[1]雷越,万婕,李丹丹,等.下调FoxM1通过激活JNK/线粒体通路增加人鼻咽癌细胞对紫杉醇的敏感性[J].第三军医大学学报,2018,40(09):780-787.
 LEI Yue,WAN Jie,LI Dandan,et al.Down-regulation of FoxM1 enhances sensitivity of human nasopharyngeal carcinoma cells to paclitaxel via activation of JNK/mitochondrial pathway[J].J Third Mil Med Univ,2018,40(09):780-787.
点击复制

下调FoxM1通过激活JNK/线粒体通路增加人鼻咽癌细胞对紫杉醇的敏感性(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第09期
页码:
780-787
栏目:
基础医学
出版日期:
2018-05-15

文章信息/Info

Title:
Down-regulation of FoxM1 enhances sensitivity of human nasopharyngeal carcinoma cells to paclitaxel via activation of JNK/mitochondrial pathway
作者:
雷越万婕李丹丹叶琳刘亚男祝琳陈鸿雁
重庆医科大学附属第一医院:耳鼻喉头颈外科,胸心外科;江西省九江市第一人民医院耳鼻喉头颈外科;西南医科大学附属医院耳鼻咽喉头颈外科
Author(s):
LEI Yue WAN Jie LI Dandan YE Lin LIU Yanan ZHU Lin CHEN Hongyan

Department of Otolaryngology, Head and Neck Surgery, Department of Cardiothoracic Surgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016;-Department of Otolaryngology, Head and Neck Surgery, Jiujiang First People’s Hospital, Jiujiang, Jiangxi Province, 332000; Department of Otolaryngology, Head and Neck Surgery, the Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan Province, 646000, China

关键词:
鼻咽癌转录因子叉头框M1凋亡化疗敏感性紫杉醇JNK/线粒体通路
Keywords:
nasopharyngeal carcinoma forkhead box protein M1 apoptosis chemosensitivity paclitaxel JNK/mitochondrial pathway
分类号:
R73-362; R739.63; R979.19
文献标志码:
A
摘要:

目的    探讨特异性小干扰RNA(small interfering RNA, siRNA)沉默转录因子叉头框M1(FoxM1)对人鼻咽癌HONE-1细胞增殖、凋亡、紫杉醇敏感性的影响及可能的分子机制。方法    采用siRNA靶向沉默鼻咽癌HONE-1细胞FoxM1表达并通过RT-PCR、实时定量PCR、Western blot检测转染后FoxM1的表达水平。siRNA转染后细胞的增殖和对紫杉醇的敏感性采用MTT法检测,流式细胞仪检测细胞周期分布,Annexin V-FITC/PI双染法检测细胞凋亡率。Western blot检测Ki67、CyclinE1、多药耐药基因1(multidrug resistance 1,MDR1)及c-Jun氨基末端激酶(c-Jun N-terminal kinase, JNK)、p-JNK、c-Jun、p-c-Jun、凋亡相关蛋白Bax、Bcl-2的表达水平。结果    转染后FoxM1 mRNA及蛋白的表达均被下调(P<0.01)。siRNA转染组细胞增殖速率减缓(P<0.05),Ki67表达降低(P<0.05)。siRNA转染组G1期细胞比例增加(P<0.01),S期比例减低(P<0.05), Cyclin E1低表达(P<0.01)。同时,siRNA+紫杉醇组凋亡率明显高于阴性及空白对照+紫杉醇组(P<0.01)。siRNA转染组细胞MDR1水平下降(P<0.05),对紫杉醇的敏感性较两对照组明显增强(P<0.01)。siRNA+紫杉醇组与阴性对照+紫杉醇组相比pJNK1、p-c-Jun、Bax表达上调(P<0.01),Bcl-2表达下调(P<0.01)。结论    特异性siRNA沉默FoxM1表达可以影响JNK/线粒体通路活性,抑制鼻咽癌HONE-1细胞增殖,促进其凋亡,提高其对紫杉醇的敏感性。

Abstract:

