鱼藤素抑制MCF7乳腺癌细胞株增殖及其机制的研究
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摘要
背景和目的:鱼藤素可抑制多种肿瘤细胞的增殖并诱导其凋亡,而且与它能抑制Akt的磷酸化相关,因此它的抗肿瘤机制可能与磷脂酰肌醇-3激酶/蛋白激酶B(phosphatidylinositol 3-kinase/protein knase B,PI3K/Akt)信号通路有关。本研究以人乳腺癌细胞株MCF7为对象,观察鱼藤素对MCF7细胞增殖的影响和能否诱导其凋亡,并研究鱼藤素调控PI3K/Akt信号通路的机制,进一步阐明它的抗肿瘤机制。
方法:
1.CCK8法检测0、1、5、10、15和20μmol/L鱼藤素作用24、48、72小时后MCF7细胞的增殖抑制率。
2.流式细胞仪Annexin V/PI双染法检测0、1、5、10、15和20μmol/L鱼藤素作用6小时后MCF7细胞的凋亡率。
3.透射电镜观察0、1、5、10、15和20μmol/L鱼藤素作用6小时后MCF7细胞的形态。
4.western blot法检测0、1和5μmol/L鱼藤素作用6小时后MCF7细胞内PI3K/Akt通路相关分子:pPTEN(Ser380)、pPDK1(Ser241)、总Akt、pAkt(Thr308)、pGSK-3β(Ser9)和pc-Raf(Ser259)的蛋白量表达。
结果:
1.鱼藤素对MCF7细胞增殖的影响:1、5、10、15和20μmol/L鱼藤素作用24小时后MCF7细胞的增殖抑制率分别为(0.77+0.32)%、(6.87+0.50)%、(11.97±1.50)%、(15.87+1.15)%和(18.23±1.80)%,鱼藤素1μmol/L组与对照组比较差异无统计学意义(p=0.408);其余各组与对照组比较差异均有显著的统计学意义(p＜0.001),抑制率随着浓度的增高而增加,各组间两两比较差异有统计学意义(p＜0.05)。48小时和72小时也出现了相似的差别。
同一浓度下,5、10、15和20μmol/L鱼藤素作用24、48和72小时后MCF7细胞的增殖抑制率均随着时间的延长而增加,各组间两两比较差异有显著的统计??学意义(p＜0.01);而1μmol/L组鱼藤素作用24、48和72小时后MCF7细胞的增殖抑制率分别为(0.77±0.32)%、(1.00±0.36)%和(0.10±0.95)%,组间比较差异无统计学意义(p=0.257)。
72小时内,5、10、15和20μmol/L鱼藤素对MCF7细胞的增殖呈抑制作用。并且随着浓度升高和时间延长其抑制作用愈加明显;1μmol/L鱼藤素对MCF7细胞的增殖无明显影响。
2.鱼藤素对MCF7细胞凋亡的诱导作用:
5、10、15和20μmol/L鱼藤素作用6小时后MCF7细胞的总凋亡率分别为(11.76±1.42)%、(15.73±0.84)%、(24.10±1.30)%和(29.83±1.66)%,各组与对照组比较差异均有显著的统计学意义(p＜0.001),随着浓度的增高其总凋亡率增加,各组间两两比较差异有显著的统计学意义(p＜0.01)。早期凋亡率和晚期凋亡率出现类似的趋势。
1μmol/L鱼藤素作用6小时后MCF7细胞的早期、晚期和总凋亡率分别为(1.47±0.66)%、(2.73±0.45)%和(4.20±0.61)%,与对照组比较P值依次为0.799、0.623和0.512,差异均无统计学意义(p＞0.05)。
5、10、15和20μmol/L鱼藤素能诱导MCF7细胞凋亡,凋亡诱导作用随着浓度的升高而愈加明显,并且透射电镜下可观察到典型的凋亡细胞形态。而1μmol/L鱼藤素对MCF7细胞的凋亡诱导作用不明显。
3.鱼藤素对MCF7细胞的PI3K/Akt信号通路相关蛋白的影响:
与对照组相比,5μmol/L鱼藤素作用6小时后,MCF7细胞内pPTEN(Ser380)、pPDK1(Ser241)、pAkt(Thr308)和pGSK-3β(Ser9)蛋白的表达量明显减少,而总Akt和pc-Raf(Ser259)蛋白的表达量则无明显变化,而1μmol/L鱼藤素作用6小时后,细胞内上述所有蛋白的表达量均无明显变化。
结论:
1.鱼藤素能抑制MCF7细胞增殖。
2.鱼藤素能诱导MCF7细胞凋亡。
3.鱼藤素能下调MCF7细胞内pPTEN(Ser380)、pPDK1(Ser241)、pAkt(Thr308)和pGSK-3β(Ser9)蛋白的表达水平,而对总Akt和pc-Raf(Ser259)蛋白的表达量无明显影响。鱼藤素可能通过抑制PTEN(Ser380)和PDK1(Ser241)蛋白的磷酸化,进而抑制Akt(Thr308)的磷酸化,间接抑制其下游GSK-3β(Ser9)的磷酸化,最终诱导MCF7细胞凋亡和抑制其增殖。
Objective:
Deguelin inhibits proliferation and induces apoptosis in a variety of cancer cells, which correlates with inhibiting the phosphorylation of protein kinase B/Akt. Deguelin's anti-cancer mechanism may be related to the phosphatidylinositol-3 kinase /protein kinase B(PI3K/Akt)signaling pathway.In this study,the effects of deguelin on proliferation and apoptosis in human breast cancer cell line MCF7 were observed. The mechanism of deguelin regulation of the PI3K/Akt signaling pathway was also explored.
