Topotecan inhibits cancer cell migration by down-regulation of chemokine CC motif receptor 7 and matrix metalloproteinases.
OBJECTIVE
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 CCR7 in both MDA-MB-435 and MDA-MB-231 cells and moderately reduced the expression of CXCR4 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.


CONCLUSIONS
TPT inhibited cancer cell migration by down-regulation of CCR7 and MMPs (MMP-2 and MMP-9).
National New Drug Screening Lab, China Pharmaceutical University, Nanjing, China.
Sen-sen
SS
Lin
Li
L
Sun
Yan-kai
YK
Zhang
Ren-ping
RP
Zhao
Wen-lu
WL
Liang
Sheng-tao
ST
Yuan
Lu-yong
LY
Zhang
D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D015536:Down-Regulation / Q000187:drug effects*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D009362:Neoplasm Metastasis / Q000188:drug therapy / Q000517:prevention & control; D054400:Receptors, CCR7 / Q000235:genetics / Q000378:metabolism*; D019718:Receptors, CXCR4 / Q000235:genetics / Q000378:metabolism; D019772:Topotecan / Q000494:pharmacology* / Q000627:therapeutic use; D015854:Up-Regulation
D054400:Receptors, CCR7; D019718:Receptors, CXCR4; D019772:Topotecan; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9