去甲斑蝥素抑制蛋白激酶C影响乳腺癌细胞侵袭转移的研究

目的探讨去甲斑蝥素（norcantharidin,NCTD）抑制乳腺癌细胞侵袭与转移的机制。方法应用迁移实验、趋化实验、transwell转移模型分别检测NCTD对不同侵袭力乳腺癌细胞株MCF-7、SKBR3和MDA-MB231的粘附、迁移和侵袭的影响;用Western blot检测NCTD作用前后乳腺癌细胞PKCζ表达的变化。结果 SKBR3和MDA-MB231的趋化、侵袭能力明显高于MCF-7;用NCTD处理后,三种乳腺癌细胞体外迁移、粘附和侵袭均受到不同程度抑制（P〈0.05）;NCTD对SKBR3和MDA-MB 231细胞迁移、侵袭抑制作用明显高于MCF-7细胞株（P〈0.05）,SKBR3和MDA-MB231细胞之间差异则无统计学意义（P〉0.05）。使用PKCζ抑制剂myristolated pseudosubstrate（PSζ）可促进NCTD抑制SKBR3和MDA-MB231的侵袭和迁移能力;经NCTD处理后,三种乳腺癌细胞PKCζ表达均降低。结论去甲斑蝥素可明显抑制人乳腺癌细胞株的细胞侵袭和迁移能力,其机制可能通过抑制PKCζ信号传导途径实现,与PKC抑制剂联用,可以增强治疗肿瘤的疗效。

Anti-Invasive and Anti-Metastasis Effect of Norcantharidin on High-Metastatic Human Breast Cancer Cell Lines

Objective To explore the inhibitory effect of norcantharidin （NCTD） on invasive and metastatic abilities of human breast cancer cell lines and its possible mechanisms.Methods Effects of norcantharidin among different human breast carcinoma cells migration was detected with wound-healing assay; its adhesion potential was tested with cell adhesion assay, and on invasive and migratory capacities were measured with Transwell matrigel invasion assay and Transwell motility assay, respectively.Expression of protein kinase C zeta（PKCζ） in breast tumour cells was detected by Western blotting.Results The in vitro invasion and migration ability of SKBR3 and MDA-MB 231 was higher than that of MCF-7 breast cancer cell line （P0.05）.There was no difference between SKBR3 and MDA-MB 231 cell lines （P0.05）.After treated with norcantharidin the invasion and motility ability of three breast cancer cell lines were obviously decreased（P0.05）.However, the invasion and proliferation ability of MCF-7 cell line was still weaker than those of SKBR3 and MDA-MB 231 breast cancer cell lines （P0.05）; and there was also no difference between the latter two cell lines （P0.05）.Conclusion Norcantharidin can down regulate the cell migration and invasion ability in breast cancer lines.The possible anticancer mechanisms might be relate to the signal transduction pathways of PKCζ.The effect of norcantharidin and PKCζ inhibitor myristolated pseudosubstrate （PSζ） might be synergetic to inhibit the invasion and proliferation ability of human high metastatic breast cancer cells.