Methods:
After treatment with 0,1,5,10,15 and 20μmol/L deguelin for 24,48 and 72 hours,the proliferation inhibition rate of MCF7 cells was measured by the CCK8 assay.
After treatment with 0,1,5,10,15 and 20μmol/L deguelin for 6 hours,the apoptotic rate of MCF7 cells was detected with Annexin V/PI double staining by flow cytometry and the morphologic change of MCF7 cells was observed using the transmission electron microscopy.
After treatment respectively with 0,1 and 5μmol/L deguelin for 6 hours,the total protein was extracted from MCF7 cells,then the levels of phosphorylated PTEN(Ser380),phosphorylated PDKl(Ser241),total Akt,phosphorylated Akt(Thr308),phosphorylated GSK-3p(Ser9)and phosphorylated c-Raf(Ser259) protein expression were examined by western blot analysis.
Results:
Effects of deguelin on the proliferation of MCF7 cells
After treatment with 1,5,10,15 and 20umol/L deguelin for 24 hours,the proliferation inhibition rate of MCF7 cells was(0.77±0.32)%,(6.87±0.50)%, (11.97±1.50)%,(15.87±1.15)%and(18.23±1.80)%,respectively.There was no statistical difference(p=0.408)between the lumol/L treatment group and the control group.However,there was a significant difference(p＜0.001)in the proliferation inhibition rate in each of the 5,10,15 and 20μmol/L groups compared with the control group.Furthermore,the proliferation inhibition rate increased with increasing concentration,and there were statistical differences(p＜0.05)among the 5,10,15 and 20umol/L groups.Similar differences were observed after 48 and 72 hours of deguelin.Again,there was no statistical difference in the lumol/L group but significant differences were found in each of the 5,10,15 and 20μmol/L groups versus control as well as among these groups.
At 24,48 and 72 hours,the proliferation inhibition rate increased with the longer treatment time in the 5,10,15 and 20umol/L groups,and there were statistically significant differences(p＜0.01).However,1μmol/L deguelin inhibited the proliferation of MCF7 cells by(0.77±0.32)%、(1.00±0.36)%and(0.10±0.95)%at 24, 48 and 72 hours,and there was no statistical difference among these data(p=0.257).
In short,5,10,15 and 20umol / L deguelin inhibited the proliferation of MCF7 cells at 24,48 and 72 hours.The inhibitory effect was more marked with increasing concentration and longer duration of treatment.However,1μmol/L deguelin had no significant effect.
Effects of deguelin on the apoptosis of MCF7 cells
After treatment with 5,10,15 and 20μmol/L deguelin for 6 hours,the total apoptotic rate of MCF7 cells was(11.76±1.42)%,(15.73±0.84)%,(24.10±1.30)% and(29.83±1.66)%,respectively.There were significant differences(p＜0.001) between each treatment group and the control group.The total apoptotic rate increased with increasing concentration and there were significant differences among the 5,10,15 and 20μmol/L groups(p＜0.01).Similar trends were found in the rates of early apoptosis and last apoptosis.
After treatment with 1 umol/L deguelin for 6 hours,the rate of early apoptosis, last apoptosis and total apoptosis of MCF7 cells was(1.47±0.66)%、(2.73±0.45)%and (4.20±0.61)%,respectively.Compared with the control group,the p value was 0.799、0.623 and 0.512.
In summary,5,10,15 and 20μmol/L deguelin induced apoptosis of MCF7 cells, and the apoptotic effect was more apparent with increasing concentration. Furthermore,the typical apoptotic MCF7 cells were observed under the transmission electron microscopy.But 1μmol/L deguelin had no apparent effect on inducing apoptosis of MCF7 cells.
Effects of deguelin on the expression of 6 proteins involved in the PI3K/Akt signaling pathway
After treatment with 5μmol/L deguelin for 6 hours,the expression of pPTEN (Ser380),pPDK1(Ser241),pAkt(Thr308)and pGSK-3p(Ser9)proteins was significantly reduced in MCF7 cells,while there was no significant change in the expression of total Akt and pc-Raf(Ser259)proteins.However,after treatment with 1μmol/L deguelin for 6 hours,there was no apparent change in the expression of these 6 proteins.
Conclusion:
Deguelin inhibited the proliferation of MCF7 cells.
Deguelin induced apoptosis of MCF7 cells.
Deguelin down-regulated the expression of pPTEN(Ser380),pPDK1(Ser241), pAkt(Thr308)and pGSK-3β(Ser9)protein,while it had no significant effect on the expression of total Akt and pc-Raf(Ser259)protein.Deguelin probably inhibited the phosphorylation of Akt(Thr308)and indirectly inhibited the phosphorylation of GSK-3β(Ser9)via inhibition of the phosphorylation of PTEN(Ser380)and PDKl(Ser241),thus inducing apoptosis and inhibiting the proliferation of MCF7 cells.
