RESUMO
BACKGROUND: Metastasis is the major cause of treatment failure in cancer patients and cancer- associated death, and an antimetastatic drug would be a beneficial therapy for cancer patients. HM-3-HSA is a fusion protein which improved the pharmacokinetics of HM-3 and exerted antitumor and anti-angiogenesis activity in multiple tumor models. However, the efficacy of HM-3-HSA in cancer cell migration and metastasis has not been elucidated. MATERIALS AND METHODS: Herein, high-cell density fermentation of Pichiapink strain expressing HM- 3-HSA was performed for the first time. Then, the desired protein was purified by Butyl Sepharose High performance, Capto Blue, Phenyl Sepharose 6FF HS and DEAE Sepharose FF. Furthermore, the effect of HM-3-HSA on the migration and invasion of cancer cells was also evaluated, and B16F10 metastasis model was established to detected the anti- metastasis effect of HM-3-HSA in vivo. RESULTS: The results indicated that the yield of HM-3-HSA was 320 mg/L in a 10 L fermenter, which was a 46% increase over that expressed in flask cultivation. The desired protein was purified by fourstep, which yielded a 40% recovery of a product that had over 99% purity. Purified HM-3-HSA significantly suppressed the migration and invasion of HCT-116, SMMC-7721 and B16F10 cell lines. CONCLUSION: On the other hand, in the B16F10 metastasis model, HM-3-HSA significantly inhibited pulmonary metastases of B16F10 cells, suggesting that HM-3-HSA exerted the anti-metastasis effect in vivo.
