柚皮苷通过调控AKT活化影响人肺癌H1299细胞增殖、凋亡和迁移-东南大学学报医学版

柚皮苷通过调控AKT活化影响人肺癌H1299细胞增殖、凋亡和迁移

单位：1. 吉林省延边妇幼保健院药剂科, 吉林延吉 133001;
2. 吉林大学中日联谊医院肝胆胰外科, 吉林长春 130033

分类号：R734.2

出版年·卷·期（页码）：2018·37·第五期（802-807）

摘要：

目的：探讨柚皮苷对人肺癌H1299细胞增殖、凋亡以及迁移的影响。方法：柚皮苷（10、20、40 μmol·L^-1）作用于肺癌H1299细胞后，采用四甲基偶氮唑蓝（MTT）法检测H1299细胞的细胞活力；AnnexinV联合PI双染流式细胞术检测H1299细胞的凋亡率；Western blotting检测凋亡相关蛋白Bcl-2和Bax的表达水平；Caspase-3活性检测试剂盒检测法检测H1299细胞Caspase-3的活性；transwell实验检测细胞迁移。结果：柚皮苷可显著抑制H1299细胞的增殖能力，且呈浓度和时间依赖性。柚皮苷（10、20、40 μmol·L^-1）处理人肺癌H1299细胞24 h，细胞凋亡率明显高于对照组，差异有统计学意义。同时，柚皮苷（10、20、40 μmol·L^-1）处理人肺癌H1299细胞24 h，可显著下调Bcl-2、上调Bax的表达水平，增加Caspase-3的活性。柚皮苷作用于人肺癌H1299细胞24 h，可抑制其迁移能力。柚皮苷处理24 hH1299细胞的p-AKT/AKT水平显著降低。结论：柚皮苷可抑制人肺癌细胞的增殖和迁移，促进人肺癌细胞的凋亡，其机制可能与其抑制AKT的活化有关。

Objective:To explored the effects of naringin on cell proliferation, apoptosis and migration of H1299 cells. Methods:After treated H1299 cells with different concentrations of naringin (10, 20, 40 μmol·L^-1) for 24 h, cell viability was detected by using MTT assay; the rate of cell apoptosis was determined by flow cytometry (FCM) after Annexin V-FITC/PI staining; the expression levels of Bax, Bcl-2, p-AKT and AKT protein were measured by Western blotting; Caspase-3 activity was determined by Caspase-3 activity assay kit; migration was measured by transwell assay. Results:Compared with the control group, naringin obviously inhibited the viability of H1299 cells in a dose and time dependent way. Moreover, treatment with naringin (10, 20, and 40 μmol·L^-1) for 24 h, the apoptosis rate of H1299 cells were significantly increased. We also found that the level of Bcl-2 protein was reduced whereas the level of Bax protein and the activity of Caspase-3 in H1299 cells were increased after treatment with naringin (10, 20, 40 μmol·L^-1) for 24 h. Furthermore, naringin markedly inhibited migration and AKT activity of H1299 cells. Conclusion:Naringin can inhibit cell proliferation as well as migration and promote cell apoptosis of human lung cancer and its mechanism may be related to inhibition of AKT activation.

