[Construction and application of a recombinant plasmid inducing cell death in hepatocytes after transfection].

Objective: The mechanisms of mesenchymal stromal cells (MSCs)-mediated treatment of liver damage have been unclear. Two major mechanisms, which involve paracrine effects and/or direct trans-differentiation, have been proposed. To clarify which mechanism is more important, we planned to construct a recombinant plasmid expressing green fluorescent protein (GFP) driven by the cytomegalovirus (CMV) promoter and Pseudomonas aeruginosa exotoxin 40 (PE40) driven by the albumin (alb) promoter, which can induce cell death as soon as MSCs differentiate into hepatocytes.
Methods: To construct the recombinant eukaryotic expression vector pFlag-CMV-GFP-TM-albp-PE40, GFP, transmembrane domain of DLL1, alb promoter and PE40 were obtained by PCR and were inserted into pFlag-CMV-1. The expression of GFP was observed under a fluorescence microscope and the killing effect on hepatocytes was analyzed by flow cytometry.
Results: The recombinant plasmid inducing cell death in hepatocytes was successfully constructed, suggesting that the plasmid could be employed to study the mechanism of MSCs-mediated treatment on liver damage.
Conclusion: This study might provide a promising tool for revealing the mechanism of MSCs-mediated treatment on liver diseases.