Objective： To study the expression of extracellular matrix （ECM） proteins including Collagen Ⅳ （Co Ⅳ）, Fibronectin, Laminin in human non-small cell lung cancer （NSCLC） specimens and the relationship between E...Objective： To study the expression of extracellular matrix （ECM） proteins including Collagen Ⅳ （Co Ⅳ）, Fibronectin, Laminin in human non-small cell lung cancer （NSCLC） specimens and the relationship between ECM and cell adhesion, proliferation, apoptosis and drug sensitivity in NSCLC cell line. And to investigate the role of phosphatidylinositol 3-kinase （PI3-K） in signal transduction of Co Ⅳ in NSCLC. Methods： The expression of ECM proteins was detected by using immunohistochemical staining （Envision＇s）. Adherent cells were stained with 1% methylene blue. Cell proliferation and cytotoxic effects were monitored by MTT assay. Cell apoptosis was analyzed by FITC-Annexin V/PI double staining variables flow cytometry （FCM）. Results： The expression rate of Co Ⅳ （93%） was the highest compared to others in NSCLC stroma. After treated with Co Ⅳ, the adhesion of H1299 cells was increased and the cytotoxicity of cis-platinum （DDP） against H1299 cells was decreased compared to the control （P〈0.05）. After treated with Co Ⅳ both survival and proliferation rates were higher and apoptosis rate was lower than without Co Ⅳ （P〈0.05）. PI3-K inhibitor LY294002 decreased both survival and proliferation rates （82.7%±2.0% and 75.2%±6.8%, respectively）, even on Co Ⅳ-coated surface （92.2%±2.8% and 84.6%±9.2%, respectively）. And it also helped DDP increase apoptosis. Conclusion： ECM remodeling existed in NSCLC. Co Ⅳ protected NSCLC cells from DDP-induced apoptosis and weakened the cytotoxicity of DDP. PI3-K pathway might be the crucial mechanism of apoptosis impairment and drug resistance.展开更多
This study was conducted to investigate the effect of dietary proline(Pro), and Pro and hydroxyproline(Hyp) in combination on the growth performance, total Hyp and collagen concentrations of tissues, and prolyl 4-hydr...This study was conducted to investigate the effect of dietary proline(Pro), and Pro and hydroxyproline(Hyp) in combination on the growth performance, total Hyp and collagen concentrations of tissues, and prolyl 4-hydroxylase α(I)(P4H α(I)) gene expression in juvenile turbot feeding high plant protein diets. A diet containing 50% crude protein and 12% crude lipid was formulated as the basal and control, on which other two protein and lipid contents identical experimental diets were formulated by supplementing the basal with either 0.75% Pro(Pro-0.75) or 0.75% Pro and 0.75% Hyp(Pro+Hyp). Four groups of fish in indoor seawater recirculating systems, 35 individuals each, were fed twice a day to apparent satiation for 10 weeks. The results showed that dietary Pro and Hyp supplementation had no significant effect on growth performance and feed utilization of juvenile turbot(P > 0.05). Total Hyp and collagen concentrations in muscle were significantly increased when dietary Pro and Hyp increased(P < 0.05), and fish fed diet Pro+Hyp showed significantly higher free Hyp content in plasma than those fed other diets(P < 0.05). The expression of P4 H α(I) gene in liver and muscle was significantly up regulated in fish fed diet Pro-0.75 in comparison with control(P < 0.05); however the gene was significantly down regulated in fish fed diet Pro+Hyp in muscle in comparison with fish fed diet Pro-0.75(P < 0.05). It can be concluded that supplement of crystal L-Pro and L-Hyp to high plant protein diets did not show positive effects on growth performance of juvenile turbot, but enhanced total collagen concentrations in muscle.展开更多
Objective: To study the effects of quercetin (QU) on matrix metallo-proteinases (MMPs) , the tissue inhibitor of matalloproteinase-1 (TIMP-1) , procollagen I and 2 proteoglycans (decorin and biglycan) mRNA expression ...Objective: To study the effects of quercetin (QU) on matrix metallo-proteinases (MMPs) , the tissue inhibitor of matalloproteinase-1 (TIMP-1) , procollagen I and 2 proteoglycans (decorin and biglycan) mRNA expression in cultured rat hepatic stellate cell line HSC-T6 cells. Methods: Cells were treated with different concentrations of QU (12. 5, 25, 50 μmol/L) or drug solvent (0. 1 % Me2SO) for 24 h. mRNA expression was determined by reverse transcription polymerase chain reaction (RT-PCR). Results: QU (12.5 - 50 μmol/L) enhanced collagenase (rat MMP-13) and membrane typel-MMP (MMP-14) mRNA expression, decreased procollagen I mRNA expression in a concentration-dependent manner, but did not affect gelatinase-A (MMP-2) , TIMP-1, decorin and biglycan expression. Conclusion: QU may decrease matrix deposition and increase matrix degradation, which might be beneficial to liver fibrosis.展开更多
