Abstract

Abstract : Light Subunit of Mushroom Tyrosinase (LSMT) is a lectin-like protein from Agaricus bisporus and its recombinant successfully produced. rLSMT has an anti-proliferative activity on MDA-MB-231 cell line. Aberrant signaling pathway occurs in cancer, such as Wnt signaling that is marked by overexpression of β-catenin and downstream target such as cyclin-D1. The aim of this research is to explore the potential activity of rLSMT as an anticancer through modulating the Wnt signaling pathway and to develop 3-dimension culture, Spheroid, as an advanced in vitro model mimicking the solid tumor. Treatment MDA-MB-231 cell line on monolayer culture with 579.3 μg/mL LSMT tent to reduce the expression of β-catenin and cyclin D1 genes although Wnt signaling was not the major mechanism. Spheroid was generated by culturing the MDA-MB-231 cell line in a non-adherent surface, supplemented with EGF, b-FGF and B27. Reduction size of MDA-MB-231 spheroid after treatment with 350 μg/mL LSMT is negligible and needs further investigation. Overall, anticancer activity of rLSMT in MDA-MB-231 cell line through Wnt signaling is weak, therefore suggested not as the major mechanism of the LSMT anticancer activity. Besides, the development of spheroid provides opportunity for further study of anticancer drugs in vitro which is mimicking the clinical condition of cancer.