姜黄素对口腔鳞状细胞癌Tca8113细胞增殖及凋亡的影响
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摘要
目的探讨姜黄素对口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)Tca8113细胞增殖及凋亡的影响,并研究其作用机制。方法体外培养Tca8113细胞,不同浓度姜黄素(0、25、50、100μmol/L)处理24、48h;PI3K/AKT信号通路抑制剂LY294002、激动剂胰岛素样生长因子-1(IGF-1)姜黄素、IGF-1联合姜黄素处理24、48h。分别采用MTT法和流失细胞术检测细胞增殖和凋亡情况;采用Western blot和RT-PCR法检测细胞PI3K、p-AKT及mTOR的蛋白及mRNA表达水平。结果姜黄素可显著抑制OSCC Tca8113细胞的增殖并诱导凋亡,且具有一定的剂量和时间依赖性,与对照组相比差异有统计学意义(P<0.05);同时,姜黄素下调Tca8113细胞中的PI3K、p-AKT及mTOR的蛋白及mRNA表达水平(P<0.05)。IGF-1处理促进了Tca8113细胞的增殖、降低了凋亡率,同时增加PI3K、p-AKT及mTOR的蛋白及mRNA表达水平,与对照组相比差异有统计学意义(P<0.05);与IGF-1组相比,IGF-1联合姜黄素组可降低细胞的活性、提高细胞的凋亡率,降低PI3K、p-AKT及mTOR的蛋白及mRNA表达水平(P<0.05)。结论姜黄素可抑制OSCC Tca8113细胞的增殖、诱导细胞凋亡,其作用机制可能与其抑制PI3K/AKT/mTOR信号通路的表达有关。
Objective To investigate the effect of curcumin on the proliferation and apoptosis of Tca8113 cells of oral squamous cell carcinoma(OSCC)and study its mechanisms.Methods Tca8113 cells were cultured in vitro and treated with different concentrations of curcumin(0,25,50 and 100μmol/L)for 24 and 48 hours respectively.Then Tca8113 cells were treated with the PI3K/AKT signaling pathway inhibitor LY294002,agonist insulin-like growth factor-1(IGF-1),curcumin,IGF-1 combined with curcumin for 24 and 48 hours respectively.The proliferation and apoptosis of Tca8113 cells were detected by MTT assay and loss cytometry,and the protein and mRNA expression levels of PI3K,p-AKT and mTOR were detected by Western blot and RT-PCR respectively.Results Curcumin significantly inhibited the proliferation of OSCC Tca8113 cells and induced their apoptosis in a dose-and time-dependent manner,and the difference was statistically significant when compared with the control group(P<0.05).In addition,curcumin significantly down-regulated the protein and mRNA expression level of PI3K,p-AKT and mTOR in Tca8113 cells(P<0.05).IGF-1 promoted the proliferation of Tca8113 cells,reduced the apoptosis rate and increased the protein and mRNA expression levels of PI3 K,p-AKT and mTOR,and the difference was statistically significant when compared with the control group(P<0.05).Compared with the IGF-1 group,the group of IGF-1 combined with curcumin significantly decreased the cell viability,increased the apoptosis rate,and decreased the protein and mRNA expression levels of PI3K,p-AKT and mTOR(P<0.05).Conclusion Curcumin can inhibit the proliferation of OSCC Tca8113 cells and promote their apoptosis,and the mechanism may be related to the inhibition of the PI3K/AKT/mTOR signaling pathway.
