title: Downregulation of heparanase by RNA interference inhibits invasion and             tumorigenesis of hepatocellular cancer cells in vitro and in vivo.
abstract: Heparanase is an endoglycosidase that degrades heparan sulfate, the main             polysaccharide constituent of the extracellular matrix and basement membrane.             The expression of heparanase is associated with invasion, as well as the angiogenic             and metastatic potential of diverse malignant tumors. We used RNA interference             strategies to evaluate the role of human heparanase in a liver cancer cell line             and to explore the therapeutic potential of its specific targeting. Using an online             siRNA tool, we designed three small interfering RNA sequences to target the heparanase             coding region and cloned them into the pGenesil-1 vector. The siRNA vectors were             transfected into HepG2 liver cancer cells. Heparanase expression was measured             by real-time RT-PCR and Western blotting. Cell proliferation was detected by MTT             staining and plate colony formation. Cell cycle analysis was performed by flow             cytometry. In vitro invasion was measured by Matrigel invasion assay. We also             analyzed tumorigenicity in heparanase-suppressed HepG2 cells in nude mice. We             found that siRNA-1 (1214-1232) and siRNA-3 (611-629) targeting heparanase significantly             downregulated the expression of heparanase in HepG2 liver cancer cells. Compared             with its controls, siRNA-1 or siRNA-3 vectors efficiently inhibi-ted the proliferation             and invasion of HepG2 liver cancer cells in vitro and tumorigenesis in vivo. These             results suggest that heparanase-specific RNA interference has potential value             as a novel therapeutic agent for human liver cancer.
authors.affiliation: Institute of Gastroenterology, Southwest Hospital, Third Military Medical University, Chongqing 400038, PR China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Xiong
authors.forename: Mu-Han
authors.initials: MH
authors.lastname: Lü
authors.forename: Ya-Han
authors.initials: YH
authors.lastname: Fan
authors.forename: Ya-Ling
authors.initials: YL
authors.lastname: Cao
authors.forename: Chang-Jiang
authors.initials: CJ
authors.lastname: Hu
authors.forename: Yu-Yun
authors.initials: YY
authors.lastname: Wu
authors.forename: Shu-Min
authors.initials: SM
authors.lastname: Wang
authors.forename: Gang
authors.initials: G
authors.lastname: Luo
authors.forename: Dian-Chun
authors.initials: DC
authors.lastname: Fang
authors.forename: Chuan
authors.initials: C
authors.lastname: Li
authors.forename: Shi-Ming
authors.initials: SM
authors.lastname: Yang
mesh_terms: D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000201:enzymology / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D002453:Cell Cycle; D049109:Cell Proliferation; D015536:Down-Regulation; D005434:Flow Cytometry; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D015316:Genetic Therapy / Q000379:methods*; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism*; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000201:enzymology / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D009361:Neoplasm Invasiveness; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction; D013997:Time Factors; D014161:Transduction, Genetic; D014162:Transfection; D023041:Xenograft Model Antitumor Assays; D000099175:Heparanase
chemical_list: D005966:Glucuronidase; D000099175:Heparanase