去甲斑蝥素抑制cortactin、MMP-9表达及降低前列腺癌PC-3细胞侵袭力的研究

Norcantharidin Inhibits the Expression of Cortactin and MMP-9 and Decreases the Invasion of PC-3 Cells

目的观察去甲斑蝥素(NCTD)对前列腺癌PC-3细胞迁移侵袭能力以及cortactin、MMP-9mRNA及蛋白表达的影响。方法取对数生长期的PC-3细胞,分为模型组(无处理)、阳性对照组(顺铂浓度2.5μmol/L)、NCTD低、中、高剂量组(NCTD浓度分别为12.5、25、50μg/mL)。干预24 h后,划痕实验和侵袭实验检测PC-3细胞侵袭力的变化,透射电镜观察PC-3细胞的伪足形态变化,实时荧光定量PCR和Western Blot分别检测cortactin和MMP-9 mRNA与蛋白表达。结果各干预组PC-3细胞表面伪足样突起结构数量减少,随着NCTD浓度增加,伪足数量呈递减趋势。与模型组比较,各干预组细胞迁移距离、侵袭细胞数、cortactin及MMP-9 mRNA和蛋白表达降低(P<0.05);与阳性对照组比较,NCTD中、高剂量组细胞迁移距离、侵袭细胞数降低(P<0.05),NCTD高剂量组cortactin、MMP-9 mRNA及蛋白表达下降(P<0.05)。结论NCTD可降低PC-3细胞的侵袭能力,可能与下调cortactin、MMP-9蛋白表达,抑制PC-3细胞的伪足形成有关。

Objective To observe the effects of norcantharidin(NCTD)on migration,invasion and cortactin,MMP-9 mRNA and protein expression of PC-3 cells in prostate carcivoma.Methods The PC-3 cells were taken during the log-growth phase.They were divided into model group(no treatment),positive control group(cisplatin concentration 2.5μmol/L),low,medium and high dose NCTD group(NCTD concentrations were 12.5,25,50μg/mL,respectively).After 24h of intervention,invasion of PC-3 cells were observed by scratching and invasion experiments,pseudopodia morphological changes in PC-3 cells were observed by transmission electron microscopy,the mRNA and protein expression of cortactin and MMP-9 were determined by quantitative real-time PCR and Western Blot,respectively.Results The number of pseudopodialike protrusion structures of PC-3 cells in each intervention group decreased,as the NCTD concentration increased,the number of pseudopodia decreased.Compared with model group,cell migration distance,number of invasive cells,cortactin and MMP-9 mRNA and protein expression decreased in each intervention group(P<0.05).Compared with the positive control group,cell migration distance and the number of invasive cells were reduced in the middle and high-dose NCTD groups(P<0.05).Expression of cortactin,MMP-9mRNA and protein decreased in the high-dose NCTD group(P<0.05).Conclusion NCTD could decrease invasion ability of PC-3 cells,it may be related to down-regulation of cortactin and MMP-9 protein expression and inhibition of pseudopodia formation in PC-3 cells.