姜黄素对人甲状腺癌B-CPAP细胞生物学行为的影响
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摘要
目的:探讨姜黄素对人甲状腺癌B-CPAP细胞生长、凋亡的影响及可能的作用机制。方法:采用含有不同浓度(10μmol/L、20μmol/L、40μmol/L)姜黄素的培养基培养人甲状腺癌B-CPAP细胞12 h、24 h、36h、48 h、72 h、96 h,应用MTT实验观察姜黄素对B-CPAP细胞生长的影响;采用含有不同浓度姜黄素的培养基培养人甲状腺癌B-CPAP细胞36 h、48 h、72 h、96 h,应用Annexin V/PI染色评价姜黄素诱导B-CPAP细胞凋亡的作用;应用蛋白印迹法检测姜黄素对ERK通路、Bax、Bcl-2表达的影响。结果:浓度为10μmol/L、20μmol/L、40μmol/L的姜黄素培养甲状腺癌B-CPAP细胞,随着培养时间延长,各浓度下细胞生长抑制率、细胞凋亡率均逐渐增高(P <0. 05);且随着浓度增加,相同时间点的细胞生长抑制率、细胞凋亡率亦逐渐增高(P <0. 05)。与空白组比较,姜黄素培养72 h后,B-CPAP细胞的H-Ras、p-Raf-B、p-ERK1/2、Bcl-2蛋白表达水平明显降低(P <0. 05),Bax蛋白表达水平水平明显升高(P <0. 05)。结论:姜黄素能够抑制人甲状腺癌B-CPAP细胞生长,诱导细胞凋亡,其机制可能与抑制H-Ras蛋白、ERK信号通路蛋白表达及上调Bax表达、下调Bcl-2表达有关。
Objective: To investigate the effect of curcumin on the growth and apoptosis of human thyroid carcinoma B-CPAP cells and its possible mechanism. Methods: The human thyroid carcinoma B-CPAP cells were cultured in medium containing different concentrations( 10 μmol/L,20 μmol/L,40 μmol/L) of curcumin for 12 h,24 h,36 h,48 h,72 h and 96 h. The effect of curcumin on the growth of B-CPAP cells was observed by MTT. The human thyroid carcinoma B-CPAP cells were cultured in medium containing different concentrations of curcumin for 36 h,48 h,72 h and 96 h. The effects of curcumin in inducing apoptosis of B-CPAP cells were evaluated with Annexin V/PI staining,and the effect of curcumin on ERK pathway,Bax and Bcl-2 expression was detected by Western blot.Results: The growth inhibition rate and apoptosis rate of human thyroid carcinoma B-CPAP cells cultured in medium containing 10 μmol/L,20 μmol/L and 40 μmol/L of curcumin increased gradually with the prolongation of culture time( P < 0. 05). With the increase of concentration,the growth inhibition rate and the apoptosis rate also increased gradually at the same time point( P < 0. 05). After 72 h of culture,the expression levels of H-Ras,p-Raf-B,p-ERK1/2 and Bcl-2 proteins in B-CPAP cells were significantly lower,while the expression level of Bax protein was significantly higher than that in the blank control group( P < 0. 05). Conclusion: Curcumin can inhibit the growth and induce apoptosis of human thyroid carcinoma B-CPAP cells. The mechanism may be related to inhibiting the expression of H-Ras protein and ERK signaling pathway,up-regulating the expression of Bax and down regulating the expression of Bcl-2.
