AIM: Cyclooxygenase-2 (COX-2) has been suggested to be associated with carcinogenesis. We sought to investigate the effect of the selective COX-2 inhibitor, Nimesulide on proliferation and apoptosis of SMMC-7721 human...AIM: Cyclooxygenase-2 (COX-2) has been suggested to be associated with carcinogenesis. We sought to investigate the effect of the selective COX-2 inhibitor, Nimesulide on proliferation and apoptosis of SMMC-7721 human hepatoma cells.METHODS: This study was carried out on the culture of hepatic carcinoma SMMC-7721 cell line. Various concentrations of Nimesulide (0, 200 micromol/L, 300 micromol/L, 400 micromol/L) were added and incubated. Cell proliferation was detected with MTT colorimetric assay, cell apoptosis by electron microscopy, flow cytometry and TUNEL.RESULTS: Nimesulide could significantly inhibit SMMC-7721 cells proliferation dose-dependent and in a dependent manner compared with that of the control group. The duration lowest inhibition rate produced by Nimesulide in SMMC-7721 cells was 19.06%, the highest inhibition rate was 58.49%. After incubation with Nimesulide for 72 h, the most highest apoptosis rate and apoptosis index of SMMC-7721 cells comparing with those of the control were 21.20%+/-1.62% vs 2.24%+/-0.26% and 21.23+/-1.78 vs 2.01+/-0.23 (P【0.05). CONCLUSION:The selective COX-2 inhibitor, Nimesulide can inhibit the proliferation of SMMC-7721 cells and increase apoptosis rate and apoptosis index of SMMC-7721 cells. The apoptosis rate and the apoptosis index are dose-dependent. Under electron microscope SMMC-7721 cells incubated with 300 micromol and 400 micromol Nimesulide show apoptotic characteristics. With the clarification of the mechanism of selective COX-2 inhibitors, These COX-2 selective inhibitors can become the choice of prevention and treatment of cancers.展开更多
AIM:To study the relationship between the cyclooxy-genase(COX)-2 gene and the proliferation and apopto-sis of esophageal squamous carcinoma EC109 cells.METHODS:The techniques of RNA interference(RNAi)and cell transfec...AIM:To study the relationship between the cyclooxy-genase(COX)-2 gene and the proliferation and apopto-sis of esophageal squamous carcinoma EC109 cells.METHODS:The techniques of RNA interference(RNAi)and cell transfection,as well as the levels of oncogenic-ity in nude mice,were used to study the role of COX-2 in the esophageal squamous carcinoma cell(ESCC)line EC109.Following RNAi and transfection,Western blot-ting analysis was used to determine the expression of the COX-2 protein.The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide(MTT)reduction assay was used to evaluate cell growth,and flow cytometry was used to detect cell apoptosis.RESULTS:Western blotting analysis demonstrated that COX-2 expression was significantly reduced in EC109 cells treated with COX-2-specif ic short interfering RNA(siRNA)but was increased in EC109 cells transfected with COX-2.Furthermore,COX-2 siRNA treatment in-hibited cell proliferation(P < 0.01)and induced apop-tosis in EC109 cells,as determined by an MTT assay and by flow cytometry,respectively.In contrast,trans-fected COX-2 led to increased cell proliferation(P < 0.05)and decreased apoptosis in EC109 cells.In addition,combination treatment of cells with COX-2 siRNA and aspirin had a synergistic effect(P < 0.01).For experi-ments measuring tumorigenicity,xenograft tumors of a greater volume and weight were found in the COX-2 group compared with other groups(P < 0.05).A large dose of aspirin inhibited tumor growth in nude mice ef-fectively(P < 0.05),and the rate of tumor suppression was 51.8% in the high-dose aspirin group.CONCLUSION:COX-2 plays a very critical role in ESCC carcinogenesis,and COX-2 siRNA combined with aspirin has the potential to be an anticancer therapy for the treatment of ESCC.展开更多
