采用DEAE-FF离子交换和Sephdax G-15葡聚糖凝胶层析方法,从玉米肽中分离、纯化,获得具有抑制HepG2细胞增殖活性的短肽,采用高效液相色谱串联质谱电喷雾法(liquid chromatography-electrospray ionization tandem mass spectrometry,LC-E...采用DEAE-FF离子交换和Sephdax G-15葡聚糖凝胶层析方法,从玉米肽中分离、纯化,获得具有抑制HepG2细胞增殖活性的短肽,采用高效液相色谱串联质谱电喷雾法(liquid chromatography-electrospray ionization tandem mass spectrometry,LC-ESI-MS/MS)对其活性片段进行结构鉴定。结果表明:其短肽的一级结构为LPPYLP[命名为玉米六肽(corn peptides-6,CPs-6)],通过四唑盐[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT]比色法试验发现,CPs-6具有抑制HepG2细胞增殖的效果,且对L-02细胞的毒性低。展开更多
AIM：To investigate the cytotoxic effects of spray-dried extracts of Phyllanthus niruri in combination with cis- platin on two cancer cell lines. METHODS： Colorectal carcinoma （HT29） and human hepatocellular carcin...AIM：To investigate the cytotoxic effects of spray-dried extracts of Phyllanthus niruri in combination with cis- platin on two cancer cell lines. METHODS： Colorectal carcinoma （HT29） and human hepatocellular carcinoma （HepG2） cells were treated with spray-dried extracts of Phyllanthus niruri （SDEPN） either alone or in combination with cisplatin at differ- ent concentrations （0.5 mg/mL and 1 mg/mL） for 4 h and 24 h. To verify and quantify cancer cells treated with these products as well as identify the cell cycle stage and cell viability, we stained the cells with prop- idium iodide and assessed them by flow cytometry. The percentage of cells in different cell cycle phases was quantified and data were expressed as histo- grams. Significant differences between groups were determined using analysis of variance and Bonferroni＇s test, as indicated. A value of P 〈 0.05 was considered to be statistically significant. RESULTS： SDEPN had significantly different cyto- toxic effects on HT29 （2.81 4- 0.11 vs 3.51 4- 1.13, P 〉 0.05） and HepG2 （5.07± 0.3 vs 15.9 ± 1.04, P 〈 0.001） cells when compared to control cells for 4 h. SDEPN also had significantly different cytotoxic effects on HT29 （1.91 ± 0.57 vs 4.53± 1.22, P 〉 0.05） and HepG2 （14.56 ± 1.6 vs 35.67 ± 3.94, P 〈 0.001） cells when compared to control cells for 24 h. Both cell lines were killed by cisplatin in a dose-dependent manner compared to control cells （HepG2 cells for 4 h： 10.78 ± 1.58 vs 53.89 ± 1.53, P 〈 0.001; 24 h： 8.9 ± 1.43 vs 62.78 ± 1.87, P 〈 0.001 and HT29 cells for 4 h： 9.52 ±0.913 vs 49.86 ± 2.89, P 〈 0.001; 24 h： 11.78 ± 1.05 vs 53.34 ± 2.65, P 〈 0.001）. In HT29 cells, pretreat- ment with SDEPN and subsequent treatment with cis-platin resulted in a greater number of cells being killed （12.78 ± 1.01 vs 93.76 ± 1.6, P 〈 0.001）. HepG2 cells showed significant cell killing with treatment with SDEPN when combined with cisplatin （12.87 ± 2.78 vs 78.8 ± 3.02, P 〈 0.001）. CONCLUSION： SDEPN is selectively toxic against two cancer cell lines. Moreover, SDEPN in combination with cisplatin induces a synergistic increase in the cell death of both HT29 and HepG2 cells.展开更多
文摘采用DEAE-FF离子交换和Sephdax G-15葡聚糖凝胶层析方法,从玉米肽中分离、纯化,获得具有抑制HepG2细胞增殖活性的短肽,采用高效液相色谱串联质谱电喷雾法(liquid chromatography-electrospray ionization tandem mass spectrometry,LC-ESI-MS/MS)对其活性片段进行结构鉴定。结果表明:其短肽的一级结构为LPPYLP[命名为玉米六肽(corn peptides-6,CPs-6)],通过四唑盐[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT]比色法试验发现,CPs-6具有抑制HepG2细胞增殖的效果,且对L-02细胞的毒性低。
基金Supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)CNPq (470179/2009-0) for financial support and Postgraduate Program in Pharmaceutical Sciences,Federal University of Rio Grande do Norte
文摘AIM：To investigate the cytotoxic effects of spray-dried extracts of Phyllanthus niruri in combination with cis- platin on two cancer cell lines. METHODS： Colorectal carcinoma （HT29） and human hepatocellular carcinoma （HepG2） cells were treated with spray-dried extracts of Phyllanthus niruri （SDEPN） either alone or in combination with cisplatin at differ- ent concentrations （0.5 mg/mL and 1 mg/mL） for 4 h and 24 h. To verify and quantify cancer cells treated with these products as well as identify the cell cycle stage and cell viability, we stained the cells with prop- idium iodide and assessed them by flow cytometry. The percentage of cells in different cell cycle phases was quantified and data were expressed as histo- grams. Significant differences between groups were determined using analysis of variance and Bonferroni＇s test, as indicated. A value of P 〈 0.05 was considered to be statistically significant. RESULTS： SDEPN had significantly different cyto- toxic effects on HT29 （2.81 4- 0.11 vs 3.51 4- 1.13, P 〉 0.05） and HepG2 （5.07± 0.3 vs 15.9 ± 1.04, P 〈 0.001） cells when compared to control cells for 4 h. SDEPN also had significantly different cytotoxic effects on HT29 （1.91 ± 0.57 vs 4.53± 1.22, P 〉 0.05） and HepG2 （14.56 ± 1.6 vs 35.67 ± 3.94, P 〈 0.001） cells when compared to control cells for 24 h. Both cell lines were killed by cisplatin in a dose-dependent manner compared to control cells （HepG2 cells for 4 h： 10.78 ± 1.58 vs 53.89 ± 1.53, P 〈 0.001; 24 h： 8.9 ± 1.43 vs 62.78 ± 1.87, P 〈 0.001 and HT29 cells for 4 h： 9.52 ±0.913 vs 49.86 ± 2.89, P 〈 0.001; 24 h： 11.78 ± 1.05 vs 53.34 ± 2.65, P 〈 0.001）. In HT29 cells, pretreat- ment with SDEPN and subsequent treatment with cis-platin resulted in a greater number of cells being killed （12.78 ± 1.01 vs 93.76 ± 1.6, P 〈 0.001）. HepG2 cells showed significant cell killing with treatment with SDEPN when combined with cisplatin （12.87 ± 2.78 vs 78.8 ± 3.02, P 〈 0.001）. CONCLUSION： SDEPN is selectively toxic against two cancer cell lines. Moreover, SDEPN in combination with cisplatin induces a synergistic increase in the cell death of both HT29 and HepG2 cells.
