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Study of the Interaction between Akt and Islet-1/GCN5 Protein Complex during Mesenchymal Stem Cells Specific Differentiation into Cardiomyocytes
Chen Xueni, Zhu Jing*, Tian Jie, Yi Qin, Huang yue
The Children’s Hospital Chongqing Medical University, Ministry of Education Key Laboratory of Developmental Disease in Childhood and Chongqing Key Laboratory of Children Urogenital Development and Tissue Engineering, Chongqing 400014, China
Abstract: This study has investigated the interaction between Akt and Islet-1 in GCN5 (general control of amino acid biosynthesis protein 5) protein complex during C3H10T1/2 cells specific differentiation into cardiomyocytes， and further clarified the regulatory mechanism of differentiation of mesenchymal stem cells (C3H10T1/2). The mouse marrow mesenchymal stem cells C3H10T1/2 was infected with the lentiviral vector containing Islet-1 gene. The morphology of cells was observed by microscope， mmunofluorescence was applied to detect the levels of Islet-1， cTnT (cardiac troponin T)， Connexin43， coimmunoprecipitation (CoIP) was applied to detect the interaction between Akt and Islet-1-GCN5 protein complex. The morphology of C3H10T1/2 cell group infected with Islet-1 lentivirus showed cardiomyocyte-like cell， and the protein levels of Islet-1， cTnT and Connexin43 were higher than that in the uninfected group. At the peak time (the third week) of the protein level of Islet-1， the binding capacity of Islet-1 and GCN5 was lower and higher than that of the untreated group under the situation of activation and inhibition of Akt， respectively (P<0.05). The results suggested that when Islet-1 promotes cells into cardiac-specific differentiation process， the activity of Akt is antagonistic to the protein binding capacity of Islet-1 and GCN5 during induction of mesenchymal stem cell C3H10T1/2 into cardiomyocyte cells.
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