Objective    To determine the effects of down-regulating Forkhead box M1 gene (FoxM1) on the proliferation, apoptosis and sensitivity to paclitaxel in human nasopharyngeal carcinoma cell line HONE1, and investigate the possible molecular mechanism. Methods    A sequence-specific small interfering RNA (siRNA) targeting FoxM1 was used to transfect HONE-1 cells, then the expression of FoxM1 was detected by quantitative reverse transcription-PCR (qRT-PCR), RT-PCR and Western blotting. The proliferation and chemosensitivity to paclitaxel cells were detected by MTT assay. Flow cytometry was adopted to measure the cell cycle distribution of transfected HONE-1 cells. The apoptotic rates were examined by Annexin V-FITC/PI staining assay. And the expression levels of Ki67, Cyclin E1, multidrug resistance 1 (MDR1), c-Jun Nterminal kinase (JNK), p-JNK, c-Jun, p-c-Jun, and apoptosis-related proteins Bax and Bcl-2 were detected by Western blotting. Results    FoxM1-siRNA transfection decreased the expression level of FoxM1 gene in HONE-1 cells obviously (P<0.01). Compared with the control cells, the transfected HONE-1 cells had markedly decreased proliferation rate (P<0.05), much lower Ki67 expression (P<0.05), increased percentage of the cells in G1 phase (P<0.01) while less cells in S phase (P<0.05), and downregulated expression of Cyclin E1 ( P<0.01). Meanwhile the apoptotic rate was significantly higher in the siRNA/paclitaxel cells than negative and blank control/paclitaxel cells (P<0.01). What’s more, the transfection obviously decreased the MDR1 level (P<0.05), and significantly enhanced the sensitivity of HONE-1 cells to paclitaxel (P<0.01), when compared with the control groups. In siRNA/paclitaxel group, the expression levels of p-JNK1, p-c-Jun and Bax were much higher than the negative control/paclitaxel group (all P<0.01), whereas that of Bcl-2 was down-regulated (P<0.01). Conclusion    The siRNA targeting FoxM1 in HONE-1 cells could effectively regulate the activity of JNK/mitochondrial pathway, inhibit the proliferation, promote the apoptosis, and ultimately enhance the sensitivity to paclitaxel.

参考文献/References:

