M1 As A Promising Product That Modulates NF-kB Activation
Dorly de F. Buchi
Department of Cell Biology, SCB, Universidade Federal do Paraná, Brazil
Abstract:
The family of NFkB transcription factors has a critical role in cancer development and progression, regulating tumor angiogenesis and invasiveness. It is an attractive target for new chemopreventive and chemotherapeutic approaches. The aim of this work is to develop highly diluted natural complexes with anti-cancer properties without side effects. One of them is a complex matrix from Chelidonium and associations, coded as M1. Previous results using those complexes in our laboratory have shown immunomodulatory and anti-melanoma activity in vitro and in vivo. Human colon-rectal cancer cells (ATCC: HTB-38) were stably transfected with the pNF-?B-hrGFP Plasmid from the PathDetect Signal Transduction Pathway cis-Reporting Systems Kit (Stratagene). These HT29-pNF-?B-hrGFP cells are routinely used at Cell Biology Unit (Institute Pasteur Montevideo - Uruguay) to screen natural or synthetic compounds that modulate NF-?B activity. Treatment was administered to log-phase growing cells. An initial dose of 20% of M1 was administered to the cells and, after 24h a reinforcement dose of 1% was added. In the NF-?B activation assay, exponentially growing HT29 pNF-?B-hrGFP cells (5x105/ml) were seeded in 96 well-plate and grown for 24 h. The cells were then cultured for 48 h in presence or absence of 3 ng/mL TNF-a, with or without M1 complex. Twenty thousand events were acquired, analyzed and the percentage of positive GFP cells was determined using CyAn™ ADP Flow Cytometer and Summit v4.3 software. After 48h we found a significant (p<0.05) reduction in the number of cells expressing GFP when treated with M1 in presence of TNF-a. This result opens directions for new researches using M1 complex as possible adjuvant in the therapy of cancer once it has shown to be interacting and decreasing NF-?B, a key component in the process of tumorigenesis.
