The Transfection Of Chitosan-shRNA Nanoparticles That Silence The Expression Of Stathmin Is Able To Enhance The Chemosensitivity Of Pulmonary Adenocarcinoma

Objaetives: To investigate whether transfection of the natural nano-material chitosan complexed with the shRNA expression plasmid targeting stathmin gene into A549 lung cancer cells could enhance the sensitivity of cells to chemotherapeutic drugs docetaxel, and provide a basis for chitosan nanoparticles as a vector in lung cancer gene therapy.Methods: ShRNA eukarya expression vector targeting stathmin gene was constructed and identified by restrictive analysis. Then the shRNA plasmid was transfected into the A549 cells,and the silencing efficiency of target gene was detected by RT-PCR and Western blot. Chitosan-shRNA nanoparticles targeting stathmin gene was formulated using a complex coacervation process. Flow cytometry was applied to analysis the cell cycle of A549 cells transfected with the nanoparticles. Chitosan-shRNA nanoparticles and cationic liposome-gene complex were transfected into A549 cells respectively, the sensitivity of cells to chemotherapeutic drugs docetaxel was detected by MTT assay.Results: ShRNA eukaryotic expression plasmid targeting stathmin gene called pGU6/GFP/Neo-shSTA was constructed successfully, the mRNA and protein expression levels of stathmin gene were significantly lower than the control group in transfected A549 cells (P<0.05);Chitosan-shRNA nanoparticles targeting stathmin gene was prepared successfully;GFP fluorescence expression of the reporter gene was detected in A549 cells transfected by nanoparticles;After transfected by nanoparticles, flow cytometry showed there was a G2 /M phase arrest in transfected cells (P<0.05);The value of IC50 in either chitosan-shRNA nanoparticles group or cationic liposome-gene complex group was significantly lower than in the control group(P<0.05), and the former is lower than the latter in the extent of reduction (P<0.05 ).Conclusion: The interference plasmid PGU6/GFP/Neo-shSTA can efficiently targetingly silence the expression of stathmin gene.Transfection of chitosan-shRNA nanoparticles targeting stathmin gene into pulmonary adenocarcinoma cells can significantly enhance the sensitivity of cells to chemotherapeutic drugs docetaxel.