Baculovirus Mediated Experimental Research on Targeted Egr1-Kringle 5 Gene Radiotherapy in Lung Adenocarcinoma英文文献资料.docVIP

Journal of Cancer Therapy, 2012, 3, 397-405 397 /10.4236/jct.2012.324052 Published Online September 2012 (http://www.SciRP.org/journal/jct) Baculovirus Mediated Experimental Research on Targeted Egr1-Kringle 5 Gene Radiotherapy in Lung Adenocarcinoma Haoping Xu, Rui Guo, Biao Li * Department of Nuclear Medicine, Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China. * Email: lb10363@ Received April 15 , 2012; revised May 27 , 2012; accepted June 15 , 2012 th th th ABSTRACT Objective: To investigate the feasibility of temporally and spatially restricted Kringle5 expression induced by radiation, as well as the dual effect of radiotherapy and antiangiogenic therapy in lung adenocarcinoma in vitro. Methods: We first constructed recombinant baculovirus vectors containing Egr1 promoter and human plasminogen Kringle5 gene (rhK5), then transfected them into lung adenocarcinoma cells (A549). Transfect efficiency of the baculovirus for gene transfer in A549 cells and the activity of Egr1 promoter induced by X-radiation were detected by fluorescence micros- copy. The rhK5 mRNA transcription and rhK5 protein expression were detected by Real-time PCR and Western blot assay, respectively. The apoptosis asssay of human umbilical veins endothelial cells (HUVEC) was analyzed by flow cytometry. Results: The recombinant baculovirus were successfully transfected into A549 and HUVEC cells. As for the temporal regulation, the rhK5 mRNA transcription and rhK5 protein expression were elevated with the irradiation time significantly. And the HUVEC apoptotic percentage increased in relation to the irradiation time as well. As for the spa- tial regulation, rhK5 mRNA transcription level of A549 cell lines transfected with recombinant baculovirus Egr1-K5 was significantly higher than that of control grou