方法:
1.CCK8法检测0、1、5、10、15和20μmol/L鱼藤素作用24、48、72小时后MCF7细胞的增殖抑制率。
2.流式细胞仪Annexin V/PI双染法检测0、1、5、10、15和20μmol/L鱼藤素作用6小时后MCF7细胞的凋亡率。
3.透射电镜观察0、1、5、10、15和20μmol/L鱼藤素作用6小时后MCF7细胞的形态。
4.western blot法检测0、1和5μmol/L鱼藤素作用6小时后MCF7细胞内PI3K/Akt通路相关分子:pPTEN(Ser380)、pPDK1(Ser241)、总Akt、pAkt(Thr308)、pGSK-3β(Ser9)和pc-Raf(Ser259)的蛋白量表达。
结果:
1.鱼藤素对MCF7细胞增殖的影响:1、5、10、15和20μmol/L鱼藤素作用24小时后MCF7细胞的增殖抑制率分别为(0.77+0.32)%、(6.87+0.50)%、(11.97±1.50)%、(15.87+1.15)%和(18.23±1.80)%,鱼藤素1μmol/L组与对照组比较差异无统计学意义(p=0.408);其余各组与对照组比较差异均有显著的统计学意义(p＜0.001),抑制率随着浓度的增高而增加,各组间两两比较差异有统计学意义(p＜0.05)。48小时和72小时也出现了相似的差别。
同一浓度下,5、10、15和20μmol/L鱼藤素作用24、48和72小时后MCF7细胞的增殖抑制率均随着时间的延长而增加,各组间两两比较差异有显著的统计??学意义(p＜0.01);而1μmol/L组鱼藤素作用24、48和72小时后MCF7细胞的增殖抑制率分别为(0.77±0.32)%、(1.00±0.36)%和(0.10±0.95)%,组间比较差异无统计学意义(p=0.257)。
72小时内,5、10、15和20μmol/L鱼藤素对MCF7细胞的增殖呈抑制作用。并且随着浓度升高和时间延长其抑制作用愈加明显;1μmol/L鱼藤素对MCF7细胞的增殖无明显影响。
2.鱼藤素对MCF7细胞凋亡的诱导作用:
5、10、15和20μmol/L鱼藤素作用6小时后MCF7细胞的总凋亡率分别为(11.76±1.42)%、(15.73±0.84)%、(24.10±1.30)%和(29.83±1.66)%,各组与对照组比较差异均有显著的统计学意义(p＜0.001),随着浓度的增高其总凋亡率增加,各组间两两比较差异有显著的统计学意义(p＜0.01)。早期凋亡率和晚期凋亡率出现类似的趋势。
1μmol/L鱼藤素作用6小时后MCF7细胞的早期、晚期和总凋亡率分别为(1.47±0.66)%、(2.73±0.45)%和(4.20±0.61)%,与对照组比较P值依次为0.799、0.623和0.512,差异均无统计学意义(p＞0.05)。
5、10、15和20μmol/L鱼藤素能诱导MCF7细胞凋亡,凋亡诱导作用随着浓度的升高而愈加明显,并且透射电镜下可观察到典型的凋亡细胞形态。而1μmol/L鱼藤素对MCF7细胞的凋亡诱导作用不明显。
3.鱼藤素对MCF7细胞的PI3K/Akt信号通路相关蛋白的影响:
与对照组相比,5μmol/L鱼藤素作用6小时后,MCF7细胞内pPTEN(Ser380)、pPDK1(Ser241)、pAkt(Thr308)和pGSK-3β(Ser9)蛋白的表达量明显减少,而总Akt和pc-Raf(Ser259)蛋白的表达量则无明显变化,而1μmol/L鱼藤素作用6小时后,细胞内上述所有蛋白的表达量均无明显变化。
结论:
1.鱼藤素能抑制MCF7细胞增殖。
2.鱼藤素能诱导MCF7细胞凋亡。
3.鱼藤素能下调MCF7细胞内pPTEN(Ser380)、pPDK1(Ser241)、pAkt(Thr308)和pGSK-3β(Ser9)蛋白的表达水平,而对总Akt和pc-Raf(Ser259)蛋白的表达量无明显影响。鱼藤素可能通过抑制PTEN(Ser380)和PDK1(Ser241)蛋白的磷酸化,进而抑制Akt(Thr308)的磷酸化,间接抑制其下游GSK-3β(Ser9)的磷酸化,最终诱导MCF7细胞凋亡和抑制其增殖。
Objective:
Deguelin inhibits proliferation and induces apoptosis in a variety of cancer cells, which correlates with inhibiting the phosphorylation of protein kinase B/Akt. Deguelin's anti-cancer mechanism may be related to the phosphatidylinositol-3 kinase /protein kinase B(PI3K/Akt)signaling pathway.In this study,the effects of deguelin on proliferation and apoptosis in human breast cancer cell line MCF7 were observed. The mechanism of deguelin regulation of the PI3K/Akt signaling pathway was also explored.
Methods:
After treatment with 0,1,5,10,15 and 20μmol/L deguelin for 24,48 and 72 hours,the proliferation inhibition rate of MCF7 cells was measured by the CCK8 assay.
After treatment with 0,1,5,10,15 and 20μmol/L deguelin for 6 hours,the apoptotic rate of MCF7 cells was detected with Annexin V/PI double staining by flow cytometry and the morphologic change of MCF7 cells was observed using the transmission electron microscopy.
After treatment respectively with 0,1 and 5μmol/L deguelin for 6 hours,the total protein was extracted from MCF7 cells,then the levels of phosphorylated PTEN(Ser380),phosphorylated PDKl(Ser241),total Akt,phosphorylated Akt(Thr308),phosphorylated GSK-3p(Ser9)and phosphorylated c-Raf(Ser259) protein expression were examined by western blot analysis.