ABM: Recent studies suggested that cyclooxygenase-2 (COX-2) enhances tumor angiogenesis via upregulation of vascular endothelial growth factor (VEGF). Although COX-2 expression has been demonstrated in hepatocellular ...ABM: Recent studies suggested that cyclooxygenase-2 (COX-2) enhances tumor angiogenesis via upregulation of vascular endothelial growth factor (VEGF). Although COX-2 expression has been demonstrated in hepatocellular carcinoma (HCC), the significance of COX-2 in progression of HCC remains unclear. This study evaluated the clinico-pathological correlation of COX-2 level and its relationship with VEGF level in HCC. METHODS: Fresh tumor tissues were obtained from 100 patients who underwent resection of HCC. COX-2 protein expression was examined by immunohistochemistry, and quantitatively by an enzyme immunometric assay (EIA) of tumor cytosolic COX-2 levels. Tumor cytosolic VEGF levels were measured by an ELISA. RESULTS: Immunostaining showed expression of COX-2 in tumor cells. Tumor cytosolic COX-2 levels correlated with VEGF levels (r = 0.469,P<0.001). Correlation with clinicopathological features showed significantly higher tumor cytosolic COX-2 levels in the presence of multiple tumors (P = 0.027), venous invasion (P = 0.030), microsatellite lesions (P=0.037) and advanced tumor stage (P = 0.008). Higher tumor cytosolic COX-2 levels were associated with worse patient survival. CONCLUSION: This study shows that elevated tumor COX-2 levels correlate with elevated VEGF levels and invasiveness in HCC, suggesting that COX-2 plays a significant role in the progression of HCC.展开更多
AIM: To investigate the relation between the expression of cyclooxygenase-2 (COX-2) and liver cancer, to construct the recombinant adenovirus encoding human COX-2antisense RNA, and to explore its effects on liver canc...AIM: To investigate the relation between the expression of cyclooxygenase-2 (COX-2) and liver cancer, to construct the recombinant adenovirus encoding human COX-2antisense RNA, and to explore its effects on liver cancer cell proliferation.METHODS: We studied the expression of COX-2 in 34cases of hepatocellular carcinoma (HCC) and SMMC7402and SMMC7721 by immunohistochemical technique.Recombinant adenovirus Ad-AShcox-2 was constructed and transfected into human HCC cell lines SMMC7402and SMMC7721, and its effects on COX-2 expression, cell apoptosis and cell cycle were analyzed by flow cytometry.Cell proliferation was determined by colony-forming efficiency.RESULTS: We observed COX-2 expression in 82.4% of HCC and SMMC7402 cells, but no COX-2 expression in SMMC7721 cells. In addition, recombinant adenovirus encoding antisense COX-2 fragment Ad-AShcox-2 was obtained with the titer of 1.06× 1012 PFU/mL. Ad-AShcox-2 could reduce the expression of COX-2 and enhance the percentage of cells in G1/G0 phase in SMMC7402 cell line.The difference of apoptotic index between the Ad-AShcox2 group and control group was statistically significant(tcontrol group= 32.62 and tAd-LacZ= 10.93, P＜0.001) in SMMC7402 but not in SMMC7721. Similarly, colony-forming rates of SMMC7402 and SMMC7721 cell lines, after the transfer of Ad-AShcox-2, were (2.7±0.94)% and(33.6±4.24)%, respectively.CONCLUSION: Reduction in the expression of COX-2 can inhibit COX-2 expressing HCC cells.展开更多
Hepatitis B virus X protein （HBx） plays a crucial role in the development of hepatocellular carcinoma. Here, we sought to identify the mechanisms by which HBx mediates liver cell proliferation. We found that HBx upr...Hepatitis B virus X protein （HBx） plays a crucial role in the development of hepatocellular carcinoma. Here, we sought to identify the mechanisms by which HBx mediates liver cell proliferation. We found that HBx upregulated the levels of cyclooxygenase-2 （COX-2）, 5-1ipoxygenase （5-LOX） and phosphorylated extracellular signal-regulated protein kinases 1/2 （p-ERK1/2） in liver cells. HBx-induced p-ERK1/2 was abolished by inhibition of Gi/o proteins, COX or LOX. In addition, HBx increased the amounts of prostaglandin E2 （PGE2） and leukotriene B4 （LTB4） released from cell lines derived from hepatocytes. Moreover, these released arachidonic acid metabolites were able to activate ERK1/2. Interestingly, activated ERK1/2 could upregulate the expression of COX-2 and 5-LOX in a positive feedback manner. In conclusion, HBx enhances and maintains liver cell proliferation via a positive feedback loop involving COX-2, 5-LOX, released arachidonic acid metabolites, Gi/o proteins and p-ERK1/2.展开更多
