Antitumoral and antimetastatic activity of maitake D-fraction in triple-negative breast cancer cells

ALONSO E.N.; FERRONATO M.J.; GANDINI N.A.; FERMENTO M.E.; GUEVARA J.A.; MARIANI F.; FACCHINETTI M.M.; CURINO A. C.

Buenos Aires

Congreso; Reunión Conjunta de Sociedades de Biociencias. LXII Reunión Anual de la Sociedad Argentina de Investigaciones Clínicas (SAIC); 2017

Sociedad Argentina de Investigaciones Clínicas (SAIC)

D-Fraction is a  proteoglucan  extracted from  Grifola frondosa  (Maitake)  mushroom.Previously, we reported  that  D-Fraction decreases breast cancer  (BC)  cell viability regardless  of  hormone  receptors  and  HER2  status  of  cells.  Furthermore,  DFraction  reduces  tumor  burden  and  lung  metastases  in  a  murine  model  with hormone-independent LM3  cells.  In triple-negative  (TNBC) MDA-MB-231 cells,  we also demonstrated that D-Fraction decreases their migration and invasion capacity. The  purpose  of  the  current  study  is  to  identify  the  cellular  and  molecular mechanisms  by  which  D-Fraction  decreases  the  migratory/invasive  potential  of  MDA-MB-231  cells.  In addition, we  propose to evaluate the antitumoral effect of DFraction in 4T1 cells, another cell line representative  of  TNBC subtype.  By western blot,  we  found  that  D-Fraction  increases  E-cadherin  expression  in  MDA-MB-231 cells  compared  to  vehicle  treatment  (p<0.05).  By  immunofluorescence,  we detected  that  D-Fraction  decreases  the  presence  of  â-catenin  in  the cytoplasm/nucleus  (p<0.001)  and  promotes  its  membrane  localization  (p<0.01). Also,  we  found  that  D-Fraction  increases  the  adhesion  of  MDA-MB-231  cells  to substrate (p<0.05). By zymography, we detected that D-Fraction decreases MMP-2 and  MMP-9  activity  by  53.59  %  (p<0.001)  and  27.31  %  (p<0.05)  respectively, compared  to vehicle treatment. On the other hand, manual cell counting and WST-1 assay were performed in TNBC 4T1 cells.  D-Fraction decreases the viability of 4T1 cells in a dose-  and time-dependent manner  (p<0.05).  Wound healing assay demonstrated  that  D-Fraction  decreases  the  migratory  capability  of  4T1  cells  (p<0.001). By  transwell Matrigel  assay,  D-Fraction reduces  the  invasive  capability of  these  cells  (p<0.001).  In  conclusion,  our  results  suggest  that  D-Fraction decreases  the  viability  and  metastatic  potential  of  TNBC  cells:  promoting  an epithelial phenotype; reducing  the  capability  of tumor cells  to degrade extracellular matrix and increasing cell-substrate adhesion.