AAV-mediated human PEDF inhibits tumor growth and metastasis in murine colorectal peritoneal carcinomatosis modelReport as inadecuate




AAV-mediated human PEDF inhibits tumor growth and metastasis in murine colorectal peritoneal carcinomatosis model - Download this document for free, or read online. Document in PDF available to download.

BMC Cancer

, 12:129

First Online: 30 March 2012Received: 04 October 2011Accepted: 30 March 2012DOI: 10.1186-1471-2407-12-129

Cite this article as: Wu, Q.J., Gong, C.Y., Luo, S.T. et al. BMC Cancer 2012 12: 129. doi:10.1186-1471-2407-12-129

Abstract

BackgroundAngiogenesis plays an important role in tumor growth and metastasis, therefore antiangiogenic therapy was widely investigated as a promising approach for cancer therapy. Recently, pigment epithelium-derived factor PEDF has been shown to be the most potent inhibitor of angiogenesis. Adeno-associated virus AAV vectors have been intensively studied due to their wide tropisms, nonpathogenicity, and long-term transgene expression in vivo. The objective of this work was to evaluate the ability of AAV-mediated human PEDF hPEDF as a potent tumor suppressor and a potential candidate for cancer gene therapy.

MethodsRecombinant AAV2 encoding hPEDF rAAV2-hPEDF was constructed and produced, and then was assigned for in vitro and in vivo experiments. Conditioned medium from cells infected with rAAV2-hPEDF was used for cell proliferation and tube formation tests of human umbilical vein endothelial cells HUVECs. Subsequently, colorectal peritoneal carcinomatosis CRPC mouse model was established and treated with rAAV2-hPEDF. Therapeutic efficacy of rAAV2-hPEDF were investigated, including tumor growth and metastasis, survival time, microvessel density MVD and apoptosis index of tumor tissues, and hPEDF levels in serum and ascites.

ResultsrAAV2-hPEDF was successfully constructed, and transmission electron microscope TEM showed that rAAV2-hPEDF particles were non-enveloped icosahedral shape with a diameter of approximately 20 nm. rAAV2-hPEDF-infected cells expressed hPEDF protein, and the conditioned medium from infected cells inhibited proliferation and tube-formation of HUVECs in vitro. Furthermore, in CRPC mouse model, rAAV2-hPEDF significantly suppressed tumor growth and metastasis, and prolonged survival time of treated mice. Immunofluorescence studies indicated that rAAV2-hPEDF could inhibit angiogenesis and induce apoptosis in tumor tissues. Besides, hPEDF levels in serum and ascites of rAAV2-hPEDF-treated mice were significant higher than those in rAAV2-null or normal saline NS groups.

ConclusionsThus, our results suggest that rAAV2-hPEDF may be a potential candidate as an antiangiogenic therapy agent.

KeywordsAAV PEDF Tumor Antiangiogesis Gene therapy AbbreviationsANOVAOne-way analysis of variance

bFGFBasic fibroblast growth factor

CRPCColorectal peritoneal carcinomatosis

DMEMDulbecco-s Modified Eagle-s medium

ECLEnhanced chemiluminescence

ELISAEnzyme-linked immunosorbnent assay

FBSFetal bovine serum

hPEDFHuman pigment epithelium-derived factor

HUVECsHuman umbilical vein endothelial cells

HandEHematoxylin and eosin

IL-8Interleukin-8

MTTMethyl thiazolyl tetrazolium

MVDMicrovessel density

NSNormal saline

PEDFPigment epithelium-derived factor

rAAVRecombinant adeno-associated virus

rAAV2Recombinant adeno-associated virus serotype 2 vectors

rAAV2-hPEDFRecombinant adeno-associated virus serotype 2 vectors encoding human pigment epithelium-derived factor

S.D.Standard deviation

TUNELTerminal deoxynucleotidyl transferase-mediated nick-end labeling

VEGFVascular endothelial growth factor.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2407-12-129 contains supplementary material, which is available to authorized users.

Qin Jie Wu, Chang Yang Gong contributed equally to this work.

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Author: Qin Jie Wu - Chang Yang Gong - Shun Tao Luo - Dong Mei Zhang - Shuang Zhang - Hua Shan Shi - Lian Lu - Heng Xiu Yan

Source: https://link.springer.com/







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