Objective: To investigate the effect of curcumin on the growth and apoptosis of human thyroid carcinoma B-CPAP cells and its possible mechanism. Methods: The human thyroid carcinoma B-CPAP cells were cultured in medium containing different concentrations( 10 μmol/L,20 μmol/L,40 μmol/L) of curcumin for 12 h,24 h,36 h,48 h,72 h and 96 h. The effect of curcumin on the growth of B-CPAP cells was observed by MTT. The human thyroid carcinoma B-CPAP cells were cultured in medium containing different concentrations of curcumin for 36 h,48 h,72 h and 96 h. The effects of curcumin in inducing apoptosis of B-CPAP cells were evaluated with Annexin V/PI staining,and the effect of curcumin on ERK pathway,Bax and Bcl-2 expression was detected by Western blot.Results: The growth inhibition rate and apoptosis rate of human thyroid carcinoma B-CPAP cells cultured in medium containing 10 μmol/L,20 μmol/L and 40 μmol/L of curcumin increased gradually with the prolongation of culture time( P < 0. 05). With the increase of concentration,the growth inhibition rate and the apoptosis rate also increased gradually at the same time point( P < 0. 05). After 72 h of culture,the expression levels of H-Ras,p-Raf-B,p-ERK1/2 and Bcl-2 proteins in B-CPAP cells were significantly lower,while the expression level of Bax protein was significantly higher than that in the blank control group( P < 0. 05). Conclusion: Curcumin can inhibit the growth and induce apoptosis of human thyroid carcinoma B-CPAP cells. The mechanism may be related to inhibiting the expression of H-Ras protein and ERK signaling pathway,up-regulating the expression of Bax and down regulating the expression of Bcl-2.
引文
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[19]Yuko E,Katsuhiko Y,Yuichi K,et al.Green tea extract attenuates MNU-induced photoreceptor cell apoptosis via suppression of heme oxygenase-1[J].Journal of Toxicologic Pathology,2016,29(1):61-65.
[2]Li Jing,Luan Hong,Li Yan,et al.Clinical retrospective analysis of634 cases of thyroid carcinoma[J].Chinese Journal of Clinicians,2016,44(4):70-72.[李静,栾虹,李妍,等.甲状腺癌634例临床回顾性分析[J].中国临床医生杂志,2016,44(4):70-72.]
[3]Li Weifeng,Jiang Jianlan.Study on pharmacological action of curcumin[J].Chinese Journal of Clinical Pharmacology,2017,33(10):957-960.[李伟锋,蒋建兰.姜黄素药理作用的研究现状[J].中国临床药理学杂志,2017,33(10):957-960.]
[4]Xu Xinming,Chen Jian,Lu Rongzhu,et al.Study on the effect of curcumin on the apoptosis of SMMC-7721 cells in hepatocellular carcinoma[J].Journal of Southeast University(Medical Edition),2013,37(01):12-17.[徐新明,陈健,陆荣柱,等.姜黄素通过内质网应激信号通路诱导肝癌SMMC-7721细胞凋亡的作用研究[J].东南大学学报(医学版),2018,37(01):12-17.]
[5]Raman Dhivya,Jothi Ranjani,Jeyaprakash Rajendhran,et al.Enhancing the anti-gastric cancer activity of curcumin with biocompatible and p H sensitive PMMA-AA/Zn O nanoparticles[J].Materials Science&Engineering C,2018,82(05):101-107.
[6]Sun Xiaoran,Sun Jianjing,Zhang Linxi.Progress in the study of curcumin reversing multidrug resistance in tumor[J].Chinese Tumors,2016,25(4):272-277.[孙晓冉,孙剑经,张林西.姜黄素逆转肿瘤多药耐药机制的研究进展[J].中国肿瘤,2016,25(4):272-277.]
[7]Tang Deng,Zhang Chenli,Ma Linna,et al.Effect of curcumin on migration and invasion of gastric adenocarcinoma cells with different differentiation degrees[J].Journal of Clinical and Experimental Pathology,2018,34(04):424-429.[唐邓,张晨丽,马琳娜,等.姜黄素对不同分化程度胃腺癌细胞迁移侵袭能力的影响[J].临床与实验病理学杂志,2018,34(04):424-429.]
[8]Zhao Yongqiang,Tian Dezeng,Wei Xiaohua,et al.Study of curcumin on radiosensitivity of human papillomatous thyroid carcinoma cell line TP C-1[J].Chinese Journal of Radiation Oncology,2016,25(8):886-890.[赵永强,田德增,魏晓华,等.姜黄素增强人乳头瘤状甲状腺癌细胞TP C-1放射敏感性的研究[J].中华放射肿瘤学杂志,2016,25(8):886-890.]
