Expressão de uma serinoprotease de Crotalus durissus collilineatus em células de mamíferos e avaliação da sua atividade citotóxica sobre diferentes linhagens tumorais.

Abstract

BACKGROUND: In the last decades, studies reporting the action of ophidian toxins with therapeutic potential have increased significantly and several new compounds have been tested, showing improvement in some pathologies, mainly on some types of cancers. Cancer of the head and neck is known as having anatomical features that define malignant tumors of the upper digestive tract. Taking into account the global increase in cancer cases, it is necessary to search for alternatives to inhibit tumor progression, as well as the development of efficient drugs. Some isolated or even recombinant toxins have been found to be a good alternative for the treatment of tumor cells. Among a range of toxins present in the venom of Crotalus durissus collilineatus are serine proteases, which act on plasma proteins associated with the blood coagulation pathway. Snake venoms are a vast source of pharmacologically active biomolecules. Despite the potential for exploration, there are few studies on the biological characterization of serine proteases with antitumor action. AIM:The objective of this work was cloning and expression of a serine protease present in the venom of Crotalus durissus collilineatus and evaluate its therapeutic potential in some cell lines derived from head and neck tumors, breast and lung cancer. MATERIALS AND METHODS: The sequence of serine protease was inserted into the pCI-Neo expression plasmid. The potential of the cell line constructed expressing the protein was tested in cell lines derived from head and neck tumors, as well as lineages derived from breast and lung by techniques such as Western Blot, MTS and apoptosis RESULTS: The expression of serine protease was confirmed by the Western blot assay, which was constructed from a transfection with the pCI-Neo expression plasmid containing the sequence of serine protease, and the effect of the same on the viability of the lineages tumors of the head and neck, breast and lung. Although preliminary assays made from a transient transfection of the pCI- Neo expression plasmid containing the serinoprotease showed a decrease in cell viability in the VU147T cell line. when the cell line was established, it did not present statistical significance in any of the tumor lines that were treated with the supernatant of the cell line with constitutive expression of the serinoprotease. CONCLUSION: few studies are found in the literature that report the action of these proteases on tumor cells. Thus, the studies presented in this study revealed the efficiency in the production of the recombinant serine protease of C. d. collilineatus, but further testing is needed to understand the mechanism of action of these proteases against tumor cells.