title: Activity and tumor specificity of human heparanase gene core promoter.
abstract: Heparanase (HPSE) plays a critical role in tumor metastasis and vascularization.             In addition, the human HPSE promoter has been cloned and characterized. However,             the activity and specificity of the HPSE promoter in tumor cells remains unclear.             The core fragment of the HPSE promoter was amplified and cloned into the multiple             cloning site of the pEGFP-1 vector. The recombinant plasmid pEGFP-Hp was transfected             into human umbilical vein endothelial cells (ECV304) and human hepatoma carcinoma             (HepG2), laryngocarcinoma (Hep2) and chronic myelogenous leukemia (K562) cell             lines. The vectors pEGFP-1 and pEGFP‑N1 were used as negative and positive controls,             respectively. The activity and expression of green fluorescent protein (GFP) were             analyzed. Results showed that the sequence of the amplified HPSE promoter was             in agreement with the GenBank data. The recombinant plasmid pEGFP-Hp was consistent             with the expected result. No GFP expression was observed in the transfected cells             in the pEGFP-1 group, but a high expression was observed in the pEGFP-N1 group.             As regards the pEGFP-Hp group, less fluorescence was revealed in ECV cells with             a relatively high fluorescence in tumor cells. The average transfection efficiencies             of pEGFP-Hp in the ECV304, HepG2, Hep2 and K562 cell lines were 3.9, 21.3, 10.8             and 6.5%, respectively, while those of pEGFP-Nl were 17.1, 24.0, 14.0 and 11.0%,             respectively. The HPSE gene promoter drives the expression of downstream genes             in a eukaryotic vector, specifically in tumor cell lines, but its activity is             relatively weak.
authors: Department of Hepatobiliary Surgery, Affiliated Yijishan Hospital of Wannan Medical College, Wuhu, Anhui 241001, PR China. drcxp@sohu.com
authors: Xiao-Peng
authors: XP
authors: Chen
authors: Liang-He
authors: LH
authors: Hu
authors: Wei
authors: W
authors: Cui
authors: Dong
authors: D
authors: Wang
authors: Ming-Shi
authors: MS
authors: Wu
authors: Jun-Sheng
authors: JS
authors: Luo
authors: Min
authors: M
authors: Zhong
mesh_terms: D001483:Base Sequence; D005822:Genetic Vectors / Q000378:metabolism; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D056945:Hep G2 Cells; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D020014:K562 Cells; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D000099175:Heparanase
chemical_list: D005966:Glucuronidase; D049452:Green Fluorescent Proteins; D011993:Recombinant Fusion Proteins; D000099175:Heparanase