Avaliação biológica in vitro da lectina BvL e a isoforma recombinante I (rBvL-I) sobre a viabilidade de linhagens celulares epiteliais.

Abstract

Lectins are a group of proteins with several particular characteristics, one of which is the ability to stimulate cell proliferation. They are proteins that recognize carbohydrates in a specific and reversible way. In this work we tested the BvL native lectin from Bauhinia variegata and a recombinate isoform rBvL-I. The Bauhinia variegata plant belongs to the Leguminosae family, where there are more than 70 lectins already reported. Native nBvL lectin is isolated from the seed of this plant and has approximately 33 kDa due to glycosylation, whereas recombinant isoform I, rBvL-I, has approximately 28 kDa. The ability of these proteins to induce cell proliferation was evaluated in cell cultures involved in the tissue regeneration process, such as human fibroblasts (HFF-1), human keratinocytes (HaCaT) and mouse murine fibroblasts (NIH / 3T3). Lectin has also been tested on human skin cancer melanoma cells (A-375), in order to verify a possible action on these cells. Several tests were carried out to assess cell viability and proliferation of lectin in these cells, such as the MTT assay, flow cytometry with the EdU assay and cell count assay, confocal microscopy and Western blot. It was possible to verify the stimulus of the proliferation of human fibroblasts and murine fibroblasts.