Hepatic stellate cells(HSCs) are a kind of adipocytes. In HSCs lipids mainly exist in the form of lipid droplets. They are abundantly found in the cytoplasm and their main constituents are triglycerides. Lipid metabol...Hepatic stellate cells(HSCs) are a kind of adipocytes. In HSCs lipids mainly exist in the form of lipid droplets. They are abundantly found in the cytoplasm and their main constituents are triglycerides. Lipid metabolism in HSCs is closely related to its biological activity, however the mechanism of lipid droplets disappearance after HSC activation is not clearly established yet. Recent research shows that, cyclooxygenase-2 plays an important regulatory role in the lipid metabolism of HSCs. This paper seeks to review the subject based on studies that have been conducted so far to understand the role of cyclooxygenase-2 in the metabolism of lipids in HSCs.展开更多
Objective To investigate the expression of cyclooxygenase-2 （ Cox-2 ） and microvessel density （MVD） in benign and malignant pheochromocytomas, and the relationship between Cox-2 and MVD. Methods Specimens and clin...Objective To investigate the expression of cyclooxygenase-2 （ Cox-2 ） and microvessel density （MVD） in benign and malignant pheochromocytomas, and the relationship between Cox-2 and MVD. Methods Specimens and clinical data from 38 patients （ 21 benign and 17 malignant pheochromocytomas ） were studied. Slides of normal adrenal glands in nephrectomy specimens from another 20 patients with benign renal tumors were used as control. Irnmunohistochemical technology was performed to detect the Cox-2 and MVD in all specimens. Results Expression of Cox-2 was observed in 5 of the 21 benign pheochromocytomas （23. 8% ） , and in 14 of the 17 malignant （82.4%）. No expression of Cox-2 was observed in control slides. There were significant differences of Cox-2 expression between benign and malignant pheochromocytomas, as well as between malignant pheochromocytomas and control （ P 〈0. 05）. Expressions of MVD were 36. 41 ±13. 00, 21.43 ±8. 05, and 13. 36 ±4.34 in malignant, benign pheochromocytomas, and in control, respectively. Conclusion Cox-2 may contribute to the invasive characteristics of malignant pheochromocytomas and be used as a marker to distinguish malignant from benign pheochromocytomas. Expression of MVD in malignant pheochromocytomas was directly correlated with Cox-2.展开更多
Blueberries are a rich source of anthocyanins, which are associated with health benefits contributing to a reduced risk for many diseases. The present study identified the functional Gardenblue blueberry （Vaccinium a...Blueberries are a rich source of anthocyanins, which are associated with health benefits contributing to a reduced risk for many diseases. The present study identified the functional Gardenblue blueberry （Vaccinium ashei Reade） anthocyanin extracts （GBBAEs） and evaluated their capacity and underlying mechanisms in protecting murine RAW 264.7 cells from lipopolysaccharide （LPS）-stimulated inflammation in vitro. Enzyme-linked immunosorbent assay （ELISA） kit results showed that GBBAEs significantly inhibited the production of nitric oxide （NO）, prostaglandin E2 （PGE2）, interleukin-6 （IL-6）, IL-113, and interferon-y （INF-y）. Real-time polymerase chain reaction （PCR） analysis in- dicated that the mRNA expression levels of IL-6, IL-113, tumor necrosis factor-α （TNF-α）, monocyte chemoattractant protein-1 （MCP-1）, and cyclooxygenase 2 （COX-2） were suppressed in LPS-stimulated RAW 264.7 cells. Additionally, Western blot analysis was used to evaluate the relative protein expression levels of COX-2 and nuclear factor-κB p65 （NF-κBp65）. All these results suggested the potential use of GBBAEs as a functional food for the treatment of in- flammatory diseases.展开更多