Results:
Effects of deguelin on the proliferation of MCF7 cells
After treatment with 1,5,10,15 and 20umol/L deguelin for 24 hours,the proliferation inhibition rate of MCF7 cells was(0.77±0.32)%,(6.87±0.50)%, (11.97±1.50)%,(15.87±1.15)%and(18.23±1.80)%,respectively.There was no statistical difference(p=0.408)between the lumol/L treatment group and the control group.However,there was a significant difference(p＜0.001)in the proliferation inhibition rate in each of the 5,10,15 and 20μmol/L groups compared with the control group.Furthermore,the proliferation inhibition rate increased with increasing concentration,and there were statistical differences(p＜0.05)among the 5,10,15 and 20umol/L groups.Similar differences were observed after 48 and 72 hours of deguelin.Again,there was no statistical difference in the lumol/L group but significant differences were found in each of the 5,10,15 and 20μmol/L groups versus control as well as among these groups.
At 24,48 and 72 hours,the proliferation inhibition rate increased with the longer treatment time in the 5,10,15 and 20umol/L groups,and there were statistically significant differences(p＜0.01).However,1μmol/L deguelin inhibited the proliferation of MCF7 cells by(0.77±0.32)%、(1.00±0.36)%and(0.10±0.95)%at 24, 48 and 72 hours,and there was no statistical difference among these data(p=0.257).
In short,5,10,15 and 20umol / L deguelin inhibited the proliferation of MCF7 cells at 24,48 and 72 hours.The inhibitory effect was more marked with increasing concentration and longer duration of treatment.However,1μmol/L deguelin had no significant effect.
Effects of deguelin on the apoptosis of MCF7 cells
After treatment with 5,10,15 and 20μmol/L deguelin for 6 hours,the total apoptotic rate of MCF7 cells was(11.76±1.42)%,(15.73±0.84)%,(24.10±1.30)% and(29.83±1.66)%,respectively.There were significant differences(p＜0.001) between each treatment group and the control group.The total apoptotic rate increased with increasing concentration and there were significant differences among the 5,10,15 and 20μmol/L groups(p＜0.01).Similar trends were found in the rates of early apoptosis and last apoptosis.
After treatment with 1 umol/L deguelin for 6 hours,the rate of early apoptosis, last apoptosis and total apoptosis of MCF7 cells was(1.47±0.66)%、(2.73±0.45)%and (4.20±0.61)%,respectively.Compared with the control group,the p value was 0.799、0.623 and 0.512.
In summary,5,10,15 and 20μmol/L deguelin induced apoptosis of MCF7 cells, and the apoptotic effect was more apparent with increasing concentration. Furthermore,the typical apoptotic MCF7 cells were observed under the transmission electron microscopy.But 1μmol/L deguelin had no apparent effect on inducing apoptosis of MCF7 cells.
Effects of deguelin on the expression of 6 proteins involved in the PI3K/Akt signaling pathway
After treatment with 5μmol/L deguelin for 6 hours,the expression of pPTEN (Ser380),pPDK1(Ser241),pAkt(Thr308)and pGSK-3p(Ser9)proteins was significantly reduced in MCF7 cells,while there was no significant change in the expression of total Akt and pc-Raf(Ser259)proteins.However,after treatment with 1μmol/L deguelin for 6 hours,there was no apparent change in the expression of these 6 proteins.
Conclusion:
Deguelin inhibited the proliferation of MCF7 cells.
Deguelin induced apoptosis of MCF7 cells.
Deguelin down-regulated the expression of pPTEN(Ser380),pPDK1(Ser241), pAkt(Thr308)and pGSK-3β(Ser9)protein,while it had no significant effect on the expression of total Akt and pc-Raf(Ser259)protein.Deguelin probably inhibited the phosphorylation of Akt(Thr308)and indirectly inhibited the phosphorylation of GSK-3β(Ser9)via inhibition of the phosphorylation of PTEN(Ser380)and PDKl(Ser241),thus inducing apoptosis and inhibiting the proliferation of MCF7 cells.
引文
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[20]Chun KH,Kosmeder JW 2nd,Sun S,et al.Effects of deguelin on the phosphatidylinositol 3-kinase/Akt pathway and apoptosis in premalignant human bronchial epithelial cells[J].J Natl Cancer Inst,2003,95(4):291-302.
[21]Lee HY,Suh YA,Kosmeder JW,et al.Deguelin-induced inhibition of cyclooxygenase-2 expression in human bronchial epithelial cells[J].Clin Cancer Res,2004,10(3):1074-1079.
[22]Geeraerts B,Vanhoecke B,Vanden Berghe W,et al.Deguelin inhibits expression of IkappaBalpha protein and induces apoptosis of B-CLL cells in vitro[J].Leukemia,2007,21(8):1610-1618.
[23]Lee JH,Lee DH,Lee HS,et al.Deguelin inhibits human hepatocellular carcinoma by antiangiogenesis and apoptosis[J].Oncol Rep,2008,20(1):129-134.
[24]Yi T,Li H,Wang X,et al.Enhancement radiosensitization of breast cancer cells by deguelin[J].Cancer Biother Radiopharm,2008,23(3):355-362.
[25]Jin Q,Feng L,Behrens C,et al.Implication of AMP-activated protein kinase and Akt-regulated survivin in lung cancer chemopreventive activities of deguelin[J].Cancer Res,2007,67(24):11630-11639.
[26]Shukla A,Grisouard J,Ehemann V,et al.Analysis of signaling pathways related to cell proliferation stimulated by insulin analogs in human mammary epithelial cell lines[J].Endocr Relat Cancer,2009 Jan 19.[Epub ahead of print]
[27]Brooks SC,Locke ER,Soule HD.Estrogen receptor in a human cell line (MCF-7) from breast carcinoma[J].J Biol Chem,1973,248(17):6251-6253.
