Acta Pharmacologica Sinica (2009) 30: 628-636; doi: 10.1038/aps.2009.32; published online 13th April 2009

Aim: The aim of this study was to investigate the effect of topotecan (TPT) on cancer cell migration. 

Methods: Growth inhibition of TPT was analyzed by MTT assay, and cancer cell migration was measured by transwell double chamber assay.  To verify the effect of TPT on the chemokine receptors CXCR4 and CCR7, quantitative PCR, semi-quantitative PCR and Western blot analysis were performed.  The secretion of MMP-2 and MMP-9 was detected by enzyme-linked immunosorbent assay (ELISA) and gelatin zymography.  To evaluate possible contributions of CCR7 to MMP secretion, the overexpression vectors pcDNA3.1⁺-CCR7 and CCR7 siRNA were transiently transfected into MDA-MB-435 cells.

Results: TPT inhibited cancer cell migration in a dose-dependent manner.  Additionally, TPT significantly decreased the expression of CCR 7 in both MDA-MB-435 and MDA-MB-231 cells and moderately reduced the expression of CXCR 4 in MDA-MB-435 cells.  The secretion of MMPs (MMP-2, MMP-9) was also inhibited by TPT.  Overexpression of CCR7 increased the secretion of MMP-2/9 and cancer cell migration, whereas knockdown of CCR7 reduced active MMP-2/9 production and migration of MDA-MB-435 cells.

Conclusion: TPT inhibited cancer cell migration by down-regulation of CCR7 and MMPs (MMP-2 and MMP-9).

Keywords: topotecan; MDA-MB-435; MDA-MB-231; cancer metastasis; CXCR4; CCR7; MMP-2; MMP-9