文摘AIM: Cyclooxygenase-2 (COX-2) has been suggested to be associated with carcinogenesis. We sought to investigate the effect of the selective COX-2 inhibitor, Nimesulide on proliferation and apoptosis of SMMC-7721 human hepatoma cells.METHODS: This study was carried out on the culture of hepatic carcinoma SMMC-7721 cell line. Various concentrations of Nimesulide (0, 200 micromol/L, 300 micromol/L, 400 micromol/L) were added and incubated. Cell proliferation was detected with MTT colorimetric assay, cell apoptosis by electron microscopy, flow cytometry and TUNEL.RESULTS: Nimesulide could significantly inhibit SMMC-7721 cells proliferation dose-dependent and in a dependent manner compared with that of the control group. The duration lowest inhibition rate produced by Nimesulide in SMMC-7721 cells was 19.06%, the highest inhibition rate was 58.49%. After incubation with Nimesulide for 72 h, the most highest apoptosis rate and apoptosis index of SMMC-7721 cells comparing with those of the control were 21.20%+/-1.62% vs 2.24%+/-0.26% and 21.23+/-1.78 vs 2.01+/-0.23 (P【0.05). CONCLUSION:The selective COX-2 inhibitor, Nimesulide can inhibit the proliferation of SMMC-7721 cells and increase apoptosis rate and apoptosis index of SMMC-7721 cells. The apoptosis rate and the apoptosis index are dose-dependent. Under electron microscope SMMC-7721 cells incubated with 300 micromol and 400 micromol Nimesulide show apoptotic characteristics. With the clarification of the mechanism of selective COX-2 inhibitors, These COX-2 selective inhibitors can become the choice of prevention and treatment of cancers.
基金Supported by The National Natural Science Foundation of China,No.81071974The National 863 High Technology Research and Development Plan of China,No.2007AA02Z4Z4
文摘AIM:To study the relationship between the cyclooxy-genase(COX)-2 gene and the proliferation and apopto-sis of esophageal squamous carcinoma EC109 cells.METHODS:The techniques of RNA interference(RNAi)and cell transfection,as well as the levels of oncogenic-ity in nude mice,were used to study the role of COX-2 in the esophageal squamous carcinoma cell(ESCC)line EC109.Following RNAi and transfection,Western blot-ting analysis was used to determine the expression of the COX-2 protein.The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide(MTT)reduction assay was used to evaluate cell growth,and flow cytometry was used to detect cell apoptosis.RESULTS:Western blotting analysis demonstrated that COX-2 expression was significantly reduced in EC109 cells treated with COX-2-specif ic short interfering RNA(siRNA)but was increased in EC109 cells transfected with COX-2.Furthermore,COX-2 siRNA treatment in-hibited cell proliferation(P < 0.01)and induced apop-tosis in EC109 cells,as determined by an MTT assay and by flow cytometry,respectively.In contrast,trans-fected COX-2 led to increased cell proliferation(P < 0.05)and decreased apoptosis in EC109 cells.In addition,combination treatment of cells with COX-2 siRNA and aspirin had a synergistic effect(P < 0.01).For experi-ments measuring tumorigenicity,xenograft tumors of a greater volume and weight were found in the COX-2 group compared with other groups(P < 0.05).A large dose of aspirin inhibited tumor growth in nude mice ef-fectively(P < 0.05),and the rate of tumor suppression was 51.8% in the high-dose aspirin group.CONCLUSION:COX-2 plays a very critical role in ESCC carcinogenesis,and COX-2 siRNA combined with aspirin has the potential to be an anticancer therapy for the treatment of ESCC.