[1]SIEGEL R L, MILLER K D, JEMAL A. Cancer statistics, 2017[J]. CA Cancer J Clin, 2017, 67(1):7-30.DOI:10.3322/caac.21387.
[2]CHEN W, ZHENG R, BAADE P D, et al.Cancer statistics in China, 2015[J]. CA Cancer J Clin,2016,66(2):115-132.DOI:10.3322/caac.21338.
[3]CHEN C, WANG F H, AN X, et al.Triplet combination with paclitaxel, cisplatin and 5FU is effective in metastatic and/or recurrent nasopharyngeal carcinoma[J]. Cancer Chemoth Pharm, 2012, 71(2):371-378.DOI:10.1007/s00280-012-2020-x.
[4]HE X Y, HU C S, YING H M, et al.Paclitaxel with cisplatin in concurrent chemoradiotherapy for locally advanced nasopharyngeal carcinoma[J]. Eur Arch Otorhinolaryngol, 2010, 267(5): 773-778.DOI:10.1007/s00405-009-1112-7.
[5]SU S F, HAN F, ZHAO C, et al. Long-term outcomes of earlystage nasopharyngeal carcinoma patients treated with intensity-modulated radiotherapy alone[J]. Int J Radiat Oncol Biol Phys, 2012, 82(1):327-333.DOI:10.1016/j.ijrobp.2010.09.011.
[6]KAMRAN S C, RIAZ N, LEE N. Nasopharyngeal carcinoma[J]. Surg Oncol Clin N Am, 2015, 24(3):547-561.DOI:10.1016/j.soc.2015.03.008.
[7]FAYETTE J, MONTELLA A, CHABAUD S, et al. Paclitaxel is effective in relapsed head and neck squamous cell carcinoma: A retrospective study of 66 patients at a single institution[J]. Anticancer drug, 2010, 21(5):553-558.DOI:10.1097/CAD.0b013e3283388e60.
[8]KWOK J M, PECK B, MONTEIRO L J, et al. FOXM1 confers acquired cisplatin resistancein breastcancer cells[J]. Mol Cancer Res, 2010, 8(1):24-34.DOI:10.1158/1541-7786.MCR-09-0432.
[9]WANG Y, WEN L, ZHAO S H, et al. FoxM1 expression is significantly associated withcisplatinbased chemotherapy resistance and poor prognosis in advanced non-small celllung cancerpatients[J]. Lung Cancer, 2013, 79(2):173-179.DOI:10.1016/j.lungcan.2012.10.019.
[10]OKADA K, FUJIWARA Y, TAKAHASHI T, et al. Overexpression of forkhead box M1 transcriptionfactor (FOXM1) is a potential prognostic marker and enhances chemoresistance for docetaxel ingastric cancer[J]. Ann Surg Oncol,2013,20(3):1035-1043.DOI:10.1245/s10434-012-2680-0.
[11]WANG K, ZHU X, ZHANG K, et al. FoxM1 inhibition enhances chemosensitivity of docetaxelresistant A549 cells to docetaxel via activation of JNK/mitochondrial pathway[J]. Acta Biochim Biophys Sin (Shanghai), 2016, 48(9): 804-809.DOI:10.1093/abbs/gmw072.
[12]ZHAO F, SIU M K, JIANG L, et al. Overexpression of forkheadbox protein M1 (FOXM1) in ovarian cancer correlates with poor patient survival and contributes to paclitaxel resistance[J]. PLoS ONE, 2014, 9(11):e113478.DOI:10.1371/journal.pone.0113478.
[13]JIANG L, WANG P, CHEN H. Overexpression of FOXM1 is associated with metastases of nasopharyngeal carcinoma[J]. Ups J Med Sci, 2014, 119(4):324-332.DOI:10.3109/03009734.2014.960053.
[14]魏蕾,江黎珠, 于超,等. 下调FoxM1表达对人喉癌Hep-2细胞顺铂敏感性的影响[J]. 第三军医大学学报, 2015, 37(16):1603-1608.DOI:10.16016/j.10005404.201411182.
WEI L, JIANG L Z, YU C, et al. FoxM1 down-regulation promotes sensitivity of laryngeal carcinoma Hep2 cells to cisplatin[J]. J Third Mil Med Univ, 2015, 37(16): 1603-1608.DOI:10.16016/j.10005404.201411182.
[15]JIANG L, WANG P, CHEN L, et al. Downregulation of FoxM1 by thiostrepton or small interfering RNA inhibits proliferation, transformation ability and angiogenesis, and induces apoptosis of nasopharyngeal carcinoma cells[J]. Int J Clin Exp Pathol, 2014, 7(9):5450-5460.
[16]权芳, 张少强, 白艳霞,等. 白藜芦醇对缺氧环境中鼻咽癌细胞株CNE2的化疗增敏作用[J]. 中西医结合学报,2009,7(10):952-957.
QUAN F, ZHANG S Q, BAI Y X, et al. Resveratrol increases sensitivity of CNE2 cells to chemotherapeutic drugs under hypoxia[J]. J Chin Integr Med, 2009,7(10):952-957.
[17]KAESTNER K H, KNOCHEL W, MARTINEZ D E. Unified nomenclature for the wingedhelix/forkhead transcription factors[J]. Genes Dev, 2000, 14(2):142-146.
[18]WU Q F, LIU C, TAI M H, et al. Knockdown of FoxM1 by siRNA interference decreasescell proliferation, induces cell cycle arrest and inhibits cell invasion in MHCC-97H cells in vitro[J]. Acta Pharmacol Sin, 2010, 31(3):361-366.DOI: 10.1038/aps.2010.4.
[19]YU J, WANG X, LI Y, et al.TanshinoneⅡA suppresses gastric cancer cell proliferation and migration by downregulation of FOXM1[J]. Oncol Rep, 2017,37(3):1394-1400.DOI: 10.3892/OR.2017.5408.
[20]PANAYOTOPOULOU E G, M-LLER A K, B-RRIES M, et al.Targeting of apoptotic pathwaysby SMAC or BH3 mimetics distinctly sensitizes paclitaxelresistant triple negative breast cancer cells[J]. Oncotarget, 2017, 8(28):45088-45104.DOI: 10.18632/ONCOTARGET.15125.
[21]JIN H, PARK M H, KIM S M. 3,3′Diindolylmethane potentiates paclitaxelinduced antitumor effects on gastric cancer cells through the Akt/FOXM1 signaling cascade[J].Oncol Rep, 2015, 33(4):2031-2036.DOI:10.3892/or.2015.3758.
[22]HUANG X, QIN J, LU S. Upregulation of miR877 induced by paclitaxel inhibitshepatocellular carcinoma cell proliferation though targeting FOXM1[J]. Int J Clin Exp Pathol, 2015, 8(2):1515-1524.
[23]FREDERIKS C N, LAM S W, GUCHELAAR H J, et al. Genetic polymorphisms andpaclitaxelor docetaxelinduced toxicities: A systematic review[J].Cancer Treat Rev, 2015, 41(10):935-950.DOI:10.1016/j.ctrv.2015.10.010.
[24]ZHAO Y, MU X, DU G. Microtubule-stabilizing agents: New drug discovery and cancer therapy[J]. Pharmacol Ther, 2016, 162:134-143.DOI:10.1016/j.pharmthera.2015.12.006.
[25]HOU Y, ZHU Q, LI Z, et al. The FOXM1-ABCC5 axis contributes to paclitaxel resistance in nasopharyngeal carcinoma cells[J]. Cell Death Dis, 2017, 8(3):e2659.DOI:10.1038/cddis.2017.53.
[26]WANG I C, CHEN Y J, HUGHES D E, et al. FoxM1 regulates transcription of JNK1 to promote the G1/S transition and tumor cell invasiveness[J]. J Biol Chem, 2008, 283(30):20770-20778.DOI:10.1074/jbc.M709892200.
[27]WADA T, PENNINGER J M. Mitogen-activated protein kinases in apoptosisregulation[J]. Oncogene, 2004, 23(16):2838-2849.DOI: 10.1038/sj.onc.1207556.
[28]RAGGATT L J, EVDOKIOU A, FINDLAY D M. Sustained activation of Erk1/2 MAPK and cell growth suppression by the insert-negative, but not the insert-positive isoform of the human calcitonin receptor[J]. J Endocrinol, 2000, 167(1):93-105.
[29]CHATTOPADHYAY S, MACHADO-PINILLA R, MANGUAN-GARC-A C, et al. MKP1/CL100 controls tumor growth and sensitivity to cisplatin in nonsmallcell lung cancer[J