Tbx20 gene overexpression increases cell proliferation and the levels of pro mitotic genes in post-natal rat cardiomyocytes.

BAUZÁ MARÍA DEL ROSARIO; HALEK JULIA MARÍA; CROTTOGINI ALBERTO; SIMONIN JORGE ALEJANDRO; CUNIBERTI LUIS; OLEA FERNANDA DANIELA; LÓPEZ AYELÉN EMILCE; BELAICH MARIANO NICOLÁS; LOCATELLI PAOLA

Rosario

Congreso; Reunión Anual SAFIS Rosario 2019; 2019

Sociedad Argentina de Fisiología

Introduction: myocardiogenic therapies have been proposed to induce cardiac regeneration after myocardial infarction. One of possible approaches would be to stimulate the adult cardiomyocyte to re-enter the cell cycle and divide into daughter cells. To achieve this, positive and negative cell cycle regulator molecules could be down regulated. The induced overexpression of TBX20 transcription factor in transgenic mice increased cell proliferation. Objective: To evaluate the cell proliferation and gene expression of the cell cycle stimulator genes (Cyclin D1 and MEF2C) and repressor gene (p21) in post-natal rat cardiomyocyte culture. Methods: Baculoviral vectors encoding Tbx20 (BvTbx20) and GFP (BvGFP) genes were generated. Post-natal cardiomyocytes (CM) were cultured and transduced at 3 days of culture with BvGFP at different multiplicities of infection (MOI: 25, 50, 100, 200 and 500). Transduction efficiency was measured by flow cytometry. Cell proliferation (MTS assay) and gene expression of Cyclin D1, MEF2C and p21 (RT-qPCR) were measured in CM transduced with BvTbx20 at different days.Results: Transduction efficiency was optimal at MOI 100 (41.72%). Percentage of cell proliferation was higher in transduced CM (Tbx20CM: 109.6±4.5) respect to non transduced CM (ntCM: 100±3.2, p