Objective： To investigate the apoptosis induction by arsenic trioxide （As2O3） in Raji cells and its correlation with cell cycle arrest and expression of the Survivin gene. Methods： After Raji cells were treated wi...Objective： To investigate the apoptosis induction by arsenic trioxide （As2O3） in Raji cells and its correlation with cell cycle arrest and expression of the Survivin gene. Methods： After Raji cells were treated with As2O3 in different concen- trations （1, 2, 4 and 8 pM）, for 24, 48 and 72 h, respectively, and cell proliferation was tested by MTT assay. Apoptosis was observed with electron microscope and DNA electrophoresis. The distribution of cell cycles and cell apoptosis were detected by flow cytometry. Expression of the Survivin gene was determined by real-time quantitative RT-PCR. Results： As2O3 （1-8 μM） inhibited Raji cells growth effectively in a dose- and time-dependent manner. As2O3 at 2-8μM could induce cell apoptosis and cell cycle arrest. However, As2O3（1 μM） inhibited Raji proliferation only by cell cycle arrest, without any symptoms of cell apoptosis. At the same time, Survivin gene expression was down-regulated after the treatment. Conclusion： As2O3 could induce substantial proliferation inhibition, cell cycle arrest and apoptosis in Raji cell. Cell cycle arrest might be a reason why apoptosis occurs. As2O3 can markedly down-regulate expression of the Survivin gene in a dose- and timedependent manner. The down-regulated Survivin gene might be leading to cell apoptosis by As2O3.展开更多
This study constructed siRNA recombinant expression vector targeting survivin gene and observe the apoptosis induction effect of it in human colon cancer cells, siRNA recombinant expression vector targeting survivin g...This study constructed siRNA recombinant expression vector targeting survivin gene and observe the apoptosis induction effect of it in human colon cancer cells, siRNA recombinant expression vector targeting survivin gene was constructed and transfected into human colon cancer cells. The effect of siRNA recombinant expression vector was detected by RT-PCR, Western blot, MTT reduction assay and flow cytometry. It was confirmed by restriction endonuclease and sequence analysis that siRNA recombinant expression vector targeting survivin gene was constructed successfully. Inhibition rate of survivin siRNA at mRNA and protein levels was 36.33% and 44.65% respectively. Growth of cancer cells was inhibited and the apoptosis rate was （17.24±2.13）%. The siRNA recombinant expression vector targeting survivin gene has been constructed successfully. It not only can inhibit the expression of survivin gene, but also can induce apoptosis in human colon cancer cells remarkably.展开更多
Objective:To explore the influence of extract of Ginkgo biloba（EGB） on the proliferation, apoptosis of ACC-2 cell and Survivin gene expression in adenoid cystic carcinoma（ACC） of lacrimal gland.Methods:ACC-2 cell ...Objective:To explore the influence of extract of Ginkgo biloba（EGB） on the proliferation, apoptosis of ACC-2 cell and Survivin gene expression in adenoid cystic carcinoma（ACC） of lacrimal gland.Methods:ACC-2 cell in human with ACC of lacrimal gland was in vitro cultured. MTT method was used for cell proliferation detection.Annexin V/PI double-staining flow cytometer was used to detect cell apoptosis and cell cycle.Survivin gene expression was analyzed by RT-PCR and Western blotting.Results:EGB had inhibitory effect on the proliferation of ACC-2 cell with significant dose-effect relationship,and there was statistical difference when compared with the control group（P【0.01）.The inhibitory concentration 50%(IC<sub>50</sub>) is 88 mg/L. The flow cytometer test indicated that CGB can gradually increase ACC-2 cell in G<sub>0</sub>-G<sub>1</sub> stage and decrease it in G<sub>2</sub>-M and S stage.With the increase of dose,the apoptosis rate of ACC-2 cell was obviously increased（P【0.05 or P【0.01）.EGB had certain inhibitor)’ effect on Survivin gene expression of ACC-2 cell,and Survivin gene expression was decreased with the increasing of the EGB concentration（P【0.01）.Conclusions:EGB can effectively inhibit Survivin gene expression of ACC-2 cell in human with ACC of lacrimal gland,induce the apoptosis of ACC-2 cell and inhibit tumor cell proliferation.展开更多
Survivin gene may be a good target for cancer gene therapy because it is over expressed in a variety of human tumors including human hepatocellular carcinoma but not in differen- tiated adult tissues. To explore the e...Survivin gene may be a good target for cancer gene therapy because it is over expressed in a variety of human tumors including human hepatocellular carcinoma but not in differen- tiated adult tissues. To explore the effects of the siRNA of survivin gene inducing apoptosis in human hepatocellular cancer cells, three siRNAs cpusiRNA1, cpusiRNA2 and cpusiRNA3 were designed and transferred into human hepatocellular carcinoma cell line HepG2 (HepG2) by lipofection. MTT test showed that the growth of HepG2 decreased when it was transfected with 25nM, 50nM, 100nM, 150nM, 200nM, 400nM siRNA respectively after 48 hours. And the change of mRNA and protein of survivin gene and p53 gene had been detected by RT-PCR and Western blot. Cells presented an increase in apoptosis index was assayed by flow cytometry. Small interfering RNA can exert a knockdown of survivin gene expression and up regulation of p53 gene to induce apoptosis and to inhibit the growth of HepG2.展开更多
基金a grand from the Educational Committee Scientific Research Foundation of Yunnan Province. (No. 06z095c).