[28]Casamayor A,Morrice NA,Alessi DR.Phosphorylation of Ser-241 is essential for the activity of 3-phosphoinositide-dependent protein kinase-1:identification of five sites of phosphorylation in vivo[J].Biochem J,1999,342 (Pt 2):287-292.
[29]Alessi DR,Andjelkovic M,Caudwell B,et al.Mechanism of activation of protein kinase B by insulin and IGF-1[J].EMBO J,1996,15(23):6541-6551.
[30]Vazquez F,Ramaswamy S,Nakamura N,et al.Phosphorylation of the PTEN tail regulates protein stability and function[J].Mol Cell Biol,2000,20(14):5010-5018.
[31]Vazquez F,Grossman SR,Takahashi Y,et al.Phosphorylation of the PTEN tail acts as an inhibitory switch by preventing its recruitment into a protein complex[J].J Biol Chem,2001,276(52):48627-48630.
[32]Das S,Dixon JE,Cho W.Membrane-binding and activation mechanism of PTEN[J].Proc Natl Acad Sci U S A,2003,100(13):7491-7496.
[33]周隽,王军臣,卢婉平,等.PTEN与预后相关因子在乳腺癌组织芯片中的相关性[J].肿瘤,2007,27(9):723-726.
[34]Diehl JA,Cheng M,Roussel MF,et al.Glycogen synthase kinase-3beta regulates cyclin D1 proteolysis and subcellular localization[J].Genes Dev,1998,12(22):3499-3511.
[35]Farago M,Dominguez I,Landesman-Bollag E,et al.Kinase-inactive glycogen synthase kinase 3beta promotes Wnt signaling and mammary tumorigenesis[J].Cancer Res,2005,65(13):5792-5801.
[36]Wang Y,Lam JB,Lam KS,et al.Adiponectin modulates the glycogen synthase kinase-3beta/beta-catenin signaling pathway and attenuates mammary tumorigenesis ofMDA-MB-231 cells in nude mice[J].Cancer Res,2006,66(23):11462-11470.
[37]Dong J,Peng J,Zhang H,et al.Role of glycogen synthase kinase 3beta in rapamycin-mediated cell cycle regulation and chemosensitivity[J].Cancer Res,2005,65(5):1961-1972.
[38]Ding Q,He X,Hsu JM,et al.Degradation of Mcl-1 by beta-TrCP mediates glycogen synthase kinase 3-induced tumor suppression and chemosensitization[J].Mol Cell Biol,2007,27(11):4006-4017.
[39]Cross DA,Alessi DR,Cohen P,et al.Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B[J].Nature,1995,378(6559):785-789.
[40]Zimmermann S,Moelling K.Phosphorylation and regulation of Raf by Akt (protein kinase B)[J].Science,1999,286(5445):1741-1744.
[1]Udeani GO,Gerhauser C,Thomas CF,et al.Cancer chemopreventive activity mediated by deguelin,a naturally occurring rotenoid[J].Cancer Res,1997,57(16):3424-3428.
[2]Gills JJ,Kosmeder J 2nd,Moon RC,et al.Effect of deguelin on UVB-induced skin carcinogenesis[J].J Chemother,2005,17(3):297-301.
[3]Murillo G,Kosmeder JW 2nd,Pezzuto JM,et al.Deguelin suppresses the formation of carcinogen-induced aberrant crypt foci in the colon of CF-1 mice[J].Int J Cancer,2003,104(1):7-11.
[4]Yan Y,Wang Y,Tan Q,Lubet RA,et al.Efficacy of deguelin and silibinin on benzo(a)pyrene-induced lung tumorigenesis in A/J mice[J].Neoplasia,2005,7(12):1053-1057.
[5]Dell'Eva R,Ambrosini C,Minghelli S,et al.The Akt inhibitor deguelin,is an angiopreventive agent also acting on the NF-kappaB pathway[J].Carcinogenesis,2007,28(2):404-413.
[6]Oh SH,Woo JK,Yazici YD,et al.Structural basis for depletion of heat shock protein 90 client proteins by deguelin[J].J Natl Cancer Inst,2007,99(12):949-961.
[7]Chun KH,Kosmeder JW 2nd,Sun S,et al.Effects of deguelin on the phosphatidylinositol 3-kinase/Akt pathway and apoptosis in premalignant human bronchial epithelial cells[J].J Natl Cancer Inst,2003,95(4):291-302.
[8]Mehta RG,Pezzuto JM.Discovery of cancer preventive agents from natural products:from plants to prevention[J].Curr Oncol Rep,2002,4(6):478-86.
[9]Kinghorn AD,Su BN,Jang DS,et al.Natural inhibitors of carcinogenesis.Planta Med,2004,70(8):691-705.
[10]Murillo G,Salti GI,Kosmeder JW 2nd,et al.Deguelin inhibits the growth of colon cancer cells through the induction of apoptosis and cell cycle arrest[J].Eur J Cancer,2002,38(18):2446-2454.
[11]Nair AS,Shishodia S,Ahn KS,et al.Deguelin,an Akt inhibitor,suppresses IkappaBalpha kinase activation leading to suppression of NF-kappaB-regulated gene expression,potentiation of apoptosis,and inhibition of cellular invasion[J].J Immunol,2006,77(8):5612-5622.
[12]Bortul R,Tazzari PL,Billi AM,et al.Deguelin,A PI3K/AKT inhibitor,enhances chemosensitivity of leukaemia cells with an active PI3K/AKT pathway[J].Br J Haematol,2005,129(5):677-686.
[13]Yi T,Li H,Wang X,et al.Enhancement radiosensitization of breast cancer cells by deguelin[J].Cancer Biother Radiopharm,2008,23(3):355-362.
