Objective: To explore the role of neurogenin 3(NGN3) and paired box gene 4(PAX4) in the process of PDX1-driven mesenchymal stem cells (MSCs) to the pancreatic β-cell differentiation.
Methods: PDX1 gene and NGN3 were constructed with PAX4 genes expression adenovirus vectors, Adxsi-CMV-PDX1 adenovirus infection induced MSCs. One week later, re-Adxsi-CMV-NGN3/CMV-PAX4 adenovirus infection induced MSCs; and detected PDX1, PAX4, NGN3, NK transcription factor related, gene family 2, locus2(NKX2.2), v-maf musculoaponeurotic fibrosarcoma oncogene homolog B(MafB), insulin, glucagon and other pancreatic secretory cell-associated molecule expression.
Results: Adxsi-CMV-PDX1 and Adxsi-CMV-NGN3/CMV-PAX4 adenovirus vectors were constructed successfully. Through immuocytochemistry and indirect fluorescence detection, the expressions of PDX1 and NGN3 and PAX4 factors were detected stably in MSCs cellular nuclei which were induced by Adxsi-CMV-PDX1 and Adxsi-CMV-NGN3/CMV-PAX4. After transfection for 5 d, the cells formed round, gathered into a mass and showed bright red with dithizone staining. RT-PCR detection showed NruroD1 and NKX2.2 expression after being induced for 1 week and insulin/proinsulin molecules after being induced for 2 week. The induced cells could express some transcription factors such as NKX2.2 and MafB, and also pancreatic-secreting related molecules such as insulin and glucagon, but not the expressions of MafA and C-peptide.
Conclusion: NGN3 and PAX4 have a favourable role in PDX1 inducing mesenchymal stem cells into pancreatic secretory cells.