Dissertation
Dissertation > Medicine, health > Oncology > Respiratory system tumors > Lung tumors

The Transfection of Chitosan-shRNA Nanoparticles That Silence the Expression of stathmin Is Able to Enhance the Chemosensitivity of Pulmonary Adenocarcinoma

Author SongYinFeng
Tutor HuangQi
School Huazhong University of Science and Technology
Course General thoracic surgery
Keywords Lung cancer Nano Chitosan shRNA stathmin Chemotherapy
CLC R734.2
Type Master's thesis
Year 2010
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Objective : To observe the the natural nanomaterials chitosan wrapped stathmin gene short hairpin RNA (shRNA) expression plasmid-mediated transfected lung adenocarcinoma cell A549, whether the enhanced sensitivity of the cells to the chemotherapy drug paclitaxel , chitosan nanoparticles contained genes provide the basis of the particles in the biological treatment of lung adenocarcinoma . Method : build short hairpin RNA (shRNA) targeting stathmin gene eukaryotic expression plasmid and digestion , and sequencing ; liposome -mediated transfection of A549 cells , RT-PCR , Western Blot method to detect the target genes of the plasmid silent performance ; using complex coacervation the stathmin the chitosan -shRNA nanoparticles synthesis targeting ; nanoparticles of chitosan -shRNA gene complexes of cationic liposome transfected into A549 cells , MTT assay cells to chemotherapeutic agents paclitaxel sensitivity changes . Results: successfully constructed the shRNA targeting stathmin gene eukaryotic expression plasmid pGU6/GFP/Neo-shSTA ; A549 cells transfected stathmin gene mRNA and protein expression levels compared with the control group were significantly lower ( P lt; 0.05 ) ; successful prepared nanoparticles of chitosan -shRNA targeting stathmin ; the nanoparticles in cells transfected A549 cells can be detected obvious expression of the reporter gene (GFP) ; chitosan -shRNA nanoparticles , cationic liposome gene complexes transfected A549 cells group compared with the control group , half inhibition (IC50) was significantly lower ( P lt; 0.05 ) , the chitosan group to reduce the degree of less than the liposome group ( P lt; 0.05 ) . Conclusion : Plasmid pGU6/GFP/Neo-shSTA efficiently targeted silencing stathmin gene expression ; experiments prepared chitosan nanoparticles contained genes particles , successfully transfected A549 cells ; targeting the stathmin of chitosan -shSTA of nanoparticles transfected lung adenocarcinoma cells , can significantly enhance the sensitivity of the cells to the chemotherapy drug paclitaxel ; chitosan mediated shRNA transfection enhanced sensitivity to paclitaxel in A549 cells was lower than liposomes .

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