AIM: To investigate the effects of the WWOX gene on the human hepatic carcinoma cell line SMMC-7721. METHODS: Full-length WWOX cDNA was amplified from normal human liver tissues. Full-length cDNA was subcloned into pE...AIM: To investigate the effects of the WWOX gene on the human hepatic carcinoma cell line SMMC-7721. METHODS: Full-length WWOX cDNA was amplified from normal human liver tissues. Full-length cDNA was subcloned into pEGFP-N1, a eukaryotic expression vector. After introduction of the WWOX gene into cancer cells using liposomes, the WWOX protein level in the cells was detected through Western blotting. Cell growth rates were assessed by methyl thiazolyl tetrazolium (MTT) and colony formation assays. Cell cycle progression and cell apoptosis were measured by flow cytometry. The phosphorylated protein kinase B (AKT) and activated fragments of caspase-9 and caspase-3 were examined by Western blotting analysis. RESULTS: WWOX significantly inhibited cell proliferation, as evaluated by the MTT and colony formation assays. Cells transfected with WWOX showed significantly higher apoptosis ratios when compared with cells transfected with a mock plasmid, and overexpression of WWOX delayed cell cycle progression from G1 to S phase, as measured by flow cytometry. An increase in apoptosis was also indicated by a remarkable activation of caspase-9 and caspase-3 and a dephosphorylation of AKT (Thr308 and Ser473) measured with Western blotting analysis. CONCLUSION: Overexpression of WWOX induces apoptosis and inhibits proliferation of the human hepatic carcinoma cell line SMMC-7721.展开更多
OBJECTIVE:To study the possible roles of Jinlong capsule(JLC) on the proliferation and apoptosis of human pancreatic cancer cells BxPC-3.METHODS:The human pancreatic cancer cells BxPC-3 were treated with JLC at the co...OBJECTIVE:To study the possible roles of Jinlong capsule(JLC) on the proliferation and apoptosis of human pancreatic cancer cells BxPC-3.METHODS:The human pancreatic cancer cells BxPC-3 were treated with JLC at the concentration of 0.05-1.00 mg/mL for 24-120 h.The inhibition rate of JLC on human pancreatic cancer cells BxPC-3 was detected by 3-(4,5-dimethiylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) assay.Flow cytometry was employed to measure cell apoptosis using Annexin V-FITC/Propidium iodide(AV-FITC/PI) method.Cell cycles were determined by PI staining.The expression of S100 Calcium binding protein A4(S100A4) in cell matrix was measured by enzyme-linked immunosorbent assay(ELISA).The expression levels of apoptosis-related protein such as BCL2/adenovirus E1B 19 kDa interacting protein 3(BNIP3),B-cell lymphoma/leukemia-2(Bcl-2) and Cys-teinylaspartate specific proteinase 3(Caspase-3) were detected byWestern blotting.RESULTS:JLC significantly inhibited the proliferation of human pancreatic cancer cells BxPC-3 in a dose-dependent and time-dependent manner.JLC promoted cell apoptosis and maintained cell cycle in S and G 2 /M phase rather than G 1 /G 0 phase.The expression of S100A4 in the cell matrix was reduced.The expression of cell apoptotic protein BNIP3 was increased while Bcl-2 was decreased.CONCLUSION:JLC can inhibit the proliferation of human pancreatic cancer cells BxPC-3 by stimulating cell apoptosis,arresting the cell cycle at S and G 2 /M phase which blocks the circulation of normal cell cycle and reducing the expression of S100A4 protein.Higher pro-apoptosis protein BNIP3 and lower anti-apoptosis protein Bcl-2 levels were found,which may be related to the apoptotic effects of JLC.展开更多
目的研究桂皮醛对激素诱导的体外破骨细胞增殖分化及骨吸收功能增强的保护作用及分子机制。方法RANKL联合M-CSF体外诱导RAW264.7细胞分化为破骨细胞,分为对照组、地塞米松处理组和不同浓度(11.6、23.2、46.4μg·L-1)桂皮醛(cinnami...目的研究桂皮醛对激素诱导的体外破骨细胞增殖分化及骨吸收功能增强的保护作用及分子机制。方法RANKL联合M-CSF体外诱导RAW264.7细胞分化为破骨细胞,分为对照组、地塞米松处理组和不同浓度(11.6、23.2、46.4μg·L-1)桂皮醛(cinnamic alde byde,CA)干预组;TRAP染色试剂盒鉴定成熟破骨细胞;MTT法观察地塞米松和桂皮醛在不同时间点对破骨细胞增殖的抑制作用;ELISA法检测细胞培养液上清中TRACP5b的表达水平;RT-PCR分析破骨细胞相关转录因子(RANK、NFATc1)mRNA的表达状态。结果地塞米松处理后,破骨细胞的增殖分化及骨吸收功能明显增强(P<0.05);而经不同浓度桂皮醛干预后,与地塞米松组相比,细胞增殖活性、上清液中TRACP-5b的含量以及RANK、NFATc1mRNA的表达水平随药物浓度的增加呈剂量依赖性降低(P<0.05)。结论桂皮醛可有效抑制激素诱导的破骨细胞的增殖及骨吸收功能,其作用机制可能是通过下调RANK以及下游NFATc1基因的表达。展开更多