Objective:To study gene expression of collagen types IX and X in human lumbar intervertebral discs during aging and degeneratio n and to explore the role of collagen types IX and X in disc degeneration. Methods:Fetal,...Objective:To study gene expression of collagen types IX and X in human lumbar intervertebral discs during aging and degeneratio n and to explore the role of collagen types IX and X in disc degeneration. Methods:Fetal, adult and pathologic specimens were subjected t o in situ hybridization with cDNA probes to investigate mRNA-expressions of typ es IX and X collagen gene. Results:In fetal intervertebral discs, positive mRNA hybridiza tion signals of type IX collagen were concentrated in the nucleus pulposus and t he inner layer of anulus fibrosus. Interstitial matrix of the nucleus pulposus a lso showed positive type X collagen staining. Positive mRNA hybridization signal s of types IX and X were not detected in the middle and outer layers of anulus f ibrosus. In adult specimens, expression of type IX collagen mRNA was markedly de creased. No hybridization signals of type X collagen was observed. As for pathol ogical specimens, there was no gene expression of type IX collagen. In severe de generated discs from adults, there were focal positive expressions of type X col lagen. Conclusions:Obvious changes of collagen gene expression occur with aging. Expression of type IX collagen decreases in adult and pathological d iscs. Results of type X collagen expression suggest that type X collagen is expr essed only in older adult and senile discs (i.e., when disc degeneration has alr eady reached a terminal stage), indicating the terminal stage of degeneration.展开更多
To analyze the expression of procollagen gene in fracture callus, and to search for the technique of in situ hybridization for undecalcified skeletal tissue. Methods: In situ hybridization of procollagen gene expres...To analyze the expression of procollagen gene in fracture callus, and to search for the technique of in situ hybridization for undecalcified skeletal tissue. Methods: In situ hybridization of procollagen gene expression was performed on the undecalcified cryosections of rat fracture callus at 7, 14, and 28 d. Results: The hybridization signals achieved were clear and easy to be localized with high specificity. On the 7th day, the expressions of pro α1(Ⅲ) in fibroblasts and some chondrocyte like cells were dominant; and at the end of second week high expression of type Ⅱ procollagen mRNA was observed in chondrocytes. At the end of fourth week, the cartilaginous callus was almost all replaced by woven bone tissue, and some type Ⅰ procollagen mRNA positive osteoblasts and hypertrophic chondrocytes were found scattering in the woven bone and remnants of cartilaginous callus.Conclusions: The modified method employed in this study is easier, quicker, and more sensitive with high specificity than the conventional technique for in situ hybridization of procollagen gene expression of decalcified rat fracture callus. The phenomenon of shared phenotype expression, which was demonstrated among cells engaged in fracture healing, indicates an important approach to reveal the mechanism of the origin, differentiation, and orientation of cells.展开更多
基金This project was supported by the Science Foundation of Shanghai Municipal Commission of Science and Technology (034119953).
文摘Objective： To study the expression of extracellular matrix （ECM） proteins including Collagen Ⅳ （Co Ⅳ）, Fibronectin, Laminin in human non-small cell lung cancer （NSCLC） specimens and the relationship between ECM and cell adhesion, proliferation, apoptosis and drug sensitivity in NSCLC cell line. And to investigate the role of phosphatidylinositol 3-kinase （PI3-K） in signal transduction of Co Ⅳ in NSCLC. Methods： The expression of ECM proteins was detected by using immunohistochemical staining （Envision＇s）. Adherent cells were stained with 1% methylene blue. Cell proliferation and cytotoxic effects were monitored by MTT assay. Cell apoptosis was analyzed by FITC-Annexin V/PI double staining variables flow cytometry （FCM）. Results： The expression rate of Co Ⅳ （93%） was the highest compared to others in NSCLC stroma. After treated with Co Ⅳ, the adhesion of H1299 cells was increased and the cytotoxicity of cis-platinum （DDP） against H1299 cells was decreased compared to the control （P〈0.05）. After treated with Co Ⅳ both survival and proliferation rates were higher and apoptosis rate was lower than without Co Ⅳ （P〈0.05）. PI3-K inhibitor LY294002 decreased both survival and proliferation rates （82.7%±2.0% and 75.2%±6.8%, respectively）, even on Co Ⅳ-coated surface （92.2%±2.8% and 84.6%±9.2%, respectively）. And it also helped DDP increase apoptosis. Conclusion： ECM remodeling existed in NSCLC. Co Ⅳ protected NSCLC cells from DDP-induced apoptosis and weakened the cytotoxicity of DDP. PI3-K pathway might be the crucial mechanism of apoptosis impairment and drug resistance.