参考文献：

[1] SIEGEL R,NAISHADHAM D,JEMAL A.Cancer statistics,2013[J].Ca Cancer J Clin,2013,63(1):10-29.
[2] RAUNGRUT P,WONGKOTSILA A,LIRDPRAPAMONGKOL K,et al.Prognostic significance of 14-3-3γ overexpression in advanced non-small cell lung cancer[J].Asian Pac J Cancer Prev,2014,15(8):3513-8.
[3] 马先红,隋新,刘洋.柚皮活性成分提取工艺的研究进展[J].吉林化工学院学报,2014,31(11):38-41.
[4] 谢仁峰,文双娥,李洋,等.柚皮苷抗炎镇痛作用的实验研究[J].湖南师范大学学报医学版,2011,8(4):5-8.
[5] LIU K,WU L,SHI X,et al.Protective effect of naringin against ankylosing spondylitis via ossification,inflammation and oxidative stress in mice[J].Exp Ther Med,2016,12(2):1153.
[6] 刘新迎,周联,梁瑞燕,等.柚皮苷对前脂肪细胞3T3-L1增殖和诱导分化的影响[J].中药新药与临床药理,2007,18(3):176-179.
[7] 杨慧慧,王秀珍.柚皮苷体外抗肿瘤活性及其机制研究[J].广东药学院学报,2016,32(6):757-761.
[8] 杨文静,王璐,孙锐.不同浓度柚皮苷对人宫颈癌HeLa细胞体外增殖及其COX-2表达影响[J].中国免疫学杂志,2016,32(8):1200-1203.
[9] 宋淑慧,胡昕,熊玉卿,等.柚皮苷对人卵巢癌SKOV3细胞环氧化酶2mRNA及蛋白表达水平的影响[J].中国临床药理学与治疗学,2013,18(3):271-276.
[10] BANJERDPONGCHAI R,WUDTIWAI B,KHAWON P.Induction of Human Hepatocellular Carcinoma HepG2 Cell Apoptosis by Naringin[J].Asian Pac J Cancer Prev 2016,17(7):3289.
[11] WESOŁOWSKA O,WIŚNIEWSKI J,SRODAPOMIANEK K,et al.Multidrug resistance reversal and apoptosis induction in human colon cancer cells by some flavonoids present in citrus plants[J].J Nat Prod,2012,75(11):1896.
[12] DEY S K,BOSE D,HAZRA A,et al.Cytotoxic activity and apoptosis-inducing potential of di-spiropyrrolidino and di-spiropyrrolizidino oxindole andrographolide derivatives[J].Plos One,2013,8(3):e58055.
[13] LAVRIK I N,GOLKS A,KRAMMER P H.Caspases:pharmacological manipulation of cell death[J].J Clin Invest,2005,115(10):2665.
[14] 熊莺,王广发,张俊艳,等.柚皮苷抑制高糖诱导的脐静脉内皮细胞与单核细胞的粘附作用[J].南方医科大学学报,2010,30(2):321-325.
[15] 王巧能.枳实中黄酮类化合物的提取、分离、鉴定与生物活性研究[D].杭州:浙江工商大学,2008.
[16] 李秀娟,周志钦.柑桔柚皮苷抗癌活性研究进展[J].中国果业信息,2011,28(1):27-30.
[17] KANNO S I,AI S,HIRATA R,et al.Effects of naringin on cytosine arabinoside (Ara-C)-induced cytotoxicity and apoptosis in P388 cells[J].Life Sci,2003,75(3):353-365.
[18] JAGETIA G C,REDDY T K.Modulation of radiation-induced alteration in the antioxidant status of mice by naringin[J].Life Sci,2005,77(7):780-794.
[19] CAMARGO C A,GOMES-MARCONDES M C,WUTZKI N C,et al.Naringin inhibits tumor growth and reduces interleukin-6 and tumor necrosis factor α levels in rats with Walker 256 carcinosarcoma[J].Anticancer Res,2012,32(1):129.
[20] HATA A N,ENGELMAN J A,FABER A C.The BCL-2 family:key mediators of the apoptotic response to targeted anti-cancer therapeutics[J].Cancer Discovery,2015,5(5):475.
[21] 赵彦超,顾耘.细胞凋亡通路研究进展[J].现代医学,2013,41(4):285-288.
[22] SHI H,JI Y,ZHANG D,et al.MiR-135a inhibits migration and invasion and regulates EMT-related marker genes by targeting KLF8 in lung cancer cells[J].Biochem Biophys Res Commun,2015,465(1):125-130.
[23] 王京,黄烨清,许斌,等.长链非编码RNA RP11-191L9.4促进前列腺癌PC-3细胞的迁移和侵袭[J].东南大学学报(医学版),2016,35(3):305-310.
[24] 贾超,王洁,邵根宝,等.LSD1和KDM5家族在卵巢癌细胞中的表达及LSD1对细胞增殖迁移的影响[J].东南大学学报(医学版),2016,35(3):350-354.
[25] TOKER A,YOELILERNER M.Akt signaling and cancer:surviving but not moving on[J].Cance Res,2006,66(8):3963-3966.
[26] 苗丽君,王静,吴秋歌,等.非小细胞肺癌中AKT活化及与抗凋亡蛋白Survivin、Bcl-2表达的关系[J].天津医药,2007,35(4):241-243.

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