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<p><b>OBJECTIVE</b>To study the effects of hypoxia and hyperoxia on the expression and activity regulation of matrix metalloproteinase-2 (MMP-2) of the hepatic stellate cell (HSC).</p><p><b>METHODS</b>The expression of MMP-2, tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) and membrane type matrix metalloproteinase-1 (MT1-MMP) in cultured rat HSC under hypoxic or hyperoxic conditions were detected with immunocytochemistry (LSAB method), the contents of MMP-2, TIMP-2 in culture supernatant with ELISA and the activity of MMP-2 in supernatant with zymography.</p><p><b>RESULTS</b>(1) In the situation of hypoxia for 12 h, the expression of MMP-2 increased (hypoxia group positive indexes: 5.7 +/- 2.0; control: 3.2 +/- 1.0; P < 0.01), while TIMP-2 decreased (hypoxia group positive indexes: 2.5 +/- 0.7; control: 3.6 +/- 1.0; P < 0.05) in HSC, and the activity of MMP-2 in supernatant declined obviously (hypoxia group: 7.334 +/- 1.922; control: 17.277 +/- 7.424; P < 0.01). At the different time courses of hypoxia, the change of expression and activity of MMP-2 was most notable at 6 h. (2) In the situation of hyperoxia for 12 h, the protein contents of MMP-2, TIMP-2 in supernatant were both higher than those of the control, especially the TIMP-2 (hyperoxia group A(450): 0.050 +/- 0.014; control: 0.022 +/- 0.010; P < 0.01), and so was the activity of MMP-2 (hyperoxia group total A: 5.252 +/- 0.771; control: 4.304 +/- 1.083; P < 0.05). The expression of MT1-MMP was also increased.</p><p><b>CONCLUSIONS</b>The HSC is sensitive to the oxygen. Hypoxia accelerates the expression of MMP-2 and the effect is more marked at the early stage. Hyperoxia increases the activity of MMP-2.</p>
Subject(s)
Animals , Male , Rats , Cell Hypoxia , Physiology , Hyperoxia , Liver , Cell Biology , Matrix Metalloproteinase 2 , Metabolism , Rats, Sprague-Dawley , Tissue Inhibitor of Metalloproteinase-2ABSTRACT
PurposeTo establish sensitive and reliable serum marker on liver fibrosis for early clinical diagnosis. MethodsSerum hyaluronate acid,type Ⅲ procollagen and type Ⅳ procogen of 48 patients with viral hepatitis were analyzed by radioimmunoassay and ELISA respectively. ResultsThe serum HA, PCⅢ and PCⅣ level increased gradually with the progress of the disease. Their levels were correlated distinctly with the index of hepatic activity of inflammation and that of fibrosis.ConclusionsThe value of serum HA,PCⅢ and PCⅣ in predicting liver fibrosis will be enhanced ff the effect of inflammation could be excluded.
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Objective To study the expression of matrix metalloproteinase (MMP) and the tissue inhibitor of matrix metalloproteinase (TIMP) in hepatocellular carcinoma (HCC) and its relation with the prognosis of patient. Methods MMP 1,MMP 2,MMP 9 and TIMP 2 in human HCC tissue and HCC cell line were measured and quantified with immunohistochemistry, Northern blot hybridization and image analysis. Results MMP 1,MMP 2 and MMP 9 were localized in the cytoplasm of tumor cells. The 5 years survival rate were markedly higher in the cases of MMP 1 negative, MMP 9 negative, both MMP 1 and MMP 2 negative or both MMP 1 and MMP 9 negative staining than that of the positive cases ( P
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Objective To explore the expression and significance of p21WAF1 and p53 in HCC. Methods Immunohistochemical method (IHC) was used to localize and semi-quantitate the proteins of p21WAF1 and p53 and to observe the relationship between the expression of p21WAF1 and the different histopathologic characters in 38 patients of HCC and their peri-cancer tissue as well as 5 normal liver tissue. Results Of all 38 cases, both p21WAF1 and p53 expression were significantly higher in tumor than that in corresponding non-tumors liver tissue; 14 (36.8 %) of 38 cases showed p21WAF1 positive staining, 28 cases (73.7 %) were p53 positive, p21WAF1＋/p53＋ or p21WAF1-/p53－ were observed in 18, while 20 cases showed p53＋/p21WAF1－ or p53－/p21WAF1＋. p21WAF1＋ was seen in 1 of 38 (2.6 %) corresponding non-cancerous tissue and 2 of 5 normal liver tissue. p53 protein was not detected neither in the non-tumorous tissue nor in normal liver. No significant association was found between the expressions of p21WAF1 and p53(P＞0.05) in HCC. Their was no significant correlation between p21WAF1 or p53 expression and the different histopathologic characters of tumor (differentiating grades, intrahepatic metastasis and/or cancerous thrombi within portal veins). Conclusion Both p21WAF1 and p53 proteins are over expressed in HCC than that in corresponding non-tumorous liver tissue, but there is no relationship between them. Both p53-independent and p53-dependent mechanism may play a role in regulating p21WAF1 expression in HCC. p21WAF1 immunostaining cannot be used to assess the status of p53 in any given cell or tissue.
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Purpose:To investigate the effects of tumor suppressor gene p53 on the activity of telomerase/hTERT in human HCC-9204 cell line.Methods:Lipofection-mediated gene transfection method was used to transfect wild-type p53(wt-p53) gene into HCC 9204 cell which expresses telomerse /hTERT and carries mutant-type p53 gene. The expression of p53 was confirmed by Western blot. Both the telomerase activity and hTERT mRNA expression were detected by PCR-ELISA,TRAP-silver staining,in-situ hybridization and RT-PCR methods. The apoptotic appearance was examined by FCM.Results:Higher telomerase activity and hTERT mRNA level were detected in HCC-9204,and they were markedly inhibited after transfection with wt-p53. Meanwhile,decreasing level of bcl-2 protein and appearance of apoptosis were also shown in the transfected cells.Conclusions:Over expression of the exogenous wt-p53 gene does suppress both telomrease activity and hTERT mRNA expression in HCC cell line. There is a p53-dependent regulatory pathway for activation and expression of telomerase/hTERT in HCC.