In this study, we reported the effect of the ATP binding site competitive inhibitor Torin1 on activated α2-macroglobulin (α2M*)-induced cell proliferation and activation of mTORC1 and mTORC2 signaling in prostate ca...In this study, we reported the effect of the ATP binding site competitive inhibitor Torin1 on activated α2-macroglobulin (α2M*)-induced cell proliferation and activation of mTORC1 and mTORC2 signaling in prostate cancer cells. Torin1 significantly inhibited α2M*-induced cellproliferation as measured by protein and DNA synthesis. Translational activity, a major cellular response in malignant cells,is coordinately regulated by the mTORC1-S6-kinaseand mTORC1-4EBP1 axes. Torin1 significantly inhibited α2M*- and insulin-induced activation of mTORC1 as determined by phosphorylation of S6-kinaseat Thr389 and 4EBP1 at Thr37/46 compared to untreated cells employing Raptor immunoprecipitates. Torin1 also significantly inhibited α2M*- and insulin-induced upregulation of p-AktT308 and p-AktS473 in prostate cancer cells. The effect was comparable to that of insulin employed as a positive control. Finally, Torin1 inhibited α2M*- and insulin-induced activation of mTORC2 kinase assayas measured by phosphorylation of Akt at Ser473 inRictor immunoprecipitates of prostate cancer cells.展开更多
目的:探讨新疆维吾尔族和汉族乳腺癌组织中哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)及其下游分子4EBP1和S6K1的表达差异。方法:选取2005-03-01-2009-09-30新疆医科大学附属肿瘤医院的维吾尔族和汉族女性浸润性乳腺...目的:探讨新疆维吾尔族和汉族乳腺癌组织中哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)及其下游分子4EBP1和S6K1的表达差异。方法:选取2005-03-01-2009-09-30新疆医科大学附属肿瘤医院的维吾尔族和汉族女性浸润性乳腺癌患者285例,使用免疫组化方法检测磷酸化mTOR及其下游分子4EBP1和S6K1的表达状况;分别选取2012-01-01-2012-06-30新疆医科大学附属肿瘤医院63例维、汉新鲜乳腺癌和癌旁组织样本,使用荧光定量RT-PCR法检测mTOR及其下游分子4EBP1和S6K1的mRNA表达状况。结果:免疫组化结果显示,维吾尔族和汉族p-mTOR在乳腺癌组织中的阳性表达率分别为77.1%和67.4%(P=0.067),在癌旁组织中的阳性表达率分别为57.6%和61.7%(P=0.485);p-4EBP1在癌组织中的阳性表达率分别为31.9%和20.0%(P=0.020),在乳腺癌旁组织中的阳性表达率分别是4.9%和12.1%(P=0.029);p-S6K1在癌组织中的阳性表达率分别为41.0%和43.3%(P=0.695),在乳腺癌旁组织中的阳性表达率分别为14.0%和22.7%,P=0.054;荧光定量RT-PCR结果显示,维、汉乳腺癌组织中mTOR mRNA相对定量结果差异有统计学意义,P=0.045;4EBP1和S6K1mRNA相对定量结果差异无统计学意义,P值分别为0.128和0.404。结论:mTOR信号通路中p-4EBP1的表达水平存在维、汉民族的差异,为研究维、汉不同民族之间乳腺癌差异及个体化治疗提供了进一步研究方向。mTOR及其下游分子mRNA表达水平与磷酸化蛋白的表达并不完全一致,提示蛋白表达的调控位点可能位于转录后水平。展开更多
文摘In this study, we reported the effect of the ATP binding site competitive inhibitor Torin1 on activated α2-macroglobulin (α2M*)-induced cell proliferation and activation of mTORC1 and mTORC2 signaling in prostate cancer cells. Torin1 significantly inhibited α2M*-induced cellproliferation as measured by protein and DNA synthesis. Translational activity, a major cellular response in malignant cells,is coordinately regulated by the mTORC1-S6-kinaseand mTORC1-4EBP1 axes. Torin1 significantly inhibited α2M*- and insulin-induced activation of mTORC1 as determined by phosphorylation of S6-kinaseat Thr389 and 4EBP1 at Thr37/46 compared to untreated cells employing Raptor immunoprecipitates. Torin1 also significantly inhibited α2M*- and insulin-induced upregulation of p-AktT308 and p-AktS473 in prostate cancer cells. The effect was comparable to that of insulin employed as a positive control. Finally, Torin1 inhibited α2M*- and insulin-induced activation of mTORC2 kinase assayas measured by phosphorylation of Akt at Ser473 inRictor immunoprecipitates of prostate cancer cells.
文摘目的:探讨新疆维吾尔族和汉族乳腺癌组织中哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)及其下游分子4EBP1和S6K1的表达差异。方法:选取2005-03-01-2009-09-30新疆医科大学附属肿瘤医院的维吾尔族和汉族女性浸润性乳腺癌患者285例,使用免疫组化方法检测磷酸化mTOR及其下游分子4EBP1和S6K1的表达状况;分别选取2012-01-01-2012-06-30新疆医科大学附属肿瘤医院63例维、汉新鲜乳腺癌和癌旁组织样本,使用荧光定量RT-PCR法检测mTOR及其下游分子4EBP1和S6K1的mRNA表达状况。结果:免疫组化结果显示,维吾尔族和汉族p-mTOR在乳腺癌组织中的阳性表达率分别为77.1%和67.4%(P=0.067),在癌旁组织中的阳性表达率分别为57.6%和61.7%(P=0.485);p-4EBP1在癌组织中的阳性表达率分别为31.9%和20.0%(P=0.020),在乳腺癌旁组织中的阳性表达率分别是4.9%和12.1%(P=0.029);p-S6K1在癌组织中的阳性表达率分别为41.0%和43.3%(P=0.695),在乳腺癌旁组织中的阳性表达率分别为14.0%和22.7%,P=0.054;荧光定量RT-PCR结果显示,维、汉乳腺癌组织中mTOR mRNA相对定量结果差异有统计学意义,P=0.045;4EBP1和S6K1mRNA相对定量结果差异无统计学意义,P值分别为0.128和0.404。结论:mTOR信号通路中p-4EBP1的表达水平存在维、汉民族的差异,为研究维、汉不同民族之间乳腺癌差异及个体化治疗提供了进一步研究方向。mTOR及其下游分子mRNA表达水平与磷酸化蛋白的表达并不完全一致,提示蛋白表达的调控位点可能位于转录后水平。
