Objective： We have previously found that mbr is a regulatory element of the bcl2 gene. The objective of this study is to isolate and identify the proteins binding to the 37 mbr in the 3 ＇ -end of the mbr. Methods： ...Objective： We have previously found that mbr is a regulatory element of the bcl2 gene. The objective of this study is to isolate and identify the proteins binding to the 37 mbr in the 3 ＇ -end of the mbr. Methods： Streptavidin magnetic particles were ligated to concatameric oligonucleotides of 37 mbr and incubated with the nuclear extracts of Jurkat cells. The DNA-binding proteins were eluted and then resolved by SDS-PAGE. After silver staining, the protein bands were excised and subjected to MALDI-TOF MS. Results： Several protein bands were detected after the isolation with magnetic particles, and Splicing factor, proline- and glutamine-rich（SFPQ）, Poly（ADP-ribose） polymerase I（PARP）, and promyelocytic leukemia protein（PML） were identified by MALDI-TOF MS. Conclusion： Several proteins were isolated and identified from the 37 mbr-protein complex. Results of this study establish a foundation for further study of the mechanisms by which mbr executes its regulatory function.展开更多
Objectives:Although many studies have suggested the anticancer properties of Galium verum,there is still no accurate information regarding its side effects on normal cells.Accordingly,this study aimed to investigate t...Objectives:Although many studies have suggested the anticancer properties of Galium verum,there is still no accurate information regarding its side effects on normal cells.Accordingly,this study aimed to investigate the dual effects of the whole Galium verummethanolic extract on the normal human fibroblast cell line(AGO)cell line at different concentrations.Methods:The cell line was randomly divided into a control group and groups exposed to concentrations of 12.5 to 400μg/mL.Extraction was performed by the maceration method.In addition,the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide(MTT)method was applied to measure cytotoxicity and flow cytometry.Further,BCL2 associated X(BAX)andB-cell lymphoma 2(BCL2)genes were expressed by the real-time polymerase chain reaction to evaluate apoptosis and reactive oxygen species(ROS).Finally,data were compared between groups using a one-way analysis of variance.Results:A significant reduction was observed in the cell viability of 90%at a concentration of 400μg/mL compared to the control.In comparison,a significant increase was reported in cell viability at concentrations of 25-200μg/mL(P<0.0001).Furthermore,there was a significant 2.87-time increase in apoptosis compared to the control group(P<0.0001),but no significant differences were reported in cellular phases.ROS increased significantly by 5.7 times(P<0.05),and a significant 80-fold increase was found in the BAX/BCL2gene ratio(P<0.05).Conclusion:The whole methanolic extract could lower the viability of human fibroblasts at 400μg/mL and more by increasing apoptosis,thereby increasing BAX/BCL2gene expression and ROS production.However,the extract exerted an increased effect on cell viability in a concentration-dependent manner on AGO and increased cell growth at concentrations less than 400μg/mL,highlighting different effects of the whole extract on the AGO cell line.展开更多
Objective To study the expression of a novel inhibitor of apptosis and survivin in cervical carcinoma and its relationship to the expression of Bcl-2.Methods Using SP immunohistochemical technique, we examined the exp...Objective To study the expression of a novel inhibitor of apptosis and survivin in cervical carcinoma and its relationship to the expression of Bcl-2.Methods Using SP immunohistochemical technique, we examined the expression of survivin and Bcl 2 in 59 cervical invasive squamous cell carcinomas.Results Survivin was expressed in 41 of 59 cases(69.5%) of cervical carcinomas. In contrast, no expression of survivin in normal cervical tissues was observed. Overexpression of survivin was related to the tumor grade and clinical stage. Survivin positive cases were strongly associated with Bcl 2 expression(80% versus 35.7%; P <0.005).Conclusion Apoptosis inhibition by survivin abnormal expression, alone or in cooperation with Bcl 2, may participate in the onset and progression of cervical carcinoma. Survivin is a new diagnostic/therapeutic target in cervical cancer.展开更多
基金supported by grants from the National Natural Science Foundation of China(30500585)the Natural Science Foundation of Jiangsu Province(BK2008450)
文摘Objective： We have previously found that mbr is a regulatory element of the bcl2 gene. The objective of this study is to isolate and identify the proteins binding to the 37 mbr in the 3 ＇ -end of the mbr. Methods： Streptavidin magnetic particles were ligated to concatameric oligonucleotides of 37 mbr and incubated with the nuclear extracts of Jurkat cells. The DNA-binding proteins were eluted and then resolved by SDS-PAGE. After silver staining, the protein bands were excised and subjected to MALDI-TOF MS. Results： Several protein bands were detected after the isolation with magnetic particles, and Splicing factor, proline- and glutamine-rich（SFPQ）, Poly（ADP-ribose） polymerase I（PARP）, and promyelocytic leukemia protein（PML） were identified by MALDI-TOF MS. Conclusion： Several proteins were isolated and identified from the 37 mbr-protein complex. Results of this study establish a foundation for further study of the mechanisms by which mbr executes its regulatory function.
文摘Objectives:Although many studies have suggested the anticancer properties of Galium verum,there is still no accurate information regarding its side effects on normal cells.Accordingly,this study aimed to investigate the dual effects of the whole Galium verummethanolic extract on the normal human fibroblast cell line(AGO)cell line at different concentrations.Methods:The cell line was randomly divided into a control group and groups exposed to concentrations of 12.5 to 400μg/mL.Extraction was performed by the maceration method.In addition,the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide(MTT)method was applied to measure cytotoxicity and flow cytometry.Further,BCL2 associated X(BAX)andB-cell lymphoma 2(BCL2)genes were expressed by the real-time polymerase chain reaction to evaluate apoptosis and reactive oxygen species(ROS).Finally,data were compared between groups using a one-way analysis of variance.Results:A significant reduction was observed in the cell viability of 90%at a concentration of 400μg/mL compared to the control.In comparison,a significant increase was reported in cell viability at concentrations of 25-200μg/mL(P<0.0001).Furthermore,there was a significant 2.87-time increase in apoptosis compared to the control group(P<0.0001),but no significant differences were reported in cellular phases.ROS increased significantly by 5.7 times(P<0.05),and a significant 80-fold increase was found in the BAX/BCL2gene ratio(P<0.05).Conclusion:The whole methanolic extract could lower the viability of human fibroblasts at 400μg/mL and more by increasing apoptosis,thereby increasing BAX/BCL2gene expression and ROS production.However,the extract exerted an increased effect on cell viability in a concentration-dependent manner on AGO and increased cell growth at concentrations less than 400μg/mL,highlighting different effects of the whole extract on the AGO cell line.
文摘Objective To study the expression of a novel inhibitor of apptosis and survivin in cervical carcinoma and its relationship to the expression of Bcl-2.Methods Using SP immunohistochemical technique, we examined the expression of survivin and Bcl 2 in 59 cervical invasive squamous cell carcinomas.Results Survivin was expressed in 41 of 59 cases(69.5%) of cervical carcinomas. In contrast, no expression of survivin in normal cervical tissues was observed. Overexpression of survivin was related to the tumor grade and clinical stage. Survivin positive cases were strongly associated with Bcl 2 expression(80% versus 35.7%; P <0.005).Conclusion Apoptosis inhibition by survivin abnormal expression, alone or in cooperation with Bcl 2, may participate in the onset and progression of cervical carcinoma. Survivin is a new diagnostic/therapeutic target in cervical cancer.