Objective To investigate the effect of curcumin on the proliferation and apoptosis of Tca8113 cells of oral squamous cell carcinoma(OSCC)and study its mechanisms.Methods Tca8113 cells were cultured in vitro and treated with different concentrations of curcumin(0,25,50 and 100μmol/L)for 24 and 48 hours respectively.Then Tca8113 cells were treated with the PI3K/AKT signaling pathway inhibitor LY294002,agonist insulin-like growth factor-1(IGF-1),curcumin,IGF-1 combined with curcumin for 24 and 48 hours respectively.The proliferation and apoptosis of Tca8113 cells were detected by MTT assay and loss cytometry,and the protein and mRNA expression levels of PI3K,p-AKT and mTOR were detected by Western blot and RT-PCR respectively.Results Curcumin significantly inhibited the proliferation of OSCC Tca8113 cells and induced their apoptosis in a dose-and time-dependent manner,and the difference was statistically significant when compared with the control group(P<0.05).In addition,curcumin significantly down-regulated the protein and mRNA expression level of PI3K,p-AKT and mTOR in Tca8113 cells(P<0.05).IGF-1 promoted the proliferation of Tca8113 cells,reduced the apoptosis rate and increased the protein and mRNA expression levels of PI3 K,p-AKT and mTOR,and the difference was statistically significant when compared with the control group(P<0.05).Compared with the IGF-1 group,the group of IGF-1 combined with curcumin significantly decreased the cell viability,increased the apoptosis rate,and decreased the protein and mRNA expression levels of PI3K,p-AKT and mTOR(P<0.05).Conclusion Curcumin can inhibit the proliferation of OSCC Tca8113 cells and promote their apoptosis,and the mechanism may be related to the inhibition of the PI3K/AKT/mTOR signaling pathway.
引文
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[24]Ke J,Ma P,Chen J,et al.LGR6promotes the progression of gastric cancer through PI3K/AKT/mTOR pathway[J].Onco Targets Ther,2018,11:3025-3033.
[25]Bi Q J,Men X J,Han R,et al.LHX6inhibits the proliferation,invasion and migration of breast cancer cells by modulating the PI3K/Akt/mTOR signaling pathway[J].Eur Rev Med Pharmacol Sci,2018,22(10):3067-3073.
[26]Lv X,Li CY,Han P,et al.MicroRNA-520a-3p inhibits cell growth and metastasis of non-small cell lung cancer through PI3K/AKT/mTOR signaling pathway[J].Eur Rev Med Pharmacol Sci,2018,22(8):2321-2327.
[27]Pontes H A,de Aquino Xavier F C,da Silva T S,et al.Metallothionein and p-Akt proteins in oral dysplasia and in oral squamous cell carcinoma:an immunohistochemical study[J].J Oral Pathol Med,2009,38(8):644-650.
[2]陈新,徐文华,周健,等.口腔鳞状细胞癌现状[J].口腔医学,2017,37(5):462-465.
[3]Blatt S,Krüger M,Ziebart T,et al.Biomarkers in diagnosis and therapy of oral squamous cell carcinoma:A review of the literature[J].J Craniomaxillofac Surg,2017,45(5).
[4]Ravindran J,Prasad S,Aggarwal B B.Curcumin and cancer cells:how many ways can curry kill tumor cells selectively[J].AAPS J,2009,11(3):495-510.
[5]Anand P,Kunnumakkara A B,Newman R A,et al.Bioavailability of curcumin:problems and promises[J].Mol Pharm,2007,4(6):807-818.
[6]梁卫江,张万岱,阎曦,等.姜黄素抑制胃腺癌SGC7901细胞增殖和表皮生长因子受体表达的研究[J].胃肠病学,2006,11(7):400-403.
[7]田慧军,王笛乐.姜黄素诱导人结肠癌细胞株SW480凋亡及其bcl-2表达相关性研究[J].肿瘤防治研究,2006,33(3):185-187.
[8]杨甫文,黄金中,林晓岚,等.姜黄素对鼻咽癌细胞增殖的抑制作用[J].中国耳鼻咽喉头颈外科,2006,13(3):201-202.
[9]赵东利,谢小卫,李明众,等.姜黄素对S180小鼠体内抗肿瘤作用的实验研究[J].西安交通大学学报(医学版),2007,28(1):70-73.
