黄芪水提物抑制PI3K/Akt通路预防肺癌发生的作用及机制研究
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摘要
目的探讨黄芪水提物对肺癌的抑制作用及相关机制。方法采用MTT实验、Ed U实验以及划痕实验检测黄芪水提物对肺癌A549和H1299细胞的活性、增殖能力和迁移能力的影响,采用流式细胞仪考察黄芪水提物对肺癌细胞周期的影响,通过Western blotting实验检测PI3K/Akt通路相关蛋白表达,通过苯并芘诱导的小鼠肺癌模型观察黄芪水提物对体内肺癌的影响。结果 MTT实验结果表明,黄芪水提物作用于A549和H1299细胞48 h后,细胞的生长活性被抑制;Ed U实验结果表明,黄芪水提物能够显著抑制A549和H1299细胞的增殖能力;对细胞周期考察的实验结果表明,黄芪水提物能够将A549和H1299细胞周期阻滞在S期;划痕实验结果表明,黄芪水提物作用于A549和H1299细胞24 h和48 h后,能够显著抑制细胞的迁移能力;Western blotting实验结果表明,黄芪水提物能够下调A549和H1299细胞磷脂酰肌醇3激酶(PI3K)、磷酸化的磷酸肌醇依赖性蛋白激酶1(p-PDK1)和磷酸化的蛋白激酶B(p-Akt)蛋白表达水平;体内实验结果表明,黄芪水提物能够显著抑制小鼠肺部肿瘤数量及大小。结论黄芪水提物能够抑制肺癌,该机制可能与PI3K/Akt信号通路受阻有关。
Objective To investigate the inhibitory effect and mechanism of the water extracts of Astragali Radix on lung cancer. Methods The effects of Astragali Radix water extracts on cell viability, cell proliferation and cell migration of A549 and H1299 cells were detected by MTT assay, Ed U assay and wound healing assay, respectively. The effect of Astragali Radix water extracts on cell cycle was detected by flow cytometry. Western blotting assay was used to study the expression of related proteins in PI3 K/Akt pathway. Finally, the effect of Astragali Radix water extracts on lung cancer in vivo was observed by benzopyrene-induced lung cancer model. Results From the result of MTT assay, the activity of A549 and H1299 cells was inhibited after treated with Astragali Radixwater extracts after 48 h. In the Ed U experiment, the proliferation ability of A549 and H1299 cells was significantly inhibited by Astragali Radix water extracts in a concentration dependent manner. The study of propidium iodide staining assay result showed that the cell cycle of A549 and H1299 cells was arrested in S phase. In addition, the study of wound healing assay revealed that the migration ability of A549 and H1299 cells was significantly suppressed after 24 h and 48 h treatment by Astragali Radix water extracts. And Western blotting assay revealed that, in drug treatment group, the protein expressions of PI3K, p-PDK1, and p-Akt were down-regulated. Finally, in vivo experiment showed that Astragali Radix water extracts significantly inhibited the number and size of lung tumor nodules in mice. Conclusion It is concluded that Astragali Radix water extracts can inhibit lung cancer. And the mechanism may be related to the inhibition of PI3 K/Akt signaling pathway.
Objective To investigate the inhibitory effect and mechanism of the water extracts of Astragali Radix on lung cancer. Methods The effects of Astragali Radix water extracts on cell viability, cell proliferation and cell migration of A549 and H1299 cells were detected by MTT assay, Ed U assay and wound healing assay, respectively. The effect of Astragali Radix water extracts on cell cycle was detected by flow cytometry. Western blotting assay was used to study the expression of related proteins in PI3 K/Akt pathway. Finally, the effect of Astragali Radix water extracts on lung cancer in vivo was observed by benzopyrene-induced lung cancer model. Results From the result of MTT assay, the activity of A549 and H1299 cells was inhibited after treated with Astragali Radixwater extracts after 48 h. In the Ed U experiment, the proliferation ability of A549 and H1299 cells was significantly inhibited by Astragali Radix water extracts in a concentration dependent manner. The study of propidium iodide staining assay result showed that the cell cycle of A549 and H1299 cells was arrested in S phase. In addition, the study of wound healing assay revealed that the migration ability of A549 and H1299 cells was significantly suppressed after 24 h and 48 h treatment by Astragali Radix water extracts. And Western blotting assay revealed that, in drug treatment group, the protein expressions of PI3K, p-PDK1, and p-Akt were down-regulated. Finally, in vivo experiment showed that Astragali Radix water extracts significantly inhibited the number and size of lung tumor nodules in mice. Conclusion It is concluded that Astragali Radix water extracts can inhibit lung cancer. And the mechanism may be related to the inhibition of PI3 K/Akt signaling pathway.
引文
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[19]岳文涛,王小敏,王玥.PI3K/Akt/m TOR信号转导途径与非小细胞肺癌的关系[J].中国肺癌杂志,2009,12(4):312-315.
