J Pharm Pharmacogn Res 3(suppl. 1): S95, 2015
Proceedings of the 4th International Symposium on Pharmacology of Natural Products FAPRONATURA 2015 September 21st-25th, 2015; Cuban Society of Pharmacology. Topes de Collantes, Sancti Spiritus, Cuba.
PPP-03: PROTEOLYTIC FRACTION FROM Vasconcellea cundinamarcensis LATEX REDUCES ACTIVITY OF THE MATRIX METALLOPROTEINASE DURING MELANOCYTE TRANSFORMATION in vitro
Santos VG1, Lemos FO1, Lage FDO1, Salas CE2, Lopes MTP1.
Introduction: CMS2 is a proteolytic fraction from Vasconcellea cundinamarcensis latex obtained after chromatographic fractionation. Previous studies showed that this fraction presents antimetastatic activity on murine melanoma and colon carcinoma models. This effect may be related to an increase in DNA fragmentation, along with a reduction of cell adhesion and invasion. Aim: The present study evaluates the action of CMS2 on metalloprotease activity, used as marker during melanocyte (Melan-a, nontumorigenic murine cell line) differentiation undergoing cell transformation by sequential cycles of anchorage blockage. Methods and Results: The Melan-a cell viability, exposed to CMS2 (1-50 µg/mL) for 24 h, was assessed by metabolization of rezasurin (reading 570/600 nm). The assay showed a 50% cell inhibitory concentration (IC50) at 5 µg/mL. Next, Melan-a cells were pretreated with CMS2 (1-10 µg/mL) for 12 h and seeded (105 cells/mL) on 1% agarose. The resulting spheroids, after 96 h, were harvested and seeded on culture flasks, to favor cell adhesion (procedure repeated five times). In cells obtained in each cycle, the secreted metalloprotease 9 (MMP-9) activity was determined by zymography. The densitometric analysis of gels (software ImageJ® 1.46) showed that treatment with 5 – 10 µg/mL CMS2 reduced the activity of MMP-9 until 94% (5.54 ± 0.95) relative to the untreated group (100.00 ± 11.38, p<0.0001, one-way ANOVA test, Student-Newman-Keuls post-test). Conclusions: These results suggest that CMS2 can prevent the melanocytes transformation, by the reduction of MMP-9 activity, which consequently can affect the ability of cell invasion, since MMP-9 is closely linked to this process. Financial Support: CNPq, CAPES and FAPEMIG.
Citation Format: Santos VG, Lemos FO, Lage FDO, Salas CE, Lopes MTP (2015) Proteolytic fraction from Vasconcellea cundinamarcensis latex reduces activity of the matrix metalloproteinase during melanocyte transformation in vitro. [Abstract]. In: Proceedings of the FAPRONATURA 2015; 2015 Sep 21-25; Topes de Collantes, Sancti Spiritus: CSF. J Pharm Pharmacogn Res 3(Suppl. 1): S95. Abstract nr PPP-03.