【作者】 沈晓燕；

【导师】 王泽华；

【作者基本信息】 华中科技大学 ， 妇产科学， 2007， 硕士

【摘要】 目的探讨SDF-1/CXCR4轴通过激活ERK1/2信号通路对卵巢癌转移的影响。方法采用激光共聚焦显微镜检测加SDF-1α前后细胞内钙离子的波动,Western blot法分析SDF-1α对SKOV3细胞ERK1/2磷酸化的影响,分别通过黏附实验和明胶酶谱法检测SDF-1/CXCR4对卵巢癌细胞SKOV3黏附能力和基质金属蛋白酶(MMP)的分泌的影响。结果SDF-1α诱导了卵巢癌细胞胞内钙的迅速动员,钙波测定表明加入SDF-1α20秒钙离子开始升高,200秒达高峰;SDF-1α也诱导了ERK1/2的快速磷酸化,western blot实验表明加入SDF-1α5min ERK1/2开始出现磷酸化,5min至30min磷酸化程度逐渐增强,表明SDF-1α与卵巢癌细胞表面CXCR4的相互作用可以活化对肿瘤细胞增殖和转移有重要作用的ERK1/2信号通路,SDF-1α增加了卵巢癌细胞黏附于纤维连接蛋白(FN)和Ⅳ型胶原(COL)的能力,加入ERK1/2信号通路的抑制剂PD98059后,降低了SDF-1α促进卵巢癌细胞黏附于细胞外基质蛋白(ECM)的作用, SDF-1α也促进了卵巢癌细胞分泌活性的MMP-2和MMP-9。结论卵巢癌侵袭和转移在一定程度上依赖于SDF-1/CXCR4的相互作用,SDF-1增加卵巢癌细胞株SKOV3的侵袭力和转移力,这些作用至少部分是通过激活ERK1/2信号通路使细胞的黏附能力增加和促进MMP-2、MMP-9的分泌实现的。提示通过对MAPK信号通路的降调节可能发挥抑制卵巢癌的转移的作用。更多还原

【Abstract】 Objective: To explore the role of SDF-1/CXCR4 in mediating the Metastasis of ovarian cancer cell through activation of ERK1/2 signal pathway.Methods:Intracellular calcium mobilization was detected with a Laser Scanning Confocal Fluorescence Microscopy. Western blotting was used to detect the phospharylation of 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-1αwere mesured by adhesion assay and gelatin zymography.Results:SDF-1αinduced rapid intracellular calcium mobilization in a metastatic ovarian cancer cell line SKOV3. Calcium mobilization assay show that Calcium flux rapidly increased after 20 second of exposure to SDF-1αand the peak time of Calcium flux was 200 second. SDF-1αalso induced rapid phosphorylation of ERK-1/2. Western blotting showed that ERK-1/2 was phosphorylated in SKOV3 cells after 5 minutes of exposure to SDF-1α. There was gradually increase in the ratio of phospho-ERK at 0 minute to at 30 minutes.SDF-1/CXCR4 interaction can activate ERK signal pathway that plays an important role in regulating cell proliferation and migration. Ovarian cancer cell adhesion to fibronectin and collagen IV, increased after SDF-1αtreatment, while the addition of an inhibitor of ERK-1/2 signaling, PD98059, reduced the effects of SDF-1αSDF-1αalso increased the active MMP-2 and MMP-9 secreted.Conclusion:The SDF-1/CXCR4 axis plays a critical role in the migration and metastasis of human ovarian cancer by regulating the adhesion capability and MMP-2 and MMP-9 activity through ERK1/2 signal pathway.Together, these studies reveal important cellular and molecular mechanisms of SDF-1/CXCR4-mediated ovarian cancer migration and invasion and investigate the ways to block this process by inhabiting the MAPK signaling transduction pathway.更多还原