蟾毒灵体外抑制人舌鳞癌Tca8113细胞增殖和促凋亡作用及对Na+/K+-ATP酶的影响

doi:10.3969/j.issn.1000-8535.2017.02.003

摘要/Abstract

摘要： 目的探索蟾毒灵对舌鳞状细胞癌Tca8113细胞增殖、凋亡的影响及可能作用机制。方法以人舌鳞癌Tca8113细胞为研究对象,MTT法检测10、20、40、80、160 nmol/L浓度蟾毒灵体外抑制Tca8113细胞增殖的活性;检测蟾毒灵干预下肿瘤细胞Na⁺-K⁺-ATP酶活性的变化;Western blot发检测Bcl-2、Bax、caspase-3蛋白表达。结果蟾毒灵有抑制Tca8113细胞的活性,且呈剂量-时间依赖性;在蟾毒灵干预下Tca8113细胞Na⁺-K⁺-ATP酶收到抑制;Western blot结果显示凋亡相关Bax、caspase-3蛋白表达上调,Bcl-2蛋白表达下调。结论蟾毒灵通过抑制细胞膜Na⁺-K⁺-ATP酶活性,通过调节Bcl-2凋亡通路的相关蛋白,最终激活caspase-3,诱导人舌鳞癌Tca8113细胞凋亡。

关键词: 蟾毒灵, 舌鳞状细胞癌, Na⁺-K⁺-ATP酶, 细胞凋亡

Abstract: Objective To explore the effects of bufalin on proliferation and apoptosis of tongue squamous cell carcinoma Tca8113 cells and its possible mechanism. Methods Tca8113 cells were treated with 10, 20, 40, 80 and 160 nmol/L Tca8113 cells in vitro. MTT assay was used to detect the inhibitory effect of bufalin on the proliferation of Tca8113 cells; And the activity of Na⁺-K⁺-ATPase in tumor cells was detected by the interference of bufalin; The expression of Bcl-2, Bax and caspase-3 protein was detected by Western blot. Results Bufalin inhibited the activity of Tca8113 cells in a dose-and time-dependent manner; Na⁺-K⁺-ATPase in Tca8113 cells was inhibited by bufalin; The results of Western blot showed that the expression of Bax and caspase-3 protein was up-regulated and the expression of Bcl-2 protein was down-regulated. Conclusion Bufalin induced the apoptosis of human tongue squamous cell carcinoma Tca8113 cells by inhibiting the activity of Na⁺-K⁺-ATPase and regulating the related proteins of Bcl-2 apoptosis pathway, finally activating caspase-3.

Key words: Bufalin, Tongue squamous cell carcinoma, Na⁺-K⁺-ATPase, Cell apoptosis

杨瑞琨, 熊建辉. 蟾毒灵体外抑制人舌鳞癌Tca8113细胞增殖和促凋亡作用及对Na⁺/K⁺-ATP酶的影响[J]. 广州医药, 2017, 48(2): 10-13.

YANG Ruikun, XIONG Jianhui. Apoptosis of Tca8113 cell induced by bufalin and the inference to Na⁺-K⁺-ATPase[J]. Guangzhou Medical Journal, 2017, 48(2): 10-13.

参考文献

[1] 涂玉绮. 穿心莲内酯对人舌鳞状细胞癌Tca8113细胞株增殖和抑制影响的实验研究[D]. 南昌大学医学院, 2013.
[2] 武云霞. 选择性环氧化酶-2抑制剂影响舌鳞癌血管生成的研究[D]. 山西医科大学, 2007.
[3] ALEVIZOPOULOS K, CALOGEROPOULOU T, LANG F, et al. Na⁺/K⁺ ATPase Inhibitors in cancer[J]. Curr Drug Targets, 2014, 15(10): 988-1000.
[4] 王柏琦. 穿心莲内酯对鼻咽癌细胞增殖、凋亡及Na⁺/K⁺-ATP酶活性的影响[D]. 南华大学, 2014.
[5] DURLACHER C T, CHOW K, CHEN X W, et al. Targeting Na⁺/K⁺-translocating adenosine triphosphatase in cancer treatment[J]. Clin Exp Pharmacol Physiol, 2015, 42(5): 427-443.
[6] 吕银鹏, 周辉, 宋佳欣. 蟾蜍灵联合顺铂对人舌鳞状细胞癌Tca8113细胞增殖及凋亡的影响[J]. 山东医药, 2014 (46): 17-20.
[7] 占跃晨. 洋地黄类药物诱导肿瘤细胞自噬及相关激酶信号通路的研究[D]. 北京协和医学院中国医学科学院北京协和医学院清华大学医学部中国医学科学院, 2014.
[8] 陈果. 强心苷类化合物抗肿瘤作用及机制[D]. 桂林医学院, 2011.
[9] MIJATOVIC T, DUFRASNE F, KISS R. Na⁺/K⁺-ATPase and cancer[J]. Pharm Pat Anal, 2012, 1(1): 91-106.
[10] WANG H Y, O'DOHERTY G A. Modulators of Na⁺/K⁺-ATPase: a patent review[J]. Expert Opin Ther Pat, 2012, 22(6): 587-605.
[11] 张力, 张瀚彬, 陈淼,等. 低氘水抑制宫颈癌Hela细胞增殖和侵袭能力的研究[J]. 中国医药导报, 2014, 11(9): 20-25.
[12] CHAVES A L F, OLIVEIRA De LIMA P, SOARES J M A, et al. Effects of digoxin and Na, K-ATPase immunoexpression on human oral squamous carcinomas[J]. Anticancer Res, 2014, 34(10): 5397-5404.
[13] CHEN D, SONG M, MOHAMAD O, et al. Inhibition of Na⁺/K⁺-ATPase induces hybrid cell death and enhanced sensitivity to chemotherapy in human glioblastoma cells[J]. BMC Cancer, 2014, 14: 716/1-716/15.
[14] GARCIA D G, CAIRE De CASTRO-FARIA-NETO H, Ignacio Da Silva C, et al. Na/K-ATPase as a target for anticancer drugs: studies with perillyl alcohol[J]. Mol Cancer, 2015, 14: 1-14.
[15] LEV M C, KARLISH S J D, GARTY H. Cardiac glycosides induced toxicity in human cells expressing α1-, α2-, or α3-isoforms of Na-K-ATPase[J]. Am J Physiol, 2015, 309(1, Pt. 1): C126-C135.
[16] MILUTINOVIC S, HEYNEN-GENEL S, CHAO E, et al. Cardiac glycosides activate the tumor suppressor and viral restriction factor promyelocytic leukemia protein (PML)[J]. PLoS One, 2016, 11(3): e0152692/1-e0152692/16.
[17] YUE Q, ZHEN H, HUANG M, et al. Proteasome inhibition contributed to the cytotoxicity of arenobufagin after its binding with Na, K-ATPase in human cervical carcinoma HeLa cells[J]. PLoS One, 2016, 11(7): e0159034/1-e0159034/28.
[18] ZEINO M, BRENK R, GRUBER L, et al. Cytotoxicity of cardiotonic steroids in sensitive and multidrug-resistant leukemia cells and the link with Na⁺/K⁺-ATPase[J]. J Steroid Biochem. Mol Biol, 2015, 150: 97-111.