基金Supported by the Sun C.Y. Research Foundation for Hepatobiliary and Pancreatic Surgery of the University of Hong Kong
文摘ABM: Recent studies suggested that cyclooxygenase-2 (COX-2) enhances tumor angiogenesis via upregulation of vascular endothelial growth factor (VEGF). Although COX-2 expression has been demonstrated in hepatocellular carcinoma (HCC), the significance of COX-2 in progression of HCC remains unclear. This study evaluated the clinico-pathological correlation of COX-2 level and its relationship with VEGF level in HCC. METHODS: Fresh tumor tissues were obtained from 100 patients who underwent resection of HCC. COX-2 protein expression was examined by immunohistochemistry, and quantitatively by an enzyme immunometric assay (EIA) of tumor cytosolic COX-2 levels. Tumor cytosolic VEGF levels were measured by an ELISA. RESULTS: Immunostaining showed expression of COX-2 in tumor cells. Tumor cytosolic COX-2 levels correlated with VEGF levels (r = 0.469,P<0.001). Correlation with clinicopathological features showed significantly higher tumor cytosolic COX-2 levels in the presence of multiple tumors (P = 0.027), venous invasion (P = 0.030), microsatellite lesions (P=0.037) and advanced tumor stage (P = 0.008). Higher tumor cytosolic COX-2 levels were associated with worse patient survival. CONCLUSION: This study shows that elevated tumor COX-2 levels correlate with elevated VEGF levels and invasiveness in HCC, suggesting that COX-2 plays a significant role in the progression of HCC.
基金Supported by the Bureau of Science and Technology of Shiyan City
文摘AIM: To investigate the relation between the expression of cyclooxygenase-2 (COX-2) and liver cancer, to construct the recombinant adenovirus encoding human COX-2antisense RNA, and to explore its effects on liver cancer cell proliferation.METHODS: We studied the expression of COX-2 in 34cases of hepatocellular carcinoma (HCC) and SMMC7402and SMMC7721 by immunohistochemical technique.Recombinant adenovirus Ad-AShcox-2 was constructed and transfected into human HCC cell lines SMMC7402and SMMC7721, and its effects on COX-2 expression, cell apoptosis and cell cycle were analyzed by flow cytometry.Cell proliferation was determined by colony-forming efficiency.RESULTS: We observed COX-2 expression in 82.4% of HCC and SMMC7402 cells, but no COX-2 expression in SMMC7721 cells. In addition, recombinant adenovirus encoding antisense COX-2 fragment Ad-AShcox-2 was obtained with the titer of 1.06× 1012 PFU/mL. Ad-AShcox-2 could reduce the expression of COX-2 and enhance the percentage of cells in G1/G0 phase in SMMC7402 cell line.The difference of apoptotic index between the Ad-AShcox2 group and control group was statistically significant(tcontrol group= 32.62 and tAd-LacZ= 10.93, P＜0.001) in SMMC7402 but not in SMMC7721. Similarly, colony-forming rates of SMMC7402 and SMMC7721 cell lines, after the transfer of Ad-AShcox-2, were (2.7±0.94)% and(33.6±4.24)%, respectively.CONCLUSION: Reduction in the expression of COX-2 can inhibit COX-2 expressing HCC cells.
文摘Hepatitis B virus X protein （HBx） plays a crucial role in the development of hepatocellular carcinoma. Here, we sought to identify the mechanisms by which HBx mediates liver cell proliferation. We found that HBx upregulated the levels of cyclooxygenase-2 （COX-2）, 5-1ipoxygenase （5-LOX） and phosphorylated extracellular signal-regulated protein kinases 1/2 （p-ERK1/2） in liver cells. HBx-induced p-ERK1/2 was abolished by inhibition of Gi/o proteins, COX or LOX. In addition, HBx increased the amounts of prostaglandin E2 （PGE2） and leukotriene B4 （LTB4） released from cell lines derived from hepatocytes. Moreover, these released arachidonic acid metabolites were able to activate ERK1/2. Interestingly, activated ERK1/2 could upregulate the expression of COX-2 and 5-LOX in a positive feedback manner. In conclusion, HBx enhances and maintains liver cell proliferation via a positive feedback loop involving COX-2, 5-LOX, released arachidonic acid metabolites, Gi/o proteins and p-ERK1/2.
