大豆异黄酮对人乳腺癌MCF-7细胞侵袭转移能力的影响及其与PPAR γ的关系研究

EFFECTS OF SOY ISOFLAVONES ON THE INVASION AND METASTASIS OF HUMAN BREAST CANCER MCF-7 CELLS AND ITS CORRELATION WITH PPAR γ

目的研究两种大豆异黄酮成分染料木黄酮(genistein,GEN)和大豆苷元(daidzein,DAI)对人乳腺癌MCF-7细胞体外侵袭转移能力的影响,并初步探讨过氧化物酶体增殖物激活受体γ(PPAR γ)信号途径在其中的作用。方法GEN、DAI单独或联合PPAR γ选择性抑制剂GW9662处理人乳腺癌MCF-7细胞,细胞计数试剂盒(cell counting kit-8,CCK-8)法检测其粘附能力的变化;Millcell膜侵袭系统观察其侵袭能力的改变;免疫细胞化学染色分析局部粘着斑激酶(FAK)、尿激酶型纤维酶原激活物(uPA)蛋白表达变化;激光共聚焦显微镜观察细胞骨架的改变。结果 8×10-5mol/L的GEN、DAI单独作用48h后,MCF-7细胞的粘附率和侵袭细胞数均显著降低;FAK和uPA的蛋白表达水平明显下降;细胞骨架受损。1×10-5mol/L的GW9662与GEN、DAI联合作用时,GEN、DAI对MCF-7细胞的上述作用明显削弱。结论大豆异黄酮具有抑制MCF-7细胞侵袭转移的作用,这种抑制作用与其激活PPAR γ信号途径,降低FAK、uPA表达,影响细胞骨架有关。

Objective To investigate the effects of two major soy isoflavones genistein（GEN）,daizein（DAI） on the invasion and metastasis of human breast cells MCF-7 in vitro,and its correlation with peroxisome proliferators-activated receptor γ（PPAR γ）signal pathway.Method MCF-7 cells were treated with GEN and DAI alone or in combination with GW 9662,a selective inhibitor of PPAR γ.The adhesion ability was determined by cell counting kit-8（CCK-8） assay.Invasion was obsereved in the Millcell Chamber membrane invasion culture system.The expression of focal adhesion kinase（FAK） and urokinase plasminogen（uPA） was detected by immunocytochemical staining.The change of cytoskeletons was observed with laser scanning confocal microscopy.Results The adhension rate,the number of invading MCF-7 cells,and the expression of FAK and uPA were significantly decreased,and the cytoskeletal structure of MCF-7 cells was damaged after treatment with GEN or DAI alone.However,the above effects induced by GEN or DAI were significantly attenuated by GW9662.Conclusion Soy isoflavones inhibit the invasion and metastasis of breast cancer MCF-7 cells in vitro by activating PPAR γ signal pathway,down-regulating FAK and uPA expression,and affecting the cytoskeletal structure.