lipofectamine and Tongue-Neoplasms

Angiopoietin-2 (Ang-2) has been identified as an important factor in tumour angiogenesis through its action in blocking the stabilizing actions of Ang-2 and leading to new tumour vessel growth in the presence of vascular endothelial growth factor (VEGF). In the authors' previous study, over-expression of Ang-2 in Tca8113 tongue tumour cells inhibited growth. The current study aims to clarify the mechanisms of Ang-2-mediated tumour growth inhibition and its role in the regulation of VEGF expression. These studies used tumours derived from Ang-2-transfected Tca8113 cells injected into nude mice. The results showed that Ang-2-transfected tumours displayed aberrant angiogenic vessels with few associated smooth muscle cells. No detectable differences in VEGF expression were observed between Ang-2-transfected and parental tumours. Tumours produced by the Ang-2 transfection also had a higher apoptosis index and lower tumour cell proliferative index than tumours in the control groups. These observations suggest that enhanced expression of Ang-2 has no effect on VEGF expression and results in tumour vessel regression and inhibition of tumour growth.

Topics: Actins; Angiopoietin-2; Animals; Apoptosis; Carcinoma; Cell Line, Tumor; Cell Proliferation; Endothelial Cells; Endothelium, Vascular; Gene Expression Regulation, Neoplastic; Immunohistochemistry; In Situ Nick-End Labeling; Lipids; Liposomes; Mice; Mice, Nude; Microvessels; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Neoplasm Transplantation; Neovascularization, Pathologic; Plasmids; Proliferating Cell Nuclear Antigen; Reverse Transcriptase Polymerase Chain Reaction; Tongue Neoplasms; Transfection; Vascular Endothelial Growth Factor A

This study used RNA interference (RNAi) to explore the effect of NO and inducible nitric oxide synthase (iNOS) on apoptosis and proliferation in the tongue squamous carcinoma cell line Tca8113. Tca8113 cells were transfected with the plasmid pGenesil-1, which expresses iNOS short hairpin RNA (shRNA), or the negative control plasmid pSilencer-HK, and the transfected cells were compared with untransfected cells. The expression of iNOS was detected by histochemistry, and apoptosis was detected by flow cytometry. The expression of iNOS was significantly lower in the pSilencer-iNOS group than in the pSilencer-HK and empty control groups. The apoptosis rate was significantly higher in the pSilencer-iNOS group than in the pSilencer-HK and empty control groups. Growth monitoring showed that proliferation was also inhibited in cells transfected with pSilencer-iNOS. RNAi gene silencing decreased iNOS gene expression, induced apoptosis, and suppressed proliferation in Tca8113 cells.

Topics: Apoptosis; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Coloring Agents; Flow Cytometry; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Gene Silencing; Histocytochemistry; Humans; Indicators and Reagents; Lipids; Nitric Oxide; Nitric Oxide Synthase Type II; Plasmids; RNA Interference; RNA, Small Interfering; Tetrazolium Salts; Thiazoles; Tongue Neoplasms; Transfection

The purpose of this study was to establish transfergeneic Tca8113 cell and evaluate the expression of human endostatin (hES) gene in the cell colone in vitro.. To transfect hES gene into Tca8113 cells, lipofectamin was complexed with plasmid encoding hES gene, and blasticidin S antibiotic was adopted to select Tca8113--hES cell clone. Immunohistochemistry S-P method was adopted to detect the expression of hES in the transfergenic Tca8113 cell in vitro.. Transfected by hES, the transfergenic Tca8113 cells could grow and proliferate in RPMI--1640 culture medium containing blasticidin S antibiotic. The expression rate of hES reached 100%.. hES gene can express in hES-transfected Tca8113 cell in vitro.

Topics: Carcinoma, Squamous Cell; Cell Division; Cloning, Molecular; Endostatins; Humans; Lipids; Tongue Neoplasms; Transfection; Tumor Cells, Cultured