Akt抑制剂MK-2206对肝癌细胞huh7生物学行为的影响及机制
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摘要
目的探讨MK-2206作用于肝癌细胞huh7后对其生物学行为的影响及作用机制。方法体外培养人肝癌细胞系huh7,采用不同浓度(0、2.5、5、10、20、40μmol/L)MK-2206干预huh7细胞。采用CCK-8检测细胞增殖变化;流式细胞术检测细胞凋亡的变化;蛋白免疫印迹法检测相关蛋白的表达。结果与对照组(0μmol/L)比较,MK-2206对肝癌细胞huh7的增殖活性有明显抑制作用,且呈剂量依赖性和时间依赖性(P <0.05);MK-2206诱导huh7细胞凋亡(P <0.05),显著抑制Akt的蛋白表达(P <0.05)。结论 Akt抑制剂MK-2206可通过调控PI3K/Akt信号通路来调控肝癌细胞huh7的生物学行为。
Objective To investigate the effect of MK-2206 on the growth of hepatoma cells huh7 and its mechanism of action. Methods Human hepatoma cell line huh7 was cultured in vitro, and huh7 cells were treated with different concentrations(0, 2.5, 5, 10, 20, 40 μmol/L) MK-2206. Cell proliferation was detected by CCK-8; apoptosis was detected by flow cytometry; protein expression was detected by Western blot. Results Compared with the control group(0 μmol/L), MK-2206 significantly inhibited the proliferation of hepatoma cell line huh7 in a dose-and time-dependent manner.MK-2206 had the tendency to reduce the migration ability of huh7, but there was no statistically significant difference.MK-2206 induced apoptosis of huh7 cells(P < 0.05) and significantly inhibited Akt protein expression(P < 0.05).Conclusion The Akt inhibitor MK-2206 can regulates the biological behavior of hepatoma cell huh7 by regulating the PI3 K/Akt signaling pathway.
Objective To investigate the effect of MK-2206 on the growth of hepatoma cells huh7 and its mechanism of action. Methods Human hepatoma cell line huh7 was cultured in vitro, and huh7 cells were treated with different concentrations(0, 2.5, 5, 10, 20, 40 μmol/L) MK-2206. Cell proliferation was detected by CCK-8; apoptosis was detected by flow cytometry; protein expression was detected by Western blot. Results Compared with the control group(0 μmol/L), MK-2206 significantly inhibited the proliferation of hepatoma cell line huh7 in a dose-and time-dependent manner.MK-2206 had the tendency to reduce the migration ability of huh7, but there was no statistically significant difference.MK-2206 induced apoptosis of huh7 cells(P < 0.05) and significantly inhibited Akt protein expression(P < 0.05).Conclusion The Akt inhibitor MK-2206 can regulates the biological behavior of hepatoma cell huh7 by regulating the PI3 K/Akt signaling pathway.
引文
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[14]Lu JW,Lin YM,Lai YL,et al.MK-2206 induces apoptosis of AML cells and enhances the cytotoxicity of cytarabine[J].Med Oncol,2015,32(7):206.
[15]Ji D,Zhang Z,Cheng L,et al.The combination of RAD001and MK-2206 exerts synergistic cytotoxic effects against PTEN mutant gastric cancer cells:involvement of MAPK-dependent autophagic,but not apoptotic cell death pathway[J].PLoS One,2014,9(1):e85 116.
[16]Malkomes P,Lunger I,Luetticke A,et al.Selective AKTinhibition by MK-2206 represses colorectal cancer-initiating stem cells[J].Ann Surg Oncol,2016,23(9):2849-2857.
[17]Gupta S,Argiles G,Munster PN,et al.A phaseⅠtrial of combined ridaforolimus and MK-2206 in patients with advanced malignancies[J].Clin Cancer Res,2015,21(23):5235-5244.
[18]Hu C,Dadon T,Chenna V,et al.Combined inhibition of cyclin-dependent kinases(Dinaciclib)and AKT(MK-2206)blocks pancreatic tumor growth and metastases in patient-derived xenograft models[J].Mol Cancer Ther,2015,14(7):1532-1539.
[19]Ramanathan RK,Mcdonough SL,Kennecke HF,et al.Phase 2 study of MK-2206,an allosteric inhibitor of AKT,as second-line therapy for advanced gastric and gastroesophageal junction cancer:A SWOG cooperative group trial(S1005)[J].Cancer,2015,121(13):2193-2197.
