ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of antisense cDNA of cyclin D1 on the cyclin D1 gene expression and cell proliferation of human hepatocarcinoma HepG2 cells in vitro.</p><p><b>METHODS</b>Plasmids containing cyclin D1 antisense cDNA were constructed and transfected into HepG2 cells. Their effects on cell proliferation were examined by MTT method, RT-PCR, immunohistochemical means, and flow cytometry.</p><p><b>RESULTS</b>Cyclin D1 antisense cDNA significantly inhibited the growth of HepG2 cells. The inhibition peaked at 48 hour after transfection by MTT method. RT-PCR analysis showed that cyclin D1 antisense cDNA down-regulated cyclin D1 at the mRNA levels. Expression level of cyclin D1 protein was also decreased as shown by immunohistochemical studies. Cell-cycle analysis by flow cytometry showed that transfected HepG2 cells were arrested at the G1 phase of the cell cycle.</p><p><b>CONCLUSIONS</b>Our data suggest that cyclin D1 antisense cDNA could specifically inhibit the expression of cyclin D1 mRNA and protein and regulate cell cycle and cell proliferation of HepG2 cells. Cyclin D1 antisense cDNA may serve as a potential antitumor strategy in regulating cell-cyclin treating advanced HCCs.</p>
Subject(s)
Humans , Carcinoma, Hepatocellular , Metabolism , Pathology , Cell Line, Tumor , Cyclin D1 , Genetics , Genes, bcl-1 , Genetics , Liver Neoplasms , Metabolism , Pathology , Oligodeoxyribonucleotides, Antisense , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To study the therapeutic potential of TRAIL and in combination with subtoxic level of chemotherapeutic agents in the treatment of human hepatocellular carcinomas (HCCs).</p><p><b>METHODS</b>The plasmid pcDNA 3.0-hTRAIL was transfected into COS-7 cells, and it was transiently expressed. The cytotoxic functions of the expressed product and in combination with chemotherapeutic agents were detected.</p><p><b>RESULTS</b>The transiently transfected COS-7 could express the active human TRAIL. It had mild cytotoxic functions on HCC cell lines. The cytotoxicity rates in both cell lines were 9.2% and 9.5% respectively. Chemotherapeutic agents, mitomycin (M), 5-FU and actinomycin D (AcD) dramatically augmented TRAIL induced cytotoxic function. There were significant differences in the cytotoxicity between the use of single agent and in combination with TRAIL (TRAIL + M, M; 60.1%, 32.6%, TRAIL + 5-FU, 5-FU: 68.1%, 29.2%, TRAIL + AcD, AcD; 72.9%, 58.6%) (P < 0.05).</p><p><b>CONCLUSIONS</b>The combination of human TRAIL and chemotherapeutic agents, such as mitomycin, 5-FU and actinomycin D, seemed to exert a synergistic effect compared with either agent alone, and it might have therapeutic potential in the treatment of human HCC.</p>
Subject(s)
Animals , Humans , Apoptosis Regulatory Proteins , COS Cells , Carcinoma, Hepatocellular , Drug Therapy , Pathology , Cell Line, Tumor , Liver Neoplasms , Drug Therapy , Pathology , Membrane Glycoproteins , Pharmacology , Therapeutic Uses , Recombinant Proteins , Therapeutic Uses , TNF-Related Apoptosis-Inducing Ligand , Tumor Necrosis Factor-alpha , Pharmacology , Therapeutic UsesABSTRACT
<p><b>AIM</b>To investigate the anti-inflammatory mechanism of esculentoside A (EsA) and to observe the effects of EsA on cellular adhesion between human umbilical vein endothelial cell (VEC304) and human neutrophil and to further observe the mRNA expression of intercellular adhesion molecule-1 (ICAM-1) and cluster of differentiation 18(CD18).</p><p><b>METHODS</b>The hemocyte counting method was used for assaying the adhesion rate between VEC304 and neutrophil. The RT-PCR method was used for measuring the mRNA expression of ICAM-1 and CD18.</p><p><b>RESULTS</b>The adhesion rate between VEC304 and neutrophil was increased with treatment of lipopolysaccharide(LPS). EsA (3 - 12 x 10(-6) mumol.L-1) was shown to inhibit the high cellular adhesion induced by LPS. A further investigation of adhesion molecules mRNA expression was undertaken using semi-quantitative reverse transcribed polymerase chain reaction (RT-PCR). The results of RT-PCR from VEC304 and human neutrophil treating with LPS showed that ICAM-1 and CD18 mRNA expressions were higher than those of normal cells, while this increased expression of ICAM-1 and CD18 mRNA was remarkably attenuated by the addition of EsA.</p><p><b>CONCLUSION</b>EsA was found to inhibit the increased adhesion rate induced by LPS. Moreover, LPS induced high expression of ICAM-1 and CD18 was inhibited with treatment of EsA. It might be involved in the mechanisms of anti-inflammation of EsA.</p>
Subject(s)
Adult , Humans , CD18 Antigens , Genetics , Cell Adhesion , Cell Line , Drugs, Chinese Herbal , Pharmacology , Endothelial Cells , Metabolism , Physiology , Intercellular Adhesion Molecule-1 , Genetics , Neutrophils , Metabolism , Physiology , Oleanolic Acid , Pharmacology , Phytolacca , Chemistry , Plants, Medicinal , Chemistry , RNA, Messenger , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Saponins , Pharmacology , Umbilical Veins , Cell BiologyABSTRACT
Objective: To investigate the influence of esculentoside A（EsA） on production of IL-1 and TNF by rabbit synovial cells induced by LPS. Methods: levels of IL-1 and TNF in the supernatant of rabbit synovial cell were determined by examining proliferation of thymic cells and by bioassay L929 cells as target cells, respectively. Results: EsA in 5-40 μg/ml could significantly inhibit the production of IL-1 and TNF from rabbit synovial cells induced by LPS. Conclusion: EsA can inhibit the production of IL-1 and TNF from synovial cells. It suggests that EsA may play a role in improving the rheumatoid arthritis.
ABSTRACT
Objective: To investigate the influence of esculentoside A（EsA） on production of IL-1 and TNF by rabbit synovial cells induced by LPS. Methods: levels of IL-1 and TNF in the supernatant of rabbit synovial cell were determined by examining proliferation of thymic cells and by bioassay L929 cells as target cells, respectively. Results: EsA in 5-40 μg/ml could significantly inhibit the production of IL-1 and TNF from rabbit synovial cells induced by LPS. Conclusion: EsA can inhibit the production of IL-1 and TNF from synovial cells. It suggests that EsA may play a role in improving the rheumatoid arthritis.