mTOR通路抑制剂GDC-0349对CCRF-CEM细胞增殖和凋亡的影响
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摘要
目的:探讨mTOR通路抑制剂GDC-0349对急性T淋巴细胞白血病细胞株CCRF-CEM细胞增殖和凋亡的影响。方法:不同浓度GDC-0349作用于CCRF-CEM细胞,采用CCK-8法检测细胞增殖;流式细胞术检测细胞凋亡率;Western blot检测凋亡相关蛋白及AKT/mTOR通路相关蛋白的表达。结果:GDC-0349作用于CCRF-CEM细胞后细胞增殖率减少且凋亡率增加(P<0.05);凋亡相关蛋白Caspase3和BCL2下调,Cleaved-Caspase3增加(P<0.05);通路相关蛋白AKT、p-AKT及p-mTOR下调(P<0.05)。结论:GDC-0349能够抑制AKT/mTOR通路活化,从而促进CCRF-CEM细胞凋亡,抑制细胞增殖。
Objective:To investigate the effect of GDC-0349,the mTOR pathway inhibitor,on proliferation and apoptosis of acute T lymphocytic leukemia cell line CCRF-CEM.Methods:Different concentrations of GDC-0349 were applied to CCRF-CEM cells.Cell proliferation was detected by CCK-8 method.Apoptosis rate was detected by flow cytometry.Expressions of apoptosis-related proteins and AKT/mTOR pathway proteins were detected by Western blot.Results:In CCRF-CEM cells with GDC-0349,the cell proliferation rate decreased and the apoptotic rate increased(P<0.05).Caspase3 and BCL2 were down-regulated(P<0.05) and Cleaved-Caspase3 was up-regulated(P<0.05).The expressions of AKT/mTOR pathway-related proteins were down-regulated,such as AKT,p-AKT and p-mTOR(P<0.05).Conclusion:GDC-0349 can inhibit the AKT/mTOR pathway,then promote cell apoptosis and inhibit cell proliferation in CCRF-CEM cells.
Objective:To investigate the effect of GDC-0349,the mTOR pathway inhibitor,on proliferation and apoptosis of acute T lymphocytic leukemia cell line CCRF-CEM.Methods:Different concentrations of GDC-0349 were applied to CCRF-CEM cells.Cell proliferation was detected by CCK-8 method.Apoptosis rate was detected by flow cytometry.Expressions of apoptosis-related proteins and AKT/mTOR pathway proteins were detected by Western blot.Results:In CCRF-CEM cells with GDC-0349,the cell proliferation rate decreased and the apoptotic rate increased(P<0.05).Caspase3 and BCL2 were down-regulated(P<0.05) and Cleaved-Caspase3 was up-regulated(P<0.05).The expressions of AKT/mTOR pathway-related proteins were down-regulated,such as AKT,p-AKT and p-mTOR(P<0.05).Conclusion:GDC-0349 can inhibit the AKT/mTOR pathway,then promote cell apoptosis and inhibit cell proliferation in CCRF-CEM cells.
引文
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[2] Guo W,Schubbert S,Chen JY,et al.Suppression of leukemia development caused by PTEN loss[J].Proceedings of the National Academy of Sciences of the United States of America,2011,108(4):1409-1414.
[3] Estrada AA,Shore DG,Blackwood E,et al.Pyrimidoaminotropanes as potent,selective,and efficacious small molecule kinase inhibitors of the mammalian target of rapamycin (mTOR) [J].Journal of Medicinal Chemistry,2013,56(7):3090-3101.
[4] Zhou Y,Peng Y,Tang H,et al.Autophagy induction contributes to GDC-0349 resistance in head and neck squamous cell carcinoma (HNSCC) cells[J].Biochemical and Biophysical Research Communications,2016,477(2):174-180.
[5] Silva A,Yunes JA,Cardoso BA,et al.PTEN posttranslational inactivation and hyperactivation of the PI3K/Akt pathway sustain primary T cell leukemia viability[J].Journal of Clinical Investigation,2008,118(11):3762-3774.
[6] Ksionda O,Mues M,Wandler AM,et al.Comprehensive analysis of T cell leukemia signals reveals heterogeneity in the PI3 kinase-Akt pathway and limitations of PI3 kinase inhibitors as monotherapy [J].PLoS One,2018,13(5):e0193849.
[7] Imbert V,Nebout M,Mary D,et al.Co-targeting intracellular pH and essential amino acid uptake cooperates to induce cell death of T-ALL/LL cells[J].Leukemia & Lymphoma,2018,59(2):460-468.
[8] Ma W,Zhu M,Yang L,et al.Synergistic effect of TPD7 and berberine against leukemia Jurkat cell growth through regulating ephrin-B2 signaling [J].Phytotherapy Research:PTR,2017,31(9):1392-1399.
[9] Bertacchini J,Heidari N,Medianl L,et al.Targeting PI3K/AKT/mTOR network for treatment of leukemia[J].Cellular and Molecular Life Sciences:CMLS,2015,72(12):2337-2347.
[10] Kawata T,Tada K,Kobayashi M,et al.Dual inhibition of the mTORC1 and mTORC2 signaling pathways is a promising therapeutic target for adult T-cell leukemia[J].Cancer Science,2018,109(1):103-111.
[11] Nitulescu GM,Margina D,Juzenas P,et al.Akt inhibitors in cancer treatment:The long journey from drug discovery to clinical use (Review) [J].International Journal of Oncology,2016,48(3):869-885.
[12] Zhao W,Qiu Y,Kong D.Class I phosphatidylinositol 3-kinase inhibitors for cancer therapy[J].Acta Pharmaceutica Sinica B,2017,7(1):27-37.
[13] Markham A.Copanlisib:First global approval[J].Drugs,2017,77(18):2057-2062.
[14] Gazi M,Moharram SA,Marhall A,et al.The dual specificity PI3K/mTOR inhibitor PKI-587 displays efficacy against T-cell acute lymphoblastic leukemia (T-ALL) [J].Cancer Letters,2017(392):9-16.
[15] Cosentino K,Garcia-Saez AJ.Bax and bak pores:Are we closing the circle[J]?Trends in Cell Biology,2017,27(4):266-275.
[16] Eleftheriadis T,Pissas G,Liakopoulos V,et al.Cytochrome c as a potentially clinical useful marker of mitochondrial and cellular damage[J].Frontiers in Immunology,2016(7):279.
[17] Khanzadeh T,Hagh MF,Talebi M,et al.Investigation of BAX and BCL2 expression and apoptosis in a resveratrol- and prednisolone-treated human T-ALL cell line,CCRF-CEM[J].Blood Research,2018,53(1):53-60.
[18] Zadi Heydarabad M,Vatanmakanian M,Abdolalizadeh J,et al.Apoptotic effect of resveratrol on human T-ALL cell line CCRF-CEM is unlikely exerted through alteration of BAX and BCL2 promoter methylation[J].Journal of Cellular Biochemistry,2018,119(12):10033-10040.
[19] Kang S,Dong SM,Kim BR,et al.Thioridazine induces apoptosis by targeting the PI3K/Akt/mTOR pathway in cervical and endometrial cancer cells[J].Apoptosis:An International Journal on Programmed Cell Death,2012,17(9):989-997.