[2]Zhu RX,Seto WK,Lai CL,et al.Epidemiology of Hepatocellular Carcinoma in the Asia-Pacific Region[J].Gut Liver,2016,10(3):332-339.
[3]Cha C.Surgical therapy for hepatocellular carcinoma:formulating a rational approach[J].J Clin Gastroenterol,2013,47 Suppl:S30-S36.
[4]Forner A,Gilabert M,Bruix J,et al.Treatment of intermediate-stage hepatocellular carcinoma[J].Nat Rev Clin Oncol,2014,11(9):525-535.
[5]Mazzaferro V.Sorafenib in advanced hepatocellular carcinoma[J].N Engl J Med,2008,359(23):378-390.
[6]Cheng AL,Kang YK,Chen Z,et al.Efficacy and safety of sorafenib in patients in the Asia-Pacific region with advanced hepatocellular carcinoma:a phaseⅢrandomised,double-blind,placebo-controlled trial[J].Lancet Oncology,2009,10(1):4-5.
[7]Wang FZ,Peng-Jiao,Yang NN,et al.PF-04691502 triggers cell cycle arrest,apoptosis and inhibits the angiogenesis in hepatocellular carcinoma cells[J].Toxicol Lett,2013,220(2):150-156.
[8]Agarwal E,Chaudhuri A,Leiphrakpam PD,et al.Akt inhibitor MK-2206 promotes anti-tumor activity and cell death by modulation of AIF and Ezrin in colorectal cancer[J].BMC Cancer,2014,14:145.
[9]Ma CX,Suman V,Goetz MP,et al.A PhaseⅡtrial of neoadjuvant MK2206,an AKT inhibitor,with anastrozole in clinical stage 2 or 3 PIK3CA mutant ER positive and HER2 negative breast cancer[J].Clinical Cancer Res,2017,23(22):6823-6832.
[10]Llovet JM,Villanueva A,Lachenmayer A,et al.Advances in targeted therapies for hepatocellular carcinoma in the genomic era[J].Nat Rev Clin Oncol,2015,12(7):408-424.
[11]Zhang CZ,Wang XD,Wang HW,et al.Sorafenib inhibits liver cancer growth by decreasing mTOR,AKT,and PI3Kexpression[J].J BUON,2015,20(1):218-222.
[12]Li YC,He SM,He ZX,et al.Plumbagin induces apoptotic and autophagic cell death through inhibition of the PI3K/Akt/mTOR pathway in human non-small cell lung cancer cells[J].Cancer Lett,2014,344(2):239-259.
[13]Yang J,Pi C,Wang G.Inhibition of PI3K/Akt/mTOR pathway by apigenin induces apoptosis and autophagy in hepatocellular carcinoma cells[J].Biomed Pharmacother,2018,103:699-707.
[14]Lu JW,Lin YM,Lai YL,et al.MK-2206 induces apoptosis of AML cells and enhances the cytotoxicity of cytarabine[J].Med Oncol,2015,32(7):206.
[15]Ji D,Zhang Z,Cheng L,et al.The combination of RAD001and MK-2206 exerts synergistic cytotoxic effects against PTEN mutant gastric cancer cells:involvement of MAPK-dependent autophagic,but not apoptotic cell death pathway[J].PLoS One,2014,9(1):e85 116.
[16]Malkomes P,Lunger I,Luetticke A,et al.Selective AKTinhibition by MK-2206 represses colorectal cancer-initiating stem cells[J].Ann Surg Oncol,2016,23(9):2849-2857.
[17]Gupta S,Argiles G,Munster PN,et al.A phaseⅠtrial of combined ridaforolimus and MK-2206 in patients with advanced malignancies[J].Clin Cancer Res,2015,21(23):5235-5244.
[18]Hu C,Dadon T,Chenna V,et al.Combined inhibition of cyclin-dependent kinases(Dinaciclib)and AKT(MK-2206)blocks pancreatic tumor growth and metastases in patient-derived xenograft models[J].Mol Cancer Ther,2015,14(7):1532-1539.
[19]Ramanathan RK,Mcdonough SL,Kennecke HF,et al.Phase 2 study of MK-2206,an allosteric inhibitor of AKT,as second-line therapy for advanced gastric and gastroesophageal junction cancer:A SWOG cooperative group trial(S1005)[J].Cancer,2015,121(13):2193-2197.
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