[14]Kim WY,Oh SH,Woo JK,et al.Targeting heat shock protein 90 overrides the resistance of lung cancer cells by blocking radiation-induced stabilization of hypoxia-inducible factor-1 alpha[J].Cancer Res,2009 Feb 15;69(4):1624-32.
[15]Geeraerts B,Vanhoecke B,Vanden Berghe W,et al.Deguelin inhibits expression of IkappaBalpha protein and induces apoptosis of B-CLL cells in vitro [J].Leukemia,2007,21(8):1610-1618.
[16]Lee HY,Suh YA,Kosmeder JW,et al.Deguelin-induced inhibition of cyclooxygenase-2 expression in human bronchial epithelial cells[J].Clin Cancer Res.2004,10(3):1074-1079.
[17]刘红利,陈燕,崔国惠,等.鱼藤素对淋巴瘤Daudi细胞株细胞增殖细胞凋亡的影响及其机制[J].中华肿瘤杂志,2007,29(3):176-180.
[18]Lee JH,Lee DH,Lee HS,et al.Deguelin inhibits human hepatocellular carcinoma by antiangiogenesis and apoptosis[J].Oncol Rep,2008,20(1):129-134.
[19]Oh SH,Woo JK,Jin Q,et al.Identification of novel antiangiogenic anticancer activities of deguelin targeting hypoxia-inducible factor-1 alpha[J].Int J Cancer,2008,122(1):5-14.
[20]Jin Q,Feng L,Behrens C,et al.Implication of AMP-activated protein kinase and Akt-regulated survivin in lung cancer chemopreventive activities of deguelin [J].Cancer Res,2007,67(24):11630-11639.
[21]Chen WH,Chen Y,Cui GH.Deguelin inhibits expression of IkappaBalpha protein in Raji and U937 cells[J].Acta Pharmacol Sin,2006,27(4):485-490.
[22]吴秋玲,陈燕,陈卫华,等.鱼藤素对K562细胞nup98表达的影响[J].中国 实验血液学杂志,2007,15(1):25-28.
[23]Shu W,Chen Y,Wu Q,et al.Deguelin represses both the expression of nucleophosmin and some nucleoporins:Nup88 and Nup214 in Jurkat cells[J].Biol Pharm Bull,2008,31(1):27-32.
[24]陈燕,刘红利,崔国惠,等.鱼藤素对U937细胞细胞周期及核孔蛋白Nup98和Nup88的调控作用[J].中华血液学杂志,2007,28(2):115-118.
[2]Kirkegaard T,Witton CJ,McGlynn LM,et al.AKT activation predicts outcome in breast cancer patients treated with tamoxifen[J].J Pathol,2005,207(2):139-146.
[3]Tokunaga E,Kataoka A,Kimura Y,et al.The association between Akt activation and resistance to hormone therapy in metastatic breast cancer[J].Eur J Cancer,2006,42(5):629-635.
[4]Clark AS,West K,Streicher S,et al.Constitutive and inducible Akt activity promotes resistance to chemotherapy,trastuzumab,or tamoxifen in breast cancer cells[J].Mol Cancer Ther,2002,1(9):707-717.
[5]Li X,Lu Y,Liang K,et al.Differential responses to doxorubicin-induced phosphorylation and activation of Akt in human breast cancer cells[J].Breast Cancer Res,2005,7(5):R589-597.
[6]Soderlund K,Perez-Tenorio G,Stal O.Activation of the phosphatidylinositol 3-kinase/Akt pathway prevents radiation-induced apoptosis in breast cancer cells[J].Int J Oncol,2005,26(1):25-32.
[7]Liang K,Jin W,Knuefermann C,et al.Targeting the phosphatidylinositol 3-kinase/Akt pathway for enhancing breast cancer cells to radiotherapy[J].Mol Cancer Ther,2003,2(4):353-360.
[8]Campone M,Levy V,Bourbouloux E,et al.Safety and pharmacokinetics of paclitaxel and the oral mTOR inhibitor everolimus in advanced solid tumours[J].Br J Cancer,2009,100(2):315-321.
[9]Awada A,Cardoso F,Fontaine C,et al.The oral mTOR inhibitor RAD001 (everolimus) in combination with letrozole in patients with advanced breast cancer:results of a phase Ⅰ study with pharmacokinetics[J].Eur J Cancer,2008,44(1):84-91.
[10]Udeani GO,Gerhauser C,Thomas CF,et al.Cancer chemopreventive activity mediated by deguelin,a naturally occurring rotenoid[J].Cancer Res,1997,57(16):3424-3428.
[11]Murillo G,Kosmeder JW 2nd,Pezzuto JM,et al.Deguelin suppresses the formation of carcinogen-induced aberrant crypt foci in the colon of CF-1 mice[J].Int J Cancer,2003,104(1):7-11.
[12]Yan Y,Wang Y,Tan Q,et al.Efficacy of deguelin and silibinin on benzo(a)pyrene-induced lung tumorigenesis in A/J mice[J].Neoplasia,2005,7(12):1053-1057.
[13]Dell'Eva R,Ambrosini C,Minghelli S,et al.The Akt inhibitor deguelin,is an angiopreventive agent also acting on the NF-kappaB pathway[J].Carcinogenesis,2007,28(2):404-413.
[14]Oh SH,Woo JK,Yazici YD,et al.Structural basis for depletion of heat shock protein 90 client proteins by deguelin[J].J Natl Cancer Inst,2007,99(12):949-961.
[15]Gills JJ,Kosmeder J 2nd,Moon RC,et al.Effect of deguelin on UVB-induced skin carcinogenesis[J].J Chemother,2005,17(3):297-301.
