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-κB-hrGFP Plasmid from the PathDetect Signal Transduction Pathway cis-Reporting Systems Kit (Stratagene). These HT29-pNF-κB-hrGFP cells are routinely used at Cell Biology Unit (Institute Pasteur Montevideo - Uruguay) to screen natural or synthetic compounds that modulate NF-κB 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-κB activation assay, exponentially growing HT29 pNFκB-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-α, 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-α. 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-κB, a key component in the process of tumorigenesis.
Diogo Kuczera1 , Ines Tiscornia2 , Mariela Bollati-Fogollin2 , Dorly de F. Buchi1 1 – Department of Cell Biology, SCB, Universidade Federal do Paraná, Brazil. 2 – Cell Biology Unit (CBU), Institut Pasteur de Montevideo (IPMon), Uruguay