Objective： To investigate the inhibitory effect of interferon-α （IFN-α） and curcumin on proliferation of Raji cells （B-NHL） and its mechanism. Methods： The morphological, changes of Raji cells were observed in...Objective： To investigate the inhibitory effect of interferon-α （IFN-α） and curcumin on proliferation of Raji cells （B-NHL） and its mechanism. Methods： The morphological, changes of Raji cells were observed in culture medium with IFN-α （500, 1000, 2000, 3000 U/L） and various concentrations of curcumin （6.25, 12.5, 25 μmol/L） for different time in vitro. The inhibitory ratio was measured by MTT assay. Apoptosis was detected by flow cytometry （FCM）. The expression of caspase 6, caspase 8 and caspase 9 in Raji cells treated with IC5025 μmol/L curcumin with IFN-α was examined using Western blot. Results： IFN-α and curcumin could significantly inhibit the growth and induce apoptosis of RAji cells with synergistic effects. They could increase the expression of caspase 6, caspase 8 and caspase 9 in Raji cells in a dose- and time-dependent manner. Conclusion： The combined use of IFN-α and curcumin can inhibit the proliferation of B-NHL Raji cells apparently in vitro. Promotion of the expression of caspase 6, caspase 8, caspase 9 and induction of apoptosis might be one of the important mechanisms.展开更多
Inhibin a is one of the candidate genes that control the ovulation in poultry. To study the genetic effects of inhibin a on apoptosis and proliferation of goose granulosa cells cultured in vitro, two RNA interference ...Inhibin a is one of the candidate genes that control the ovulation in poultry. To study the genetic effects of inhibin a on apoptosis and proliferation of goose granulosa cells cultured in vitro, two RNA interference （RNAi） expression vectors, psiRNA-INHal and psiRNA-INHα2, were constructed to knock down inhibin α gene expression. After 48 h of transfection, the efficiency of these two RNAi expression vectors was examined by fluorescence microscopy. Meanwhile, inhibin protein expression levels, apoptosis indexes （AI） and proliferation indexes （PI） of granulosa cells were analyzed by flow cytometry. In addition, the supernatants were collected to assay the concentrations of estrogen （E2） and progesterone （P） by radioimmunoassay. The results showed that the expression level of inhibin a in the RNAi group were decreased 30%--40% than those in the control groups （P 〈0.05） and the apoptosis indexes and proliferation indexes in the RNAi groups were significantly higher than those in the control groups （P 〈0.05）. However, the E2 concentrations in the RNAi groups were lower than those in the control groups （P 〈0.05）. These results indicate that inhibin a has antagonistic effect on granulosa cell apoptosis.展开更多
Objective： We investigated the influence of CDK1 and CDK2 expression inhibited by cotransfection of CDK1 and CDK2 siRNA on cell cycle and apoptosis, explored the exact role of cell cycle master regulator in tumor cel...Objective： We investigated the influence of CDK1 and CDK2 expression inhibited by cotransfection of CDK1 and CDK2 siRNA on cell cycle and apoptosis, explored the exact role of cell cycle master regulator in tumor cell apoptosis process. Methods： The siRNA targeting the CDK1 and CDK2 genes were synthesized and simultaneously cotransfected into Hela cells by lipofectamine 2000.48 or 60 h after the cotransfection, CDK1 and CDK2 protein expressions were examined by Western blot. Cell cycle distribution was analyzed by flow cytometry. Cell apoptosis was detected by the Annexin V/PI method. The changes of the transfected cell morphological under a microscope after Wright-Giemsa Staining were studied. Results： CDK1 and CDK2 protein expression was decreased at 48 or 60 h after cotransfection. The accumulation of the G2/M and S phase population in cell cycle of the cotransfected cells at 48 or 60 h after transfection was enhanced obviously compared with control. The ratio of apoptotic cell of cotransfected cells at 48 or 60 h after transfection was increased significantly compared with control. More binucleate or multinucleate ceJls among cotransfected cells were observed under the microscope. Conclusion： The decreased expression of CDK1 and CDK2 by cotransfection of CDK1 and CDK2 siRNA not only leads to tumor cell cycle arrest in S phase and G2/M phase, but also induces tumor cell apoptosis.展开更多
