STUDY ANTICANCER ACTIVITY FROM THE LIGHT SUBUNIT OF MUSHROOM TYROSINASE (LSMT) IN BREAST CANCER MODEL MDA-MB-231 CELL LINE AND DEVELOPMENT OF SPHEROID AS TUMOR MODEL

• Main Author: Azzahra, Fauzia
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/54335
• Institution: Institut Teknologi Bandung

Light Subunit of Mushroom Tyrosinase (LSMT) is a lectin-like protein from Agaricus bisporus, which was discovered coincidentally during elucidation of the crystal structure of the enzyme. Recombinant Light Subunit Mushroom Tyrosinase (rLSMT) successfully produced. rLSMT is a non-immunogenic and nonagglutinating protein. It has an anti-proliferative activity on MDA-MB-231 cell line. Aberrant signaling pathway occurs in cancer, such as Wnt signaling. Increase Wnt signaling pathway marked by overexpression of ?-catenin and its downstream target such as cyclin-D1. Cancer Stem Cells (CSC) are subpopulation of cancer cells which have self-renewal, proliferation, differentiation capabilities and initiate as well as sustaining tumor growth in vivo. The aim of this research is to explore the potential activity of rLSMT as anticancer protein through Wnt signaling pathway. In addition, we also developed 3-dimension culture, Spheroid, to analyze the tumorigenesis process. Spheroid will act as a model for drug activity screening, mimicking the clinical situation as compared to monolayer cell culture. Activity of rLSMT on regulation of Wnt signaling pathway in MDA-MB-231 cell line was observed due to its role in regulation of proliferation, differentiation, and selfrenewal. Spheroid was generated by culturing the MDA-MB-231 cell line in a nonadherent surface, supplemented with EGF, b-FGF and B27. No significant size reduction of MDA-MB-231 spheroid after treatment with 350 ?g/mL LSMT. Treatment with 579.3 ?g/mL of LSMT tent to reduce the expression of ?-catenin and cyclin D1 genes although Wnt signaling was not the major mechanism. Overall, we conclude that 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. In other site, the successful development of spheroid provides opportunity for further study of anticancer drugs in vitro system which is mimicking the clinical condition of cancer.