Human endostatin gene expression adenovirus vector and its expression in activity in breast cancer cells.docVIP

 PAGE \* MERGEFORMAT 15 Human endostatin gene expression adenovirus vector and its expression in activity in breast cancer cells Keywords: adenovirus gene therapy of endostatin anti-angiogenic treatment of breast cancer 0 Introduction In recent years, to prevent tumor angiogenesis, control of tumor growth and metastasis has become a new strategy for cancer therapy [1,2], which uses transfection angiogenesis inhibitor angiogenesis antagonistic to the ideal of endostatin [3 ]. In this paper, building human endostatin adenovirus gene expression vector to study endostatin expression in breast cancer cells for further study of breast cancer anti-angiogenic gene therapy ready. 1 Materials and methods 1.1 Materials Carrying human endostatin gene (human endostatin, hE) plasmid pBlast  hEndostatin purchased in the United States InvitroGen, Inc.; adenovirus vector pCA13, pBGHE3 and 293 cell lines were purchased in Microbix Biosystems, Inc.; human breast cancer cell line MCF and MBD  231  7, human umbilical vein endothelial cell line ECV  304 were purchased in the U.S. ATCC cell bank; LipofectAMINE2000 transfection kit purchased from GibcoBRL, Inc.; a variety of restriction enzymes and DNA ligase were purchased from NEB Inc.; QIAprep spin miniprepkit, QIAamp DNA Blood Mini Kit purchased from QIAGEN Inc.; M  PER Mammalian Protein Extraction Reagent were purchased from PIERCE companies; Western color liquid LumiGLO chemiluminescent reagent and peroxide for Cell Signaling Technology’s products; hE mouse anti-human monoclonal antibody, FITC labeled goat anti - mouse IgG antibody, HRP labeled goat anti-mouse IgG antibodies were purchased from Wuhan Boster Company; carrying LacZ reporter gene in the proliferation of defective adenovirus Ad  LacZ by the Eastern Hepatobiliary Surgery Hospital, viruses and gene therapy research to save. 1.2 adenovirus vector plasmid pAd  hE Construction According to GenBank AF18