ABSTRACT
Previous researches showed that the expression level of E-Cad in most infiltrating cancer cells was reduced or negative.This study explored whether 4HPR restrained the infiltration of bladder cancer cells through regulating the expression of E-Cad.The infiltrating bladder cancer cells T24 were cultured,and then treated by a proper dosage of drug.Their viability was a determined by MTT method.Western blotting and RT-PCR were adopted to detect the changes of E-Cad gene expression at both protein and mRNA levels.Moreover,immunofluorescent staining and confocal fluorescence microscopy were employed for the observation of the expression of E-Cad.The result showed that,at both mRNA and protein levels,the expression level of E-Cad in T24 cells treated by 4HPR was significantly higher than that of control group,while the β-Cat expression was also relocated from the cell nucleus to cytoplasm.Our findings suggested that the regulatory function of 4HPR on infiltration of bladder cancer cells T24 is at least partly achieved by regulating the expression of E-Cad.
ABSTRACT
The action mechanism of matrix metalloproteinases-2 (MMP-2) and tissue inhibitor of metalloproteinases-2 (TIMP-2) in the genesis, development and degeneration of haemangioma was in-vestigated by detecting their expression in the tissue of haemangioma in different phases by using the immunohistochemistry. Fifty paraffin-embedded specimens of skin capillary haemangioma were col-lected, which were documented in the Department of Pathology, Renmin Hospital of Wuhan University from 2000 to 2006. All samples were stained by regular HE method, and proliferative cell nuclear anti-gen (PCNA) was tested by immunohistochemical S-P method, The samples were classified according to the Mulliken criteria and the expression pattern of PCNA. Immunohistochemical S-P method was ap-plied to detect the expression of MMP-2 and TIMP-2 in proliferative and degenerative phases of cuta-neous capillary haemangioma, and in normal skin tissues. In combination with the detection of the ex-pression of factor Ⅷ-related antigen, it was verified that in haemangioma tissues, the cells expressing MMP-2 and TIMP-2 were vascular endothelial cells. The MMP-2 and TIMP-2 expression was quantita-tively analyzed by image analysis system (HPIAS-1000), and one-way ANOVA(107) and SNK(q) test were done to analyze average absorbance (A) and positive area rate of immunohistochemically positive particles by using SPSS11.5. The results showed: (1) Among 50 samples of haemangioma, there were 26 proliferative haemangiomas, and 24 degenerative haemangiomas, respectively; (2) The expression of MMP-2 was weak in normal vascular endothelial cells, cytoplasm of connective tissues and extracellu-lar matrix around blood vessels. The expression of MMP-2 in proliferative group was significantly higher than in degenerative group and control group (normal skin) (P<0.05), but there was no statisti-cally significant difference between the latter two groups; (3) TIMP-2 was highly expressed in normal tissues, degenerative vascular endothelial cells, cytoplasm of connective tissues and extracellular matrix around blood vessels. The expression level of TIMP-2 in proliferative phase was significantly lower than in degenerative phase (P<0.05), and the expression of TIMP-2 in proliferative phase was signifi-cantly different from that in degenerative phase and normal tissues (P<0.05). It was concluded that in proliferative phase of haemangioma, MMP-2 may promote over-proliferation of endothelial cells of haemangioma, and in degenerative phase, TIMP-2 can inhibit the proliferation of endothelial cells of haemangioma. The two substances play important roles in the genesis, development and degeneration of haemangiomas.