巢癌细胞黏附能力以及基质金属蛋白酶(matrix metalloproteinase,MMP)-2和MMP-9活性的调控。结果:SDF-1α诱导卵巢癌细胞内钙的迅速动员及ERK1和ERK2的快速磷酸化;增加卵巢癌细胞黏附于纤维连接蛋白(fibronectin,FN)和Ⅳ型胶原的能力;加入ERK1/2信号通路的抑制剂PD98059后,可降低SDF-1α促进卵巢癌细胞黏附于细胞外基质(extracellular matrix,ECM)的能力;同时SDF-1α可促进卵巢癌细胞分泌活性MMP-2和MMP-9。结论:SDF-1α/CXCR4通过激活MAPK信号通路调控卵巢癌细胞的黏附能力,促进活性MMP-2和MMP-9的分泌,进而参与卵巢癌的侵袭和转移。

Objective：To explore the role of chemokine receptor 4（CXCR4） and its ligand stromal derived factor-1α（SDF-1α） in the processes of adhesion and invasion of ovarian cancer cells through activating the mitogen-activated protein kinases（MAPK） signaling pathway and its possible mechanism.Methods：Intracellular calcium mobilization in SKOV3 cells was detected with a laser scanning confocal fluorescence microscopy before and after SDF-1α treatment.Western blotting was used to detect the phosphorylation of extracellular signal-regulated kinase（ERK1/2） in SKOV3 cells after exposure to SDF-1α.Adhesion capability and matrix metalloproteinase（MMP） activity of ovarian cancer cells after exposure to SDF-1α were mesured by adhesion assay and gelatin zymography,respectively.Results：SDF-1α induced rapid intracellular calcium mobilization in a metastatic ovarian cancer cell line SKOV3,as well as rapid phosphorylation of ERK-1/2.Adhesion capability of ovarian cancer cells to fibronectin（FN） and collagen Ⅳ（COL） was increased by SDF-1α treatment,while the addition of PD98059,an inhibitor of ERK-1/2 signaling,reduced the effects of SDF-1α.SDF-1α also increased the activities of MMP-2 and MMP-9 secreted by SKOV3 cells.Conclusion：SDF-1α/CXCR4 regulates adhesion ability of ovarian cancer cells by activating MAPK signaling pathway and stimulates secretion of MMP-2 and MMP-9,thereby participates the processes of invasion and metastasis of ovarian cancer.