Objective:The aim of this study was to investigate the molecular mechanism of anti-apoptotic action of survivin to the hepatoma-cellular carcinoma cell line HepG2.Methods:Design and synthesize siRNA gene sequence spec...Objective:The aim of this study was to investigate the molecular mechanism of anti-apoptotic action of survivin to the hepatoma-cellular carcinoma cell line HepG2.Methods:Design and synthesize siRNA gene sequence specifically targeting at HepG2 cell.HepG2 cells cultures were divided into five groups:blank control group,negative control group,low dose group,medium dose group and high dose group.HepG2 cells were treated respectively by pshRNA-survivin-387 of different concentrations.The apoptosis index(AI) was determined by flow cytometry(FCM).Cells were stained with rhodomine-123(Rh123) to detect changes of mitochondrial membrane potentials.The concentration of cytoplasmic cytochrome C(Cyt.C) was continuously determined by ELISA.Relative activities of caspase-9 and caspase-3 were assessed by colorimetric assay.Results:Compared with the control group,due to the function of short interference RNAs(SiRNAs) that suppresses the survivin gene expression,the apoptotic index of transfected groups were significantly higher than those of control groups(F = 13568.68,q = 110.47-327.16,P < 0.01),the apoptosis index of high concentration of transfected cells was higher than the low concentration transfected group(q = 39.63-168.22,P < 0.01).The apoptosis index of high concentrations transfected HepG2 cells was 25.54%,higher than that of blank control group,negative control group,low dose group and medium dose group(5.24%,6.61%,12.63% and 15.64%,respectively).HepG2 cells transfected with SiRNA exhibit gradually decreasing mitochondrial membrane potentials,which then lead to the releasing of Cyt C,following it were the activation of caspase-9 and caspase-3.Conclusion:Survivin performs the function of anti-apoptosis to the HepG2 cells via modulating the apoptosis of mitochondrial.HepG2 cells transfected with SiRNA survivin can significantly induce apoptosis.展开更多
基金This work was supported by a grant from National Natural Science Foundation of China (NO.30271672).
文摘Objective： To investigate the inhibitory effect of interferon-α （IFN-α） and curcumin on proliferation of Raji cells （B-NHL） and its mechanism. Methods： The morphological, changes of Raji cells were observed in culture medium with IFN-α （500, 1000, 2000, 3000 U/L） and various concentrations of curcumin （6.25, 12.5, 25 μmol/L） for different time in vitro. The inhibitory ratio was measured by MTT assay. Apoptosis was detected by flow cytometry （FCM）. The expression of caspase 6, caspase 8 and caspase 9 in Raji cells treated with IC5025 μmol/L curcumin with IFN-α was examined using Western blot. Results： IFN-α and curcumin could significantly inhibit the growth and induce apoptosis of RAji cells with synergistic effects. They could increase the expression of caspase 6, caspase 8 and caspase 9 in Raji cells in a dose- and time-dependent manner. Conclusion： The combined use of IFN-α and curcumin can inhibit the proliferation of B-NHL Raji cells apparently in vitro. Promotion of the expression of caspase 6, caspase 8, caspase 9 and induction of apoptosis might be one of the important mechanisms.
基金the National Natural Science Foundation of China (No. 30300253) and Wuhan Chenguang Science and Technology Project (No. 20065004116-25).