基金financially supported by the China Agriculture Research System (CARS-50-G08)Agricultural Scientific and Technological Achievements into Capital (2010GB23600673)the National Natural Science Foundation of China (Grant Nos. 31072222 and 30901 108)
文摘This study was conducted to investigate the effect of dietary proline(Pro), and Pro and hydroxyproline(Hyp) in combination on the growth performance, total Hyp and collagen concentrations of tissues, and prolyl 4-hydroxylase α(I)(P4H α(I)) gene expression in juvenile turbot feeding high plant protein diets. A diet containing 50% crude protein and 12% crude lipid was formulated as the basal and control, on which other two protein and lipid contents identical experimental diets were formulated by supplementing the basal with either 0.75% Pro(Pro-0.75) or 0.75% Pro and 0.75% Hyp(Pro+Hyp). Four groups of fish in indoor seawater recirculating systems, 35 individuals each, were fed twice a day to apparent satiation for 10 weeks. The results showed that dietary Pro and Hyp supplementation had no significant effect on growth performance and feed utilization of juvenile turbot(P > 0.05). Total Hyp and collagen concentrations in muscle were significantly increased when dietary Pro and Hyp increased(P < 0.05), and fish fed diet Pro+Hyp showed significantly higher free Hyp content in plasma than those fed other diets(P < 0.05). The expression of P4 H α(I) gene in liver and muscle was significantly up regulated in fish fed diet Pro-0.75 in comparison with control(P < 0.05); however the gene was significantly down regulated in fish fed diet Pro+Hyp in muscle in comparison with fish fed diet Pro-0.75(P < 0.05). It can be concluded that supplement of crystal L-Pro and L-Hyp to high plant protein diets did not show positive effects on growth performance of juvenile turbot, but enhanced total collagen concentrations in muscle.
文摘Objective: To study the effects of quercetin (QU) on matrix metallo-proteinases (MMPs) , the tissue inhibitor of matalloproteinase-1 (TIMP-1) , procollagen I and 2 proteoglycans (decorin and biglycan) mRNA expression in cultured rat hepatic stellate cell line HSC-T6 cells. Methods: Cells were treated with different concentrations of QU (12. 5, 25, 50 μmol/L) or drug solvent (0. 1 % Me2SO) for 24 h. mRNA expression was determined by reverse transcription polymerase chain reaction (RT-PCR). Results: QU (12.5 - 50 μmol/L) enhanced collagenase (rat MMP-13) and membrane typel-MMP (MMP-14) mRNA expression, decreased procollagen I mRNA expression in a concentration-dependent manner, but did not affect gelatinase-A (MMP-2) , TIMP-1, decorin and biglycan expression. Conclusion: QU may decrease matrix deposition and increase matrix degradation, which might be beneficial to liver fibrosis.
文摘Objective:To study gene expression of collagen types IX and X in human lumbar intervertebral discs during aging and degeneratio n and to explore the role of collagen types IX and X in disc degeneration. Methods:Fetal, adult and pathologic specimens were subjected t o in situ hybridization with cDNA probes to investigate mRNA-expressions of typ es IX and X collagen gene. Results:In fetal intervertebral discs, positive mRNA hybridiza tion signals of type IX collagen were concentrated in the nucleus pulposus and t he inner layer of anulus fibrosus. Interstitial matrix of the nucleus pulposus a lso showed positive type X collagen staining. Positive mRNA hybridization signal s of types IX and X were not detected in the middle and outer layers of anulus f ibrosus. In adult specimens, expression of type IX collagen mRNA was markedly de creased. No hybridization signals of type X collagen was observed. As for pathol ogical specimens, there was no gene expression of type IX collagen. In severe de generated discs from adults, there were focal positive expressions of type X col lagen. Conclusions:Obvious changes of collagen gene expression occur with aging. Expression of type IX collagen decreases in adult and pathological d iscs. Results of type X collagen expression suggest that type X collagen is expr essed only in older adult and senile discs (i.e., when disc degeneration has alr eady reached a terminal stage), indicating the terminal stage of degeneration.
文摘To analyze the expression of procollagen gene in fracture callus, and to search for the technique of in situ hybridization for undecalcified skeletal tissue. Methods: In situ hybridization of procollagen gene expression was performed on the undecalcified cryosections of rat fracture callus at 7, 14, and 28 d. Results: The hybridization signals achieved were clear and easy to be localized with high specificity. On the 7th day, the expressions of pro α1(Ⅲ) in fibroblasts and some chondrocyte like cells were dominant; and at the end of second week high expression of type Ⅱ procollagen mRNA was observed in chondrocytes. At the end of fourth week, the cartilaginous callus was almost all replaced by woven bone tissue, and some type Ⅰ procollagen mRNA positive osteoblasts and hypertrophic chondrocytes were found scattering in the woven bone and remnants of cartilaginous callus.Conclusions: The modified method employed in this study is easier, quicker, and more sensitive with high specificity than the conventional technique for in situ hybridization of procollagen gene expression of decalcified rat fracture callus. The phenomenon of shared phenotype expression, which was demonstrated among cells engaged in fracture healing, indicates an important approach to reveal the mechanism of the origin, differentiation, and orientation of cells.