基金Supported by the National Natural Science Foundation of China(81373465)
文摘Hepatic stellate cells(HSCs) are a kind of adipocytes. In HSCs lipids mainly exist in the form of lipid droplets. They are abundantly found in the cytoplasm and their main constituents are triglycerides. Lipid metabolism in HSCs is closely related to its biological activity, however the mechanism of lipid droplets disappearance after HSC activation is not clearly established yet. Recent research shows that, cyclooxygenase-2 plays an important regulatory role in the lipid metabolism of HSCs. This paper seeks to review the subject based on studies that have been conducted so far to understand the role of cyclooxygenase-2 in the metabolism of lipids in HSCs.
文摘Objective To investigate the expression of cyclooxygenase-2 （ Cox-2 ） and microvessel density （MVD） in benign and malignant pheochromocytomas, and the relationship between Cox-2 and MVD. Methods Specimens and clinical data from 38 patients （ 21 benign and 17 malignant pheochromocytomas ） were studied. Slides of normal adrenal glands in nephrectomy specimens from another 20 patients with benign renal tumors were used as control. Irnmunohistochemical technology was performed to detect the Cox-2 and MVD in all specimens. Results Expression of Cox-2 was observed in 5 of the 21 benign pheochromocytomas （23. 8% ） , and in 14 of the 17 malignant （82.4%）. No expression of Cox-2 was observed in control slides. There were significant differences of Cox-2 expression between benign and malignant pheochromocytomas, as well as between malignant pheochromocytomas and control （ P 〈0. 05）. Expressions of MVD were 36. 41 ±13. 00, 21.43 ±8. 05, and 13. 36 ±4.34 in malignant, benign pheochromocytomas, and in control, respectively. Conclusion Cox-2 may contribute to the invasive characteristics of malignant pheochromocytomas and be used as a marker to distinguish malignant from benign pheochromocytomas. Expression of MVD in malignant pheochromocytomas was directly correlated with Cox-2.
基金Project supported by the Fundamental Research Funds for the Central Universities of China(No.2013PY095)the Clinical Research Project of Health and Family Planning Commission of Wuhan Municipality(No.WX13A05)+1 种基金the Research Project of Wuhan City Central Hospital(No.YQ15A04)the Grant from Key Laboratory of Biological Targeted Therapy of Hubei Province of China(No.02.03.2014-10)
文摘Blueberries are a rich source of anthocyanins, which are associated with health benefits contributing to a reduced risk for many diseases. The present study identified the functional Gardenblue blueberry （Vaccinium ashei Reade） anthocyanin extracts （GBBAEs） and evaluated their capacity and underlying mechanisms in protecting murine RAW 264.7 cells from lipopolysaccharide （LPS）-stimulated inflammation in vitro. Enzyme-linked immunosorbent assay （ELISA） kit results showed that GBBAEs significantly inhibited the production of nitric oxide （NO）, prostaglandin E2 （PGE2）, interleukin-6 （IL-6）, IL-113, and interferon-y （INF-y）. Real-time polymerase chain reaction （PCR） analysis in- dicated that the mRNA expression levels of IL-6, IL-113, tumor necrosis factor-α （TNF-α）, monocyte chemoattractant protein-1 （MCP-1）, and cyclooxygenase 2 （COX-2） were suppressed in LPS-stimulated RAW 264.7 cells. Additionally, Western blot analysis was used to evaluate the relative protein expression levels of COX-2 and nuclear factor-κB p65 （NF-κBp65）. All these results suggested the potential use of GBBAEs as a functional food for the treatment of in- flammatory diseases.
