二吲哚吡啶基吡咯烷通过抑制AKT-mTOR阻滞MDA-MB231乳腺癌细胞周期
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摘要
探索吡咯烷衍生物的螺旋吲哚化合物二吲哚吡啶基吡咯烷(Di-indolyl pyrrolidine,DIPRD)对人乳腺癌MDAMB231细胞周期的阻滞作用。使用CCK-8法检测DIPRD对MDA-MB231细胞的毒性剂量;使用DAPI/Ed U双染法检测MDA-MB231细胞周期阻滞作用;使用Western blot检测信号通路蛋白AKT、mTOR和凋亡相关蛋白p53、MDM2的磷酸化水平以及DNA修复酶PARP的水平。DIPRD在12. 5、25、50 mg/m L剂量依赖性抑制MDA-MB231细胞活力,下调EdU阳性细胞数量,增加G_1期并减少S/G_2期细胞数量,下调p-AKT(Ser473)、p-mTOR、p-p53、cyclin D1和CDK4水平并上调p-AKT(Thr308),p-MDM2及Cleaved-PARP水平。DIPRD可能通过AKT信号通路发挥周期阻滞作用。
To investigate the induction of cell cycle arrest of human breast cancer MDA-MB231 cells by Di-indolyl pyrrolidine( DIPRD),a pyrrolidine-derived spirooxindoles compounds. The cytotoxic effect of DIPRD on MDA-MB231 cells was detected by CCK-8 method. The cell cycle arrest of MDA-MB231 cells was detected by DAPI/EdU double-staining. Phosphorylation levels of AKT,m TOR,apoptosis-related proteins p53,MDM2,and DNA repair enzyme PARP levels were detected by Western blot. DIPRD inhibited the viability of MDA-MB231 cells by downregulating the number of EdU-positive cells,increase G_1 phase and reduce cell number in S/G_2 phase,down-regulated the p-AKT( Ser473),p-m TOR,p-p53,cyclin D1,CDK4,and the upregulated the p-AKT( Thr308),p-MDM2 and Cleaved-PARP levels were detected in a dose-dependent manner at 12. 5,25,and50 mg/m L. DIPRD may play a role in cell cycle arrest through AKT signaling pathway and induce cell apoptosis.
To investigate the induction of cell cycle arrest of human breast cancer MDA-MB231 cells by Di-indolyl pyrrolidine( DIPRD),a pyrrolidine-derived spirooxindoles compounds. The cytotoxic effect of DIPRD on MDA-MB231 cells was detected by CCK-8 method. The cell cycle arrest of MDA-MB231 cells was detected by DAPI/EdU double-staining. Phosphorylation levels of AKT,m TOR,apoptosis-related proteins p53,MDM2,and DNA repair enzyme PARP levels were detected by Western blot. DIPRD inhibited the viability of MDA-MB231 cells by downregulating the number of EdU-positive cells,increase G_1 phase and reduce cell number in S/G_2 phase,down-regulated the p-AKT( Ser473),p-m TOR,p-p53,cyclin D1,CDK4,and the upregulated the p-AKT( Thr308),p-MDM2 and Cleaved-PARP levels were detected in a dose-dependent manner at 12. 5,25,and50 mg/m L. DIPRD may play a role in cell cycle arrest through AKT signaling pathway and induce cell apoptosis.
引文
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[9]Sankaran M,Chandraprakash K,Uvarani C,et al.A convenient method of facilitating Aryl-Aryl bond-formation reactions in the synthesis of biquinoline-and quinoline-bearing chromene derivatives[J].Cheminform,2013,44(14):2858-2864.
[10]Epifano F,Genovese S,Rosati O,et al.Ytterbium triflate catalyzed synthesis ofβ-functionalized indole derivatives[J].Tetrahedr Lett,2011,52(5):568-571.
[11]Bello C,Bai J,Zambron BK,et al.Induction of cell killing and autophagy by amphiphilic pyrrolidine derivatives on human pancreatic cancer cells[J].Eur J Med Chem,2018,150:457-478.
[12]Qin S,Yang C,Huang W,et al.Sulforaphane attenuates microglia-mediated neuronal necroptosis through down-regulation of MAPK/NF-κB signaling pathways in LPS-activated BV-2 microglia[J].Pharmacol Res,2018,133:325-326.
[13]Park J,Lee YR,Shim HJ,et al.Control of tumor suppressor protein PTEN by dimethlysulfoxide in HL-60 cells[J].J Clin Oncol,2004,22(14):3156-3156.
[14]Hers I,Vincent EE,Tavara JM.Akt signalling in health and disease[J].Cell Signal,2011,23(10):1515-1527.
[15]Tuo Y,Xiang M.m TOR:a double-edged sword for diabetes[J].JLeukocyte Biol,2018,3:21-29.
[16]Cao C,Zhang HN,Yang ZL,et al.Peptide recognition,signaling and modulation of class B G protein-coupled receptors[J].Curr Opin Struc Biol,2018,51:53-60.
[17]Indio V,Astolfi A,Tarantino G,et al.Integrated molecular characterization of gastrointestinal stromal tumors(GIST)harboring the rare D842V mutation in PDGFRA gene[J].Int J Mol Sci,2018,19(3):732-747.
[18]Araldi D,Ferrari LF,Levine JD.Role of GPCR(mu-opioid)-receptor tyrosine kinase(epidermal growth factor)crosstalk in opioid-induced hyperalgesic priming(type II)[J].Pain,2018,159(5):864-875.
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[22]Li Z,Wu Y,Chen H,et al.MTOR suppresses environmental particle-induced inflammatory response in macrophages[J].JImmunol,2018,200(8):2826-2834.
[23]Liu YY,Chen MB,Cheng L,et al.microRNA-200a downregulation in human glioma leads to Gαi1 over-expression,Akt activation,and cell proliferation[J].Oncogene,2018,37(21):2890-2902.
[24]Dazert E,Hall MN.m TOR signaling in disease[J].Curr Opin Cell Biol,2011,23(6):744-755.
[25]Mi C,Ma J,Wang KS,et al.Imperatorin suppresses proliferation and angiogenesis of human colon cancer cell by targeting HIF-1αvia the m TOR/p70S6K/4E-BP1 and MAPK pathways[J].J Ethnopharmacol,2017,203:27-38.
[26]Haupt Y,Maya R,Kazaz A,et al.MDM2 promotes the rapid degradation of p53[J].Nature,1997,387(6630):296-299.
[27]Li Y,Stockton ME,Bhuiyan I,et al.MDM2 inhibition rescues neurogenic and cognitive deficits in a mouse model of fragile Xsyndrome[J].Sci Transl Med,2016,8(336):336ra61.
[28]Zhou BP,Liao Y,Xia W,et al.HER-2/neu induces p53 ubiquitination via Akt-mediated MDM2 phosphorylation[J].Nat Cell Biol,2001,3(11):973-982.
[29]Choy MK,Movassagh M,Bennett MR,et al.PKB/Akt activation inhibits p53-mediated HIF1A degradation that is independent of MDM2[J].J Cell Physiol,2010,222(3):635-639.
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