title: Overexpression of RUNX3 inhibits malignant behaviour of Eca109 cells in vitro and vivo.
abstract: Runt-related transcription factor 3 (RUNX3) is a tumor suppressor gene whose reduced expression may play an important role in the development and progression of esophageal squamous cell cancer (ESCC). The aim of this study was to investigate the clinical relevance of RUNX3 in ESCC patients and effects of overexpression on biological behaviour of Eca109 cells in vitro and in vivo. Immunohistochemistry was performed to detect the clinical relevance of RUNX3 and lymph node metastasis in 80 ESCC tissues and 40 non-cancerous tissues using the SP method. RT-PCR and Western blotting were applied to assess the RUNX3 level and verify the Eca109 cell line with stable overexpression. Localization of RUNX3 proteins was performed by cell immunofluorescence. CCK-8 and Scrape motility assays were used to determine proliferation and migration and the TUNEL assay to analyze cell apoptosis. Invasive potential was assessed in cell transwell invasion experiments. In nude mice, tumorigenesis in vivo was determined. Results showed decreased expression of RUNX3 in esophageal tissue to be significantly related to lymph node metastasis (LNM) (P<0.01). In addition, construction of a recombinant lentiviral vector and transfection into the human ESCC cell line Eca109 demonstrated that overexpression could inhibit cell proliferation, migration and invasion, and induce apoptosis. The in vivo experiments in mice showed tumorigenicity and invasiveness to be significantly reduced. Taken together, our studies indicate that underexpression of RUNX3 in human ESCC tissue is significantly correlated with progression. Restoration of RUNX3 expression significantly inhibits ESCC cells proliferation, migration, invasion and tumorigenesis.
authors.affiliation: Department of Thoracic Surgery, Provincial Hospital Affiliated to Shandong University, Jinan, China E-mail :wz620226@hotmail.com.
authors.forename: Hua-Xia
authors.initials: HX
authors.lastname: Chen
authors.forename: Shuai
authors.initials: S
authors.lastname: Wang
authors.forename: Zhou
authors.initials: Z
authors.lastname: Wang
authors.forename: Zhi-Ping
authors.initials: ZP
authors.lastname: Zhang
authors.forename: Shan-Shan
authors.initials: SS
authors.lastname: Shi
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000473:pathology*; D050677:Core Binding Factor Alpha 3 Subunit / Q000096:biosynthesis* / Q000235:genetics; D018450:Disease Progression; D004938:Esophageal Neoplasms / Q000235:genetics / Q000473:pathology*; D000077277:Esophageal Squamous Cell Carcinoma; D006801:Humans; D008207:Lymphatic Metastasis / Q000235:genetics / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009361:Neoplasm Invasiveness / Q000235:genetics / Q000473:pathology*; D012333:RNA, Messenger / Q000096:biosynthesis; D012844:Sincalide / Q000096:biosynthesis; D025521:Tumor Suppressor Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D050677:Core Binding Factor Alpha 3 Subunit; D012333:RNA, Messenger; C493729:Runx3 protein, human; D025521:Tumor Suppressor Proteins; D012844:Sincalide