ABSTRACT
<p><b>BACKGROUND</b>Esophageal cancer is the sixth leading cause of cancer-related death worldwide. Pentraxin-3 (PTX3) is a member of the PTX superfamily. Here, we investigated the role of PTX3 in esophageal squamous cell carcinoma (ESCC).</p><p><b>METHODS</b>The effect of PTX3 on ESCC cell proliferation, colony formation, apoptosis, migration, and invasion was investigated using cell viability assays, colony formation assays, flow cytometry, and migration and invasion assays. The effect of PTX3 on the tumorigenicity of ESCC in vivo was investigated with xenograft studies in nude mice.</p><p><b>RESULTS</b>PTX3 overexpression in ESCC cells reduced cellular proliferation and colony formation (P < 0.05) and increased the rate of apoptosis (P < 0.05). PTX3 expression had no significant effect on the migratory or invasive potential of ESCC cells. In our mouse model of human ESCC, we achieved 100% successful tumor establishment. Compared with the control and empty vector-expressing groups, the PTX3-expressing group formed significantly smaller tumors (P < 0.05).</p><p><b>CONCLUSIONS</b>This study indicates that PTX3 might play an inhibitory role in ESCC.</p>
Subject(s)
Animals , Humans , Male , Mice , Apoptosis , Genetics , Physiology , C-Reactive Protein , Genetics , Metabolism , Carcinoma, Squamous Cell , Metabolism , Pathology , Cell Line, Tumor , Cell Proliferation , Genetics , Physiology , Cell Survival , Genetics , Physiology , Esophageal Neoplasms , Metabolism , Pathology , Mice, Inbred BALB C , Mice, Nude , Serum Amyloid P-Component , Genetics , Metabolism , Xenograft Model Antitumor AssaysABSTRACT
<p><b>BACKGROUND</b>Hepatitis B is at particularly high risk of fibrosis progression. Unfortunately, the mechanism of hepatic fibrogenesis induced by hepatitis B virus (HBV) has not been fully understood to date. The aim of this study was to observe whether HBV can infect hepatic stellate cells (HSCs), and to examine the effects of HBV or HBV S protein (HBs) on the proliferation and collagen type I expression of HSCs.</p><p><b>METHODS</b>The supernatants of HepG2.2.15 cells which contained HBV-DNA or HBs were added to LX-2 cells for 72 hours. Cell survival was determined by MTT assay. HBV particles in LX-2 cells were detected by transmission electron microscopy. The expression of HBs and HBV C protein (HBc) was determined by confocal fluorescence microscopy. The expression levels of HBV-DNA were measured by real-time PCR. The cellular collagen type I mRNA and protein levels were quantified by reverse transcription-PCR and ELISA, respectively.</p><p><b>RESULTS</b>High concentrations of HBV (1.2 x 10(5) - 5.0 x 10(5) copies/ml) or HBs (1.25 - 20 microg/ml) inhibited the proliferation of LX-2 cells, while low concentrations of HBV (1.0 x 10(3) - 6.2 x 10(4) copies/ml) or HBs (0.04 - 0.62 microg/ml) promoted the proliferation. After treating LX-2 cells with HBV for 72 hours, about 42 nm HBV-sized particles and strong expression of HBs and HBc were found in the cytoplasm of LX-2 cells. HBV-DNA in the culture medium of LX-2 cells decreased at 24 hours, rose at 48 hours and thereafter, decreased again at 72 hours. The mRNA and protein expression of cellular collagen type I in LX-2 cells were significantly increased by HBV infection but not by recombinant HBs.</p><p><b>CONCLUSIONS</b>HBV and HBs affect the proliferation of HSCs; HBV can transiently infect and replicate in cultured HSCs and express HBs and HBc in vitro. Furthermore, HBV can significantly increase the expression of collagen type I mRNA and protein in HSCs.</p>
Subject(s)
Humans , Cell Line , Cell Proliferation , Collagen Type I , Metabolism , Enzyme-Linked Immunosorbent Assay , Gene Expression Regulation , Hepatic Stellate Cells , Metabolism , Virology , Hepatitis B virus , Physiology , Microscopy, Confocal , Microscopy, Electron, Transmission , Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To express and purify the recombinant human pigment epithelium-derived factor (PEDF) which inhibits the proliferation of the endothelium cells from blood vessel in E.coli.</p><p><b>METHODS</b>PEDF gene was inserted into the prokaryotic expression vector pGEX-4T-2. The recombinant protein PEDF was expressed in E.coli BL-21, and purified by the GST Sepharose 4B affinity column. The recombinant human PEDF protein was identified by Western blot and mass spectrum. The biological activity of the recombinant human PEDF protein was measured by using MTT.</p><p><b>RESULTS</b>The 46 kDa recombinant human PEDF protein was obtained. It significantly inhibited the proliferation of the human umbilical vein cell line HUVEC.</p><p><b>CONCLUSION</b>The recombinant human PEDF with anti-angiogenesis activity protein may be successfully purify through prokaryotic expression.</p>