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[21]Fan X,Wang Y,Jiang T,et al.B-myb mediates proliferation and migration of non-small-cell lung cancer via suppressing IGFBP3[J].Int J Mol Sci,2018,doi:10.3390/ijms19051479.
[22]孙婷.乌拉坦诱导小鼠肺癌模型的研究[D].开封:河南大学,2013.
[23]顾其华,胡成平,夏莹,等.3,4-苯并芘肺内注射构建大鼠肺肿瘤模型的实验研究[J].中国肺癌杂志,2007,10(3):172-175.
[2]Migliorino M R,Santo A,Romano G,et al.Economic burden of patients affected by non-small cell lung cancer(NSCLC):The life study[J].J Cancer Res Clin Oncol,2017,143(5):1-9.
[3]Liu J,Zhao Z Z,Chen H B.Review of Astragali Radix[J].Chin Herb Med,2011,3(2):90-105.
[4]唐黎,武明花.黄芪注射液联合化疗治疗恶性肿瘤临床观察[J].中国医师杂志,2006,8(3):425-426.
[5]张莹,贾英杰,李小江,等.注射用黄芪多糖联合CIK细胞治疗中晚期气虚型非小细胞肺癌的临床观察[J].中草药,2018,49(7):1647-1651.
[6]Courtney K D,Corcoran R B,Engelman J A.The PI3K pathway as drug target in human cancer[J].J Clin Oncol,2010,28(6):1075-1083.
[7]Shi J,Zheng L,Lin Z F,et al.Study of pharmacokinetic profiles and characteristics of active components and their metabolites in rat plasma following oral administration of the water extract of Astragali Radix using UPLC-MS/MS[J].J Ethnopharmacol,2015,169:183-194.
[8]She J,Yang P,Hong Q,et al.Lung cancer in China:Challenges and interventions[J].Chest,2013,143(4):1117-1126.
[9]Ferlay J,Shin H R,Bray F,et al.Estimates of worldwide burden of cancer in 2008:GLOBOCAN 2008[J].Int J Cancer,2010,127(12):2893-2917.
[10]廉政君,张洪亮.晚期非小细胞肺癌中西医结合治疗进展[J].亚太传统医药,2011,7(2):143-144.
[11]张莹,王蕾,杜梦楠.注射用黄芪多糖治疗肿瘤的临床研究进展[J].药物评价研究,2016,39(6):1092-1094.
[12]Shahzad M,Shabbir A,Wojcikowski K,et al.The antioxidant effects of Radix Astragali(Astragalus membranaceus and related species)in protecting tissues from injury and disease[J].Curr Drug Targets,2016,17(12):1331-1340.
[13]薛晓利,张建琴,马雯,等.基于1H-NMR代谢组学的黄芪甲苷对秀丽隐杆线虫寿命的延长作用研究[J].中草药,2018,49(10):2274-2281.
[14]董竞成,董晓辉.黄芪注射液与白细胞介素2增强树突细胞抗肿瘤转移作用的比较研究[J].中国中西医结合杂志,2005,25(3):236-239.
[15]赵莲华,李清,陈咏梅,等.黄芪成分F3新制剂对人CCL-229、K562癌细胞的抑制作用[J].实用癌症杂志,2003,18(3):242-244.
[16]葛信艳,黄玮,李瑞琴,等.PI3K/Akt信号转导通路与肿瘤发生相互作用的机制研究[J].中国现代医药杂志,2017,19(7):98-101.
[17]倪琛琛,于敏,张志红.EGFR与PI3K/AKT信号通路相关蛋白在非小细胞肺癌组织中的表达及其意义[J].安徽医科大学学报,2011,46(12):1264-1266.
[18]廖东卫,王莉,张新光,等.PTEN/PI3K信号转导相关蛋白在非小细胞肺癌组织中的表达及其意义[J].癌症,2006,25(10):1238-1242.
[19]岳文涛,王小敏,王玥.PI3K/Akt/m TOR信号转导途径与非小细胞肺癌的关系[J].中国肺癌杂志,2009,12(4):312-315.
[20]Marinov M,Fischer B,Arcaro A.Targeting m TOR signaling in lung cancer[J].Crit Rev Oncol Hematol,2007,63(2):172-182.
[21]Fan X,Wang Y,Jiang T,et al.B-myb mediates proliferation and migration of non-small-cell lung cancer via suppressing IGFBP3[J].Int J Mol Sci,2018,doi:10.3390/ijms19051479.
[22]孙婷.乌拉坦诱导小鼠肺癌模型的研究[D].开封:河南大学,2013.
[23]顾其华,胡成平,夏莹,等.3,4-苯并芘肺内注射构建大鼠肺肿瘤模型的实验研究[J].中国肺癌杂志,2007,10(3):172-175.