Inhibition of matrix metalloproteinase-2 expression and bladder carcinoma metastasis by halofuginone.
Matrix metalloproteinase-2 (MMP-2) plays a critical role in tumor cell invasion and metastasis. Inhibitors of this enzyme effectively suppress tumor metastasis in experimental animals and are currently being tested in clinical trials. MMP-2 transcriptional regulation is a part of a delicate balance between the expression of various extracellular matrix (ECM) constituents and ECM degrading enzymes. Halofuginone, a low-molecular-weight quinazolinone alkaloid, is a potent inhibitor of collagen type alpha1 (I) gene expression and ECM deposition. We now report that expression of the MMP-2 gene by murine (MBT2-t50) and human (5637) bladder carcinoma cells is highly susceptible to inhibition by halofuginone. Fifty percent inhibition was obtained in the presence of as little as 50 ng/ml halofuginone. This inhibition is due to an effect of halofuginone on the activity of the MMP-2 promoter, as indicated by a pronounced suppression of chloramphenicol acetyltransferase activity driven by the MMP-2 promoter in transfected MBT2 cells. There was no effect on chloramphenicol acetyltransferase activity driven by SV40 promoter in these cells. Halofuginone-treated cells failed to invade through reconstituted basement-membrane (Matrigel) coated filters, in accordance with the inhibition of MMP-2 gene expression. A marked reduction (80-90%) in the lung colonization of MBT2 bladder carcinoma cells was obtained after the i.v. inoculation of halofuginone-treated cells as compared with the high metastatic activity exhibited by control untreated cells. Under the same conditions, there was almost no effect of halofuginone on the rate of MBT2 cell proliferation. These results indicate that the potent antimetastatic activity of halofuginone is due primarily to a transcriptional suppression of the MMP-2 gene, which results in a decreased enzymatic activity, matrix degradation, and tumor cell extravasation. This is the first description, to our knowledge, of a drug that inhibits experimental metastasis through the inhibition of MMP-2 at the transcriptional level. Combined with its known inhibitory effect on collagen synthesis and ECM deposition, halofuginone is expected to exert a profound anticancerous effect by inhibiting both the primary tumor stromal support and metastatic spread.
Elkin
M
M
Department of Oncology, Hadassah-Hebrew University Hospital, Jerusalem, Israel.
Reich
R
R
Nagler
A
A
Aingorn
E
E
Pines
M
M
de-Groot
N
N
Hochberg
A
A
Vlodavsky
I
I
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002277:Carcinoma / Q000201:enzymology* / Q000378:metabolism / Q000556:secondary; D003094:Collagen; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D005780:Gelatin / Q000378:metabolism; D006801:Humans; D007797:Laminin; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors; D051379:Mice; D008809:Mice, Inbred C3H; D009361:Neoplasm Invasiveness; D009368:Neoplasm Transplantation; D010880:Piperidines; D011401:Promoter Regions, Genetic / Q000187:drug effects; D011509:Proteoglycans; D011799:Quinazolines / Q000494:pharmacology*; D052999:Quinazolinones; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000201:enzymology* / Q000378:metabolism / Q000473:pathology
D000970:Antineoplastic Agents; D004338:Drug Combinations; D007797:Laminin; D061965:Matrix Metalloproteinase Inhibitors; D010880:Piperidines; D011509:Proteoglycans; D011799:Quinazolines; D052999:Quinazolinones; C059485:matrigel; D005780:Gelatin; D003094:Collagen; D020778:Matrix Metalloproteinase 2; C010176:halofuginone