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1.
Biomed J ; 39(4): 261-271, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27793268

ABSTRACT

BACKGROUND: In vitro and clinical studies suggest that the oncogene LMP1 (latent membrane protein 1) encoded by Epstein-Barr virus (EBV) plays a role in the development of nasopharyngeal carcinoma (NPC) and the formation of metastases in immunocompetent individuals. However, whether LMP1 itself is sufficient to drive these events in immunocompetent hosts remains elusive due to the lack of appropriate experimental models. The aim of this study was to study LMP1-dependent tumorigenesis and metastasis in BALB/c mice inoculated with BALB/c-3T3 cells expressing N-LMP1 (a Taiwanese NPC variant). METHODS: Following cancer cell inoculation, metastasis formation was monitored over time using PCR analysis of LMP1 as tumor marker. We also used a luciferase (Luc)-containing N-LMP1 and bioluminescent imaging (BLI) to monitor metastasis formation in a non-invasive manner. RESULTS: N-LMP1 appeared early in draining lymph nodes and in various distant organs before the rapid growth of the primary tumor. Lung metastasis was observed by BLI and further confirmed by histological examination. Furthermore, we detected luciferase signals in the lungs, even before the animals were sacrificed. CONCLUSIONS: Our results demonstrate the high metastatic character of N-LMP1 in immunocompetent hosts. Systemic tumor dissemination occurs even before aggressive tumor growth at the primary site, suggesting that early treatment of primary LMP1-associated tumors and distant micro-metastases is critical to achieve positive results.


Subject(s)
Nasopharyngeal Neoplasms/pathology , Neoplasm Metastasis/pathology , Viral Matrix Proteins/physiology , Animals , Herpesvirus 4, Human , Immunocompetence , Lung Neoplasms/secondary , Mice , Mice, Inbred BALB C , Neoplasms, Experimental/pathology
2.
Clin Oral Investig ; 16(4): 1171-80, 2012 Aug.
Article in English | MEDLINE | ID: mdl-21837388

ABSTRACT

Stem cell therapy is a promising area in regenerative medicine. Periodontal granulation tissues are often discarded during conventional surgery. If stromal stem cells can be isolated from these tissues, they can be used for subsequent surgery on the same patient. Fifteen human periodontal granulation tissue samples were obtained from intrabony defects during surgery. Immunohistochemistry (IHC) was carried out on five of the samples to identify STRO-1, a marker of mesenchymal stem cells. Five samples underwent flow cytometry analysis for the same marker. The remaining five samples were characterized by "colony formation unit-fibroblast" (CFU-f) assay and selected for proliferation assay, flow cytometry of stem cell markers, immunocytochemistry (ICC), multipotent differentiation assays, and repairing critical-size defects in mice. The ratio of STRO-1(+) cells detected by IHC was 5.91 ± 1.50%. The analysis of flow cytometry for STRO-1 was 6.70 ± 0.81%. Approximately two thirds of the CFU-f colonies had a strong reaction to STRO-1 in ICC staining. The cells were multipotent both in vitro and in vivo. Mice given bone grafts and stem cells showed significantly better bone healing than those without stem cells. Multipotent stromal stem cells can be isolated from human periodontal granulation tissues. These cells improve new bone formation when transplanted in mouse calvarial defects. Isolating stem cells from relatively accessible sites without extra procedures is clinically advantageous. This study demonstrated that human periodontal granulation tissues contain isolatable multipotent stem cells. The cells may be a good source for autotransplantation in subsequent treatment.


Subject(s)
Granulation Tissue/cytology , Mesenchymal Stem Cells/cytology , Periodontium/cytology , Adipogenesis/physiology , Alveolar Bone Loss/pathology , Alveolar Bone Loss/surgery , Animals , Antigens, Surface/analysis , Bone Diseases/surgery , Bone Substitutes/therapeutic use , Bone Transplantation/physiology , CD146 Antigen/analysis , Cell Differentiation/physiology , Cell Proliferation , Cell Separation , Chondrogenesis/physiology , Fibroblasts/cytology , Humans , Hyaluronan Receptors/analysis , Hydroxyapatites/therapeutic use , Mesenchymal Stem Cell Transplantation , Mice , Mice, Inbred NOD , Multipotent Stem Cells/cytology , Osteogenesis/physiology , Skull/surgery , Thy-1 Antigens/analysis , Tissue Scaffolds
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