文摘Inhibin a is one of the candidate genes that control the ovulation in poultry. To study the genetic effects of inhibin a on apoptosis and proliferation of goose granulosa cells cultured in vitro, two RNA interference （RNAi） expression vectors, psiRNA-INHal and psiRNA-INHα2, were constructed to knock down inhibin α gene expression. After 48 h of transfection, the efficiency of these two RNAi expression vectors was examined by fluorescence microscopy. Meanwhile, inhibin protein expression levels, apoptosis indexes （AI） and proliferation indexes （PI） of granulosa cells were analyzed by flow cytometry. In addition, the supernatants were collected to assay the concentrations of estrogen （E2） and progesterone （P） by radioimmunoassay. The results showed that the expression level of inhibin a in the RNAi group were decreased 30%--40% than those in the control groups （P 〈0.05） and the apoptosis indexes and proliferation indexes in the RNAi groups were significantly higher than those in the control groups （P 〈0.05）. However, the E2 concentrations in the RNAi groups were lower than those in the control groups （P 〈0.05）. These results indicate that inhibin a has antagonistic effect on granulosa cell apoptosis.
基金Supported by grants from the National Basic Research Program of China (No. 2004CB518705)National Natural Sciences Foundation of China (No.30570908)
文摘Objective： We investigated the influence of CDK1 and CDK2 expression inhibited by cotransfection of CDK1 and CDK2 siRNA on cell cycle and apoptosis, explored the exact role of cell cycle master regulator in tumor cell apoptosis process. Methods： The siRNA targeting the CDK1 and CDK2 genes were synthesized and simultaneously cotransfected into Hela cells by lipofectamine 2000.48 or 60 h after the cotransfection, CDK1 and CDK2 protein expressions were examined by Western blot. Cell cycle distribution was analyzed by flow cytometry. Cell apoptosis was detected by the Annexin V/PI method. The changes of the transfected cell morphological under a microscope after Wright-Giemsa Staining were studied. Results： CDK1 and CDK2 protein expression was decreased at 48 or 60 h after cotransfection. The accumulation of the G2/M and S phase population in cell cycle of the cotransfected cells at 48 or 60 h after transfection was enhanced obviously compared with control. The ratio of apoptotic cell of cotransfected cells at 48 or 60 h after transfection was increased significantly compared with control. More binucleate or multinucleate ceJls among cotransfected cells were observed under the microscope. Conclusion： The decreased expression of CDK1 and CDK2 by cotransfection of CDK1 and CDK2 siRNA not only leads to tumor cell cycle arrest in S phase and G2/M phase, but also induces tumor cell apoptosis.
文摘Objective:The aim of this study was to investigate the molecular mechanism of anti-apoptotic action of survivin to the hepatoma-cellular carcinoma cell line HepG2.Methods:Design and synthesize siRNA gene sequence specifically targeting at HepG2 cell.HepG2 cells cultures were divided into five groups:blank control group,negative control group,low dose group,medium dose group and high dose group.HepG2 cells were treated respectively by pshRNA-survivin-387 of different concentrations.The apoptosis index(AI) was determined by flow cytometry(FCM).Cells were stained with rhodomine-123(Rh123) to detect changes of mitochondrial membrane potentials.The concentration of cytoplasmic cytochrome C(Cyt.C) was continuously determined by ELISA.Relative activities of caspase-9 and caspase-3 were assessed by colorimetric assay.Results:Compared with the control group,due to the function of short interference RNAs(SiRNAs) that suppresses the survivin gene expression,the apoptotic index of transfected groups were significantly higher than those of control groups(F = 13568.68,q = 110.47-327.16,P < 0.01),the apoptosis index of high concentration of transfected cells was higher than the low concentration transfected group(q = 39.63-168.22,P < 0.01).The apoptosis index of high concentrations transfected HepG2 cells was 25.54%,higher than that of blank control group,negative control group,low dose group and medium dose group(5.24%,6.61%,12.63% and 15.64%,respectively).HepG2 cells transfected with SiRNA exhibit gradually decreasing mitochondrial membrane potentials,which then lead to the releasing of Cyt C,following it were the activation of caspase-9 and caspase-3.Conclusion:Survivin performs the function of anti-apoptosis to the HepG2 cells via modulating the apoptosis of mitochondrial.HepG2 cells transfected with SiRNA survivin can significantly induce apoptosis.