[9]Han Wei,Lei Yongsheng.Progress in the study of the mechanism of curcumin in the treatment of urogenital cancer[J].Modern Medicine and Clinic,2016,31(2):260-264.[韩伟,雷勇胜.姜黄素治疗泌尿生殖系统癌症的作用机制研究进展[J].现代药物与临床,2016,31(2):260-264.]
[10]Li Xingjia,Xu Yijiao,Chen Guofang,et al.Inhibitory effect of curcumin on proliferation of adriamycin-resistant thyroid cancer cell line HTh74Rdox and its mechanism[J].Jiangsu Medicine,2016,42(24):2653-2656.[李兴佳,徐一娇,陈国芳,等.姜黄素对甲状腺未分化癌阿霉素耐药细胞株HTh74Rdox的增殖抑制作用及其机制[J].江苏医药,2016,42(24):2653-2656.]
[11]Yan Meidi,Cen Xueying.Effect of curcumin on proliferation and apoptosis of thyroid cancer cell line SW579[J].Chinese General Medicine,2015,13(3):396-398.[严梅娣,岑雪英.姜黄素对甲状腺癌细胞SW579增殖和凋亡的影响[J].中华全科医学,2015,13(3):396-398.]
[12]Li Jianjun,Yu Tingting,Shan Li.Advances in the treatment of non-small cell lung cancer with multi-target drugs[J].Medical Review,2017,23(2):275-280.[李建军,俞婷婷,单莉.多靶点药物治疗非小细胞肺癌的研究进展[J].医学综述,2017,23(2):275-280.]
[13]Wu Meng,Xie Yongli,Cen Shan,et al.Advances in multitarget drug therapy for prostate cancer[J].Chinese Medical Biotechnology,2016,11(06):526-531.[吴萌,谢永丽,岑山,等.多靶点药物治疗前列腺癌研究进展[J].中国医药生物技术,2016,11(06):526-531.]
[14]Zhang YY,Zhang J,Zhang WJ,et al.Expression and significance of TFF3 and ERK pathway in papillary thyroid carcinoma[J].Modern Preventive Medicine,2016,43(5):875-879.[张芸芸,张静,张文静,等.TFF3和ERK通路在甲状腺乳头状癌中的表达和意义[J].现代预防医学,2016,43(5):875-879.]
[15]Niko Hensel,Svetlana Baskal,Lisa Marie Walter,et al.ERK and ROCK functionally interact in a signaling network that is compensationally upregulated in spinal muscular atrophy[J].Neurobiology of Disease,2017,22(6):143-147.
[16]Mao Jiading,Hu Di,Wu Pei.The role of gastrin in regulating ERKsignaling pathway in promoting the proliferation of CACO2 cells in colorectal cancer[J].Chinese Clinical Pharmacology and Therapeutics,2011,22(04):401-405.[茆家定,胡迪,吴佩.胃泌素调控ERK信号通路在促进大肠癌CACO2细胞增殖中的作用[J].中国临床药理学与治疗学,2017,22(04):401-405.]
[17]Li Peng,Gong Xingji,Cao Wen.Apoptosis of undifferentiated thyroid carcinoma FRO cells involved in ERK pathway and regulation of arsenic trioxide[J].Cancer Progress,2016,14(9):879-881.[李鹏,宫兴基,曹文.ERK通路参与的未分化甲状腺癌FRO细胞凋亡过程及三氧化二砷的调控作用研究[J].癌症进展,2016,14(9):879-881.]
[18]Zhang Xinyu,Chen Peng,Tao Xuanchen.Progress in the study of the interaction between arsenic trioxide and histone modification[J].Journal of Gastroenterology and Hepatology,2018,27(03):351-355.
[19]Yuko E,Katsuhiko Y,Yuichi K,et al.Green tea extract attenuates MNU-induced photoreceptor cell apoptosis via suppression of heme oxygenase-1[J].Journal of Toxicologic Pathology,2016,29(1):61-65.