文摘AIM: To investigate the effects of the WWOX gene on the human hepatic carcinoma cell line SMMC-7721. METHODS: Full-length WWOX cDNA was amplified from normal human liver tissues. Full-length cDNA was subcloned into pEGFP-N1, a eukaryotic expression vector. After introduction of the WWOX gene into cancer cells using liposomes, the WWOX protein level in the cells was detected through Western blotting. Cell growth rates were assessed by methyl thiazolyl tetrazolium (MTT) and colony formation assays. Cell cycle progression and cell apoptosis were measured by flow cytometry. The phosphorylated protein kinase B (AKT) and activated fragments of caspase-9 and caspase-3 were examined by Western blotting analysis. RESULTS: WWOX significantly inhibited cell proliferation, as evaluated by the MTT and colony formation assays. Cells transfected with WWOX showed significantly higher apoptosis ratios when compared with cells transfected with a mock plasmid, and overexpression of WWOX delayed cell cycle progression from G1 to S phase, as measured by flow cytometry. An increase in apoptosis was also indicated by a remarkable activation of caspase-9 and caspase-3 and a dephosphorylation of AKT (Thr308 and Ser473) measured with Western blotting analysis. CONCLUSION: Overexpression of WWOX induces apoptosis and inhibits proliferation of the human hepatic carcinoma cell line SMMC-7721.
文摘OBJECTIVE:To study the possible roles of Jinlong capsule(JLC) on the proliferation and apoptosis of human pancreatic cancer cells BxPC-3.METHODS:The human pancreatic cancer cells BxPC-3 were treated with JLC at the concentration of 0.05-1.00 mg/mL for 24-120 h.The inhibition rate of JLC on human pancreatic cancer cells BxPC-3 was detected by 3-(4,5-dimethiylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) assay.Flow cytometry was employed to measure cell apoptosis using Annexin V-FITC/Propidium iodide(AV-FITC/PI) method.Cell cycles were determined by PI staining.The expression of S100 Calcium binding protein A4(S100A4) in cell matrix was measured by enzyme-linked immunosorbent assay(ELISA).The expression levels of apoptosis-related protein such as BCL2/adenovirus E1B 19 kDa interacting protein 3(BNIP3),B-cell lymphoma/leukemia-2(Bcl-2) and Cys-teinylaspartate specific proteinase 3(Caspase-3) were detected byWestern blotting.RESULTS:JLC significantly inhibited the proliferation of human pancreatic cancer cells BxPC-3 in a dose-dependent and time-dependent manner.JLC promoted cell apoptosis and maintained cell cycle in S and G 2 /M phase rather than G 1 /G 0 phase.The expression of S100A4 in the cell matrix was reduced.The expression of cell apoptotic protein BNIP3 was increased while Bcl-2 was decreased.CONCLUSION:JLC can inhibit the proliferation of human pancreatic cancer cells BxPC-3 by stimulating cell apoptosis,arresting the cell cycle at S and G 2 /M phase which blocks the circulation of normal cell cycle and reducing the expression of S100A4 protein.Higher pro-apoptosis protein BNIP3 and lower anti-apoptosis protein Bcl-2 levels were found,which may be related to the apoptotic effects of JLC.
文摘目的研究桂皮醛对激素诱导的体外破骨细胞增殖分化及骨吸收功能增强的保护作用及分子机制。方法RANKL联合M-CSF体外诱导RAW264.7细胞分化为破骨细胞,分为对照组、地塞米松处理组和不同浓度(11.6、23.2、46.4μg·L-1)桂皮醛(cinnamic alde byde,CA)干预组;TRAP染色试剂盒鉴定成熟破骨细胞;MTT法观察地塞米松和桂皮醛在不同时间点对破骨细胞增殖的抑制作用;ELISA法检测细胞培养液上清中TRACP5b的表达水平;RT-PCR分析破骨细胞相关转录因子(RANK、NFATc1)mRNA的表达状态。结果地塞米松处理后,破骨细胞的增殖分化及骨吸收功能明显增强(P<0.05);而经不同浓度桂皮醛干预后,与地塞米松组相比,细胞增殖活性、上清液中TRACP-5b的含量以及RANK、NFATc1mRNA的表达水平随药物浓度的增加呈剂量依赖性降低(P<0.05)。结论桂皮醛可有效抑制激素诱导的破骨细胞的增殖及骨吸收功能,其作用机制可能是通过下调RANK以及下游NFATc1基因的表达。
