PPP-04: PROTEOLYTIC FRACTION FROM Vasconcellea cundinamarcensis LATEX PROMOTES MACROPHAGE TUMORICIDAL ACTIVITY AGAINST BREAST CANCER CELLS

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.

Poster

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.

Departments of 1Pharmacology and 2Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais, Belo Horizonte, MG, Brazil. E-mail: cesbufmg@yahoo.com

 

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.