[16]Chun KH,Kosmeder JW 2nd,Sun S,et al.Effects of deguelin on the phosphatidylinositol 3-kinase/Akt pathway and apoptosis in premalignant human bronchial epithelial cells[J].J Natl Cancer Inst,2003,95(4):291-302.
[17]Murillo G,Salti GI,Kosmeder JW 2nd,et al.Deguelin inhibits the growth of colon cancer cells through the induction of apoptosis and cell cycle arrest[J].Eur J Cancer,2002,38(18):2446-2454.
[18]Nair AS,Shishodia S,Ahn KS,et al.Deguelin,an Akt inhibitor,suppresses IkappaBalpha kinase activation leading to suppression of NF-kappaB-regulated gene expression,potentiation of apoptosis,and inhibition of cellular invasion[J].J Immunol,2006,177(8):5612-5622.
[19]Bortul R,Tazzari PL,Billi AM,et al.Deguelin,A PI3K/AKT inhibitor,enhances chemosensitivity of leukaemia cells with an active PI3K/AKT pathway[J].Br J Haematol,2005,129(5):677-686.
[20]Chun KH,Kosmeder JW 2nd,Sun S,et al.Effects of deguelin on the phosphatidylinositol 3-kinase/Akt pathway and apoptosis in premalignant human bronchial epithelial cells[J].J Natl Cancer Inst,2003,95(4):291-302.
[21]Lee HY,Suh YA,Kosmeder JW,et al.Deguelin-induced inhibition of cyclooxygenase-2 expression in human bronchial epithelial cells[J].Clin Cancer Res,2004,10(3):1074-1079.
[22]Geeraerts B,Vanhoecke B,Vanden Berghe W,et al.Deguelin inhibits expression of IkappaBalpha protein and induces apoptosis of B-CLL cells in vitro[J].Leukemia,2007,21(8):1610-1618.
[23]Lee JH,Lee DH,Lee HS,et al.Deguelin inhibits human hepatocellular carcinoma by antiangiogenesis and apoptosis[J].Oncol Rep,2008,20(1):129-134.
[24]Yi T,Li H,Wang X,et al.Enhancement radiosensitization of breast cancer cells by deguelin[J].Cancer Biother Radiopharm,2008,23(3):355-362.
[25]Jin Q,Feng L,Behrens C,et al.Implication of AMP-activated protein kinase and Akt-regulated survivin in lung cancer chemopreventive activities of deguelin[J].Cancer Res,2007,67(24):11630-11639.
[26]Shukla A,Grisouard J,Ehemann V,et al.Analysis of signaling pathways related to cell proliferation stimulated by insulin analogs in human mammary epithelial cell lines[J].Endocr Relat Cancer,2009 Jan 19.[Epub ahead of print]
[27]Brooks SC,Locke ER,Soule HD.Estrogen receptor in a human cell line (MCF-7) from breast carcinoma[J].J Biol Chem,1973,248(17):6251-6253.
[28]Casamayor A,Morrice NA,Alessi DR.Phosphorylation of Ser-241 is essential for the activity of 3-phosphoinositide-dependent protein kinase-1:identification of five sites of phosphorylation in vivo[J].Biochem J,1999,342 (Pt 2):287-292.
[29]Alessi DR,Andjelkovic M,Caudwell B,et al.Mechanism of activation of protein kinase B by insulin and IGF-1[J].EMBO J,1996,15(23):6541-6551.
[30]Vazquez F,Ramaswamy S,Nakamura N,et al.Phosphorylation of the PTEN tail regulates protein stability and function[J].Mol Cell Biol,2000,20(14):5010-5018.
[31]Vazquez F,Grossman SR,Takahashi Y,et al.Phosphorylation of the PTEN tail acts as an inhibitory switch by preventing its recruitment into a protein complex[J].J Biol Chem,2001,276(52):48627-48630.
[32]Das S,Dixon JE,Cho W.Membrane-binding and activation mechanism of PTEN[J].Proc Natl Acad Sci U S A,2003,100(13):7491-7496.
[33]周隽,王军臣,卢婉平,等.PTEN与预后相关因子在乳腺癌组织芯片中的相关性[J].肿瘤,2007,27(9):723-726.
[34]Diehl JA,Cheng M,Roussel MF,et al.Glycogen synthase kinase-3beta regulates cyclin D1 proteolysis and subcellular localization[J].Genes Dev,1998,12(22):3499-3511.
[35]Farago M,Dominguez I,Landesman-Bollag E,et al.Kinase-inactive glycogen synthase kinase 3beta promotes Wnt signaling and mammary tumorigenesis[J].Cancer Res,2005,65(13):5792-5801.
[36]Wang Y,Lam JB,Lam KS,et al.Adiponectin modulates the glycogen synthase kinase-3beta/beta-catenin signaling pathway and attenuates mammary tumorigenesis ofMDA-MB-231 cells in nude mice[J].Cancer Res,2006,66(23):11462-11470.
[37]Dong J,Peng J,Zhang H,et al.Role of glycogen synthase kinase 3beta in rapamycin-mediated cell cycle regulation and chemosensitivity[J].Cancer Res,2005,65(5):1961-1972.
[38]Ding Q,He X,Hsu JM,et al.Degradation of Mcl-1 by beta-TrCP mediates glycogen synthase kinase 3-induced tumor suppression and chemosensitization[J].Mol Cell Biol,2007,27(11):4006-4017.
[39]Cross DA,Alessi DR,Cohen P,et al.Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B[J].Nature,1995,378(6559):785-789.