文摘Objective： To investigate the apoptosis induction by arsenic trioxide （As2O3） in Raji cells and its correlation with cell cycle arrest and expression of the Survivin gene. Methods： After Raji cells were treated with As2O3 in different concen- trations （1, 2, 4 and 8 pM）, for 24, 48 and 72 h, respectively, and cell proliferation was tested by MTT assay. Apoptosis was observed with electron microscope and DNA electrophoresis. The distribution of cell cycles and cell apoptosis were detected by flow cytometry. Expression of the Survivin gene was determined by real-time quantitative RT-PCR. Results： As2O3 （1-8 μM） inhibited Raji cells growth effectively in a dose- and time-dependent manner. As2O3 at 2-8μM could induce cell apoptosis and cell cycle arrest. However, As2O3（1 μM） inhibited Raji proliferation only by cell cycle arrest, without any symptoms of cell apoptosis. At the same time, Survivin gene expression was down-regulated after the treatment. Conclusion： As2O3 could induce substantial proliferation inhibition, cell cycle arrest and apoptosis in Raji cell. Cell cycle arrest might be a reason why apoptosis occurs. As2O3 can markedly down-regulate expression of the Survivin gene in a dose- and timedependent manner. The down-regulated Survivin gene might be leading to cell apoptosis by As2O3.
基金supported by grants from National High Technology Research and Development Program of China (No. 2001AA218051)Educational Commission of Hubei Province of China (No. 2005A304B09).
文摘This study constructed siRNA recombinant expression vector targeting survivin gene and observe the apoptosis induction effect of it in human colon cancer cells, siRNA recombinant expression vector targeting survivin gene was constructed and transfected into human colon cancer cells. The effect of siRNA recombinant expression vector was detected by RT-PCR, Western blot, MTT reduction assay and flow cytometry. It was confirmed by restriction endonuclease and sequence analysis that siRNA recombinant expression vector targeting survivin gene was constructed successfully. Inhibition rate of survivin siRNA at mRNA and protein levels was 36.33% and 44.65% respectively. Growth of cancer cells was inhibited and the apoptosis rate was （17.24±2.13）%. The siRNA recombinant expression vector targeting survivin gene has been constructed successfully. It not only can inhibit the expression of survivin gene, but also can induce apoptosis in human colon cancer cells remarkably.
文摘Objective:To explore the influence of extract of Ginkgo biloba（EGB） on the proliferation, apoptosis of ACC-2 cell and Survivin gene expression in adenoid cystic carcinoma（ACC） of lacrimal gland.Methods:ACC-2 cell in human with ACC of lacrimal gland was in vitro cultured. MTT method was used for cell proliferation detection.Annexin V/PI double-staining flow cytometer was used to detect cell apoptosis and cell cycle.Survivin gene expression was analyzed by RT-PCR and Western blotting.Results:EGB had inhibitory effect on the proliferation of ACC-2 cell with significant dose-effect relationship,and there was statistical difference when compared with the control group（P【0.01）.The inhibitory concentration 50%(IC<sub>50</sub>) is 88 mg/L. The flow cytometer test indicated that CGB can gradually increase ACC-2 cell in G<sub>0</sub>-G<sub>1</sub> stage and decrease it in G<sub>2</sub>-M and S stage.With the increase of dose,the apoptosis rate of ACC-2 cell was obviously increased（P【0.05 or P【0.01）.EGB had certain inhibitor)’ effect on Survivin gene expression of ACC-2 cell,and Survivin gene expression was decreased with the increasing of the EGB concentration（P【0.01）.Conclusions:EGB can effectively inhibit Survivin gene expression of ACC-2 cell in human with ACC of lacrimal gland,induce the apoptosis of ACC-2 cell and inhibit tumor cell proliferation.
文摘Survivin gene may be a good target for cancer gene therapy because it is over expressed in a variety of human tumors including human hepatocellular carcinoma but not in differen- tiated adult tissues. To explore the effects of the siRNA of survivin gene inducing apoptosis in human hepatocellular cancer cells, three siRNAs cpusiRNA1, cpusiRNA2 and cpusiRNA3 were designed and transferred into human hepatocellular carcinoma cell line HepG2 (HepG2) by lipofection. MTT test showed that the growth of HepG2 decreased when it was transfected with 25nM, 50nM, 100nM, 150nM, 200nM, 400nM siRNA respectively after 48 hours. And the change of mRNA and protein of survivin gene and p53 gene had been detected by RT-PCR and Western blot. Cells presented an increase in apoptosis index was assayed by flow cytometry. Small interfering RNA can exert a knockdown of survivin gene expression and up regulation of p53 gene to induce apoptosis and to inhibit the growth of HepG2.
基金国家自然科学基金(No.30471599)国家重点基础研究发展(973)计划资助项目(No.2005CB724300-G)+3 种基金上海科技发展基金(No.03ZR14057)Supported by the National Natural Science Foundation of China(No.30471599)the Major State Basic Research Development(973)Program of China(No.2005CB724300-G)(03DZ14025)the Science and Technology Development Program of Shanghai(No.03ZR14057)
