¿COULD MAITAKE D-FRACTION INHIBITS THE MALIGNANT

ELIANA ALONSO; DIEGO OBIOL; GABRIELA BALOGH

Rosario

Congreso; Reunión Internacional de Ciencias Farmacéuticas. RICIFA; 2012

¿cOULD Maitake D-Fraction INHIBITS THE MALIGNANT PHENOTYPE IN breast cancer?   Alonso E1 (ealonso@criba.edu.ar), Obiol D1(dobiol@criba.edu.ar) and Balogh G1*(gbalogh@criba.edu.ar).   Laboratorio de Hongos Comestibles y Medicinales, Centro Científico Tecnológico, CERZOS-CONICET1.Camino La Carrindanga Km7, Bahía Blanca- CP 8000, Buenos Aires, Argentina.     Introduction   D-Fraction is a standardized form of isolated b-glucan polysaccharide and protein. This fraction is obtained both the mycelia and fruit body of Basidiomycete Grifola frondosa (Maitake) (1). Maitake D- Fraction has demonstrated the most potent immune enhancement and antitumor, resulting in the highest reduction rate in cancer proliferation (2,3). Also, it has been shown to have an antitumor effect on tumor-bearing mice (2). Is already known that D-Fraction acts through activation of various immune effectors that targeting tumor cells, including macrophages (4,5,6), NK (4,6,7), LTh1 (8,9,10), LTc (5) and denditrics cells (10). Moreover, Fraction D acts directly on the tumor cell, inducing apoptosis (11,12, 13), and modulating specifics molecular targets of the neoplasic cell (13). However, the exact molecular mechanism of Maitake anti-tumoral effect is still unclear. At the present work we are identifying which genes are responsible for the suppression of the tumoral phenotype induced by Maitake D-Fraction in breast cancer cells.   Materials and Methods   The bioactive D-Fraction, was provided by Mushroom Wisdom Inc, N.J, USA. Human breast cancer MCF-7 cells were treated with increased concentrations of Maitake D-Fraction (0, 36, 91, 183, 367 µg/ml) during 24 hours. Total RNA were isolated and cDNA Microarrays were hybridized containing 25,000 human genes. The resulting images were analyzed and normalized employing the Nexus Expression Software (Biodiscovery, CA). Samples were evaluated by the analysis of variance test. A difference between experimental groups was analyzed by Student’s t-test, p < 0.05.   Results   We found that D-Fraction treatment at 183 µg/ml significantly increase (log2 ratio ≥ 2.0) the expression of 430 genes in MCF-7 cells after 24 hours. Also, we detected that 75 genes were down-modulated in a dose dependent manner after Maitake D-Fraction treatment. Interestly, we found that 91µg/ml of D-Fraction during 24 hours induce the maximum down-modulation (log2 ratio ≤ -0.52). After investigating in previously published data the relationship between those genes and neoplasic disease, specifically breast cancer, we could elucidate the molecular mechanism induced by Maitake in the anti-tumoral action. We postulated that Maitake D-Fraction treatment block tumor cell growth and proliferation by overexpresion of p27/Kip1, RBBP4, RASSF2, CAV1, activation of TGF-b signaling and by downregulation of AKT, IGF y NfkappaB pathways in MCF-7 cells. Moreover, we found that D-Fraction treatment increases pro-apoptotic gene activation such as: BAK1, BCLAF1, BCL2L13, RASSF2, FADD y SPARC; while suppress activation of antiapoptotic PI3K-AKT signaling. Also, we can sugest that D-Fraction treatment plays a role blocking the metastatic nature of MCF-7 cells by inducing the over-expression of ITGA2, ITGA9, MTSS1 and down-modulation of CD44 and ICAM-3. Moreover, Maitake D-Fraction could be a potential agent to induce chemotherapy-multidrug sensitivity in breast cancer by overexpression of SPARC, CUL3 or by downmodulation of ABCG2.   Conclusions   In conclusion, our data support the concept that Maitake D-Fraction can modulate the expression level of molecular tumoral target, and thus could be able to control the breast cancer phenotype and even could be able to revert the aggressive/malignant phenotype by the molecular mechanisms postulated here.         References   Mayell M. Maitake extracts and their therapeutic potential–A Review. Altern Med Rev. 2001. 6(1):48-60. Hishida I, et al. Antitumor activity exhibited by orally administered extract from fruit body of Grifola frondosa (Maitake). Chem Pharm Bull. 1988. 36 (5):1819-1827. Nanba H. Antitumor activity of orally administered D-fraction from Maitake mushroom. J Naturopathic Med. 1993.1:10-15. Kodama N, et al. 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