[40]Zimmermann S,Moelling K.Phosphorylation and regulation of Raf by Akt (protein kinase B)[J].Science,1999,286(5445):1741-1744.
[1]Udeani GO,Gerhauser C,Thomas CF,et al.Cancer chemopreventive activity mediated by deguelin,a naturally occurring rotenoid[J].Cancer Res,1997,57(16):3424-3428.
[2]Gills JJ,Kosmeder J 2nd,Moon RC,et al.Effect of deguelin on UVB-induced skin carcinogenesis[J].J Chemother,2005,17(3):297-301.
[3]Murillo G,Kosmeder JW 2nd,Pezzuto JM,et al.Deguelin suppresses the formation of carcinogen-induced aberrant crypt foci in the colon of CF-1 mice[J].Int J Cancer,2003,104(1):7-11.
[4]Yan Y,Wang Y,Tan Q,Lubet RA,et al.Efficacy of deguelin and silibinin on benzo(a)pyrene-induced lung tumorigenesis in A/J mice[J].Neoplasia,2005,7(12):1053-1057.
[5]Dell'Eva R,Ambrosini C,Minghelli S,et al.The Akt inhibitor deguelin,is an angiopreventive agent also acting on the NF-kappaB pathway[J].Carcinogenesis,2007,28(2):404-413.
[6]Oh SH,Woo JK,Yazici YD,et al.Structural basis for depletion of heat shock protein 90 client proteins by deguelin[J].J Natl Cancer Inst,2007,99(12):949-961.
[7]Chun KH,Kosmeder JW 2nd,Sun S,et al.Effects of deguelin on the phosphatidylinositol 3-kinase/Akt pathway and apoptosis in premalignant human bronchial epithelial cells[J].J Natl Cancer Inst,2003,95(4):291-302.
[8]Mehta RG,Pezzuto JM.Discovery of cancer preventive agents from natural products:from plants to prevention[J].Curr Oncol Rep,2002,4(6):478-86.
[9]Kinghorn AD,Su BN,Jang DS,et al.Natural inhibitors of carcinogenesis.Planta Med,2004,70(8):691-705.
[10]Murillo G,Salti GI,Kosmeder JW 2nd,et al.Deguelin inhibits the growth of colon cancer cells through the induction of apoptosis and cell cycle arrest[J].Eur J Cancer,2002,38(18):2446-2454.
[11]Nair AS,Shishodia S,Ahn KS,et al.Deguelin,an Akt inhibitor,suppresses IkappaBalpha kinase activation leading to suppression of NF-kappaB-regulated gene expression,potentiation of apoptosis,and inhibition of cellular invasion[J].J Immunol,2006,77(8):5612-5622.
[12]Bortul R,Tazzari PL,Billi AM,et al.Deguelin,A PI3K/AKT inhibitor,enhances chemosensitivity of leukaemia cells with an active PI3K/AKT pathway[J].Br J Haematol,2005,129(5):677-686.
[13]Yi T,Li H,Wang X,et al.Enhancement radiosensitization of breast cancer cells by deguelin[J].Cancer Biother Radiopharm,2008,23(3):355-362.
[14]Kim WY,Oh SH,Woo JK,et al.Targeting heat shock protein 90 overrides the resistance of lung cancer cells by blocking radiation-induced stabilization of hypoxia-inducible factor-1 alpha[J].Cancer Res,2009 Feb 15;69(4):1624-32.
[15]Geeraerts B,Vanhoecke B,Vanden Berghe W,et al.Deguelin inhibits expression of IkappaBalpha protein and induces apoptosis of B-CLL cells in vitro [J].Leukemia,2007,21(8):1610-1618.
[16]Lee HY,Suh YA,Kosmeder JW,et al.Deguelin-induced inhibition of cyclooxygenase-2 expression in human bronchial epithelial cells[J].Clin Cancer Res.2004,10(3):1074-1079.
[17]刘红利,陈燕,崔国惠,等.鱼藤素对淋巴瘤Daudi细胞株细胞增殖细胞凋亡的影响及其机制[J].中华肿瘤杂志,2007,29(3):176-180.
[18]Lee JH,Lee DH,Lee HS,et al.Deguelin inhibits human hepatocellular carcinoma by antiangiogenesis and apoptosis[J].Oncol Rep,2008,20(1):129-134.
[19]Oh SH,Woo JK,Jin Q,et al.Identification of novel antiangiogenic anticancer activities of deguelin targeting hypoxia-inducible factor-1 alpha[J].Int J Cancer,2008,122(1):5-14.
[20]Jin Q,Feng L,Behrens C,et al.Implication of AMP-activated protein kinase and Akt-regulated survivin in lung cancer chemopreventive activities of deguelin [J].Cancer Res,2007,67(24):11630-11639.
[21]Chen WH,Chen Y,Cui GH.Deguelin inhibits expression of IkappaBalpha protein in Raji and U937 cells[J].Acta Pharmacol Sin,2006,27(4):485-490.
[22]吴秋玲,陈燕,陈卫华,等.鱼藤素对K562细胞nup98表达的影响[J].中国 实验血液学杂志,2007,15(1):25-28.
[23]Shu W,Chen Y,Wu Q,et al.Deguelin represses both the expression of nucleophosmin and some nucleoporins:Nup88 and Nup214 in Jurkat cells[J].Biol Pharm Bull,2008,31(1):27-32.
[24]陈燕,刘红利,崔国惠,等.鱼藤素对U937细胞细胞周期及核孔蛋白Nup98和Nup88的调控作用[J].中华血液学杂志,2007,28(2):115-118.