[10]Mou S,Zhou Z,He Y,et al.Curcumin inhibits cell proliferation and promotes apoptosis of laryngeal cancer cells through Bcl-2and PI3K/Akt,and by upregulating miR-15a[J].Oncol Lett,2017,14(4):4937-4942.
[11]Adeyemi B F,Kolude B.Clinical presentation of oral squamous cell carcinoma[J].Niger Postgrad Med J,2013,20(2):108-110.
[12]Koprowski S,Sokolowski K,Kunnimalaiyaan S,et al.Curcumin-mediated regulation of Notch1/hairy and enhancer of split-1/survivin:molecular targeting in cholangiocarcinoma[J].J Surg Res,2015,198(2):434-440.
[13]He Y,Yue Y,Zheng X,et al.Curcumin,inflammation,and chronic diseases:how are they linked[J].Molecules,2015,20(5):9183-9213.
[14]Abdollahi E,Momtazi A A,Johnston T P,et al.Therapeutic effects of curcumin in inflammatory and immunemediated diseases:A nature-made jack-of-all-trades[J].JCell Physiol,2018,233(2):830-848.
[15]Mirzaei H,Naseri G,Rezaee R,et al.Curcumin:A new candidate for melanoma therapy[J].Int J Cancer,2016,139(8):1683-1695.
[16]Unlu A,Nayir E,Dogukan Kalenderoglu M,et al.Curcumin(Turmeric)and cancer[J].J BUON,2016,21(5):1050-1060.
[17]Maheshwari R K,Singh A K,Gaddipati J,et al.Multiple biological activities of curcumin:a short review[J].Life Sci,2006,78(18):2081-2087.
[18]Jin H,Qiao F,Wang Y,et al.Curcumin inhibits cell proliferation and induces apoptosis of human non-small cell lung cancer cells through the upregulation of miR-192-5p and suppression of PI3K/Akt signaling pathway[J].Oncol Rep,2015,34(5):2782-2789.
[19]Yu Z,Wan Y,Liu Y,et al.Curcumin induced apoptosis via PI3K/Akt-signalling pathways in SKOV3cells[J].Pharm Biol,2016,54(10):2026-2032.
[20]谭小平,徐三平,何长华,等.姜黄素诱导人胃癌细胞SGC-7901凋亡的作用机制[J].肿瘤研究与临床,2007,19(4):225-227.
[21]Simpson D R,Mell L K,Cohen E E.Targeting the PI3K/AKT/mTOR pathway in squamous cell carcinoma of the head and neck[J].Oral Oncol,2015,51(4):291-298.
[22]Liu P,Cheng H,Roberts T M,et al.Targeting the phosphoinositide 3-kinase pathway in cancer[J].Nat Rev Drug Discov,2009,8(8):627-644.
[23]Rogers S J,Box C,Harrington K J,et al.The phosphoinositide 3-kinase signalling pathway as a therapeutic target in squamous cell carcinoma of the head and neck[J].Expert Opin Ther Targets,2005,9(4):769-790.
[24]Ke J,Ma P,Chen J,et al.LGR6promotes the progression of gastric cancer through PI3K/AKT/mTOR pathway[J].Onco Targets Ther,2018,11:3025-3033.
[25]Bi Q J,Men X J,Han R,et al.LHX6inhibits the proliferation,invasion and migration of breast cancer cells by modulating the PI3K/Akt/mTOR signaling pathway[J].Eur Rev Med Pharmacol Sci,2018,22(10):3067-3073.
[26]Lv X,Li CY,Han P,et al.MicroRNA-520a-3p inhibits cell growth and metastasis of non-small cell lung cancer through PI3K/AKT/mTOR signaling pathway[J].Eur Rev Med Pharmacol Sci,2018,22(8):2321-2327.
[27]Pontes H A,de Aquino Xavier F C,da Silva T S,et al.Metallothionein and p-Akt proteins in oral dysplasia and in oral squamous cell carcinoma:an immunohistochemical study[J].J Oral Pathol Med,2009,38(8):644-650.