J Pharm Pharmacogn Res 3(suppl. 1): S96, 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-04: PROTEOLYTIC FRACTION FROM Vasconcellea cundinamarcensis LATEX PROMOTES MACROPHAGE TUMORICIDAL ACTIVITY AGAINST BREAST CANCER CELLS
Braga AD1, Freitas KM1, Teixeira LCR1, Salas CE2, Lopes MTP1.
Introduction: Previous studies demonstrated that a proteolytic fraction (P1G10) from Vasconcellea cundinamarcensis latex has antitumor/antimetastatic activity on 4T1 breast carcinoma model by reducing inflammation, angiogenesis and increasing tumor associated-macrophages (TAMs) activity. Moreover, macrophages exposed to P1G10 are capable to reduce 4T1 cells number in co-culture model. Aim: Investigate the ability of P1G10 proteolytic sub-fractions (CMS1 and CMS2) to promote macrophage tumoricidal phenotype and its mechanisms of activation. Methods and Results: Balb/c female mice macrophages were harvested by peritoneal laved after received thioglycollate (3%, i.p., 2 mL, 3 days) (CETEA 219/2012). These cells were seeded (3 x 105 cells/well) and exposed to sub-fractions (10-40 µg/mL, 24 h). Treatments were removed and co-cultures made by 4T1 tumor cells addition (105 cells). After 40hrs, by MTT metabolization, 4T1 cells viability was determined (O.D. co-culture – O.D. macrophage culture). Just macrophages pre-exposed to CMS2 (20-40 µg/mL) reduced 4T1 cells viability until 57% (0.36±0.02 ΔO.D., control, p<0.0001).The fluorescence(480Ex–530Em nm) was measured as ROS production in macrophages exposed to sub-fractions (5-40 µg/mL, 24 h) and labeled with DCFH-DA (15 µM, 30 min). CMS1 and CMS2 increased ROS production until 104% and 171%, respectively (8,613.00 ± 103.00 ΔFU/cell number, control, p< 0.0001). Both sub-fractions (20-40 µg/mL), increased nitric oxide (NO) production, determined indirectly using Griess method, reaching until 969.00% (CMS1) and 2,249.00% (CMS2) (0.09 ± 0.02, control, p< 0.0001). Statistical analysis: One-way ANOVA test, Student-Newman-Keuls post-test. Conclusions: The results suggest that ROS and NO are involved in macrophage cytotoxic activity induced by P1G10 and that the proteins contained in CMS2 sub-fraction are mainly responsible for this effect. Financial Support: CNPq, Capes and Fapemig.
Citation Format: Braga AD, Freitas KM, Teixeira LCR, Salas CE, Lopes MTP (2015) Proteolytic fraction from Vasconcellea cundinamarcensis latex promotes macrophage tumoricidal activity against breast cancer cells. [Abstract]. In: Proceedings of the FAPRONATURA 2015; 2015 Sep 21-25; Topes de Collantes, Sancti Spiritus: CSF. J Pharm Pharmacogn Res 3(Suppl. 1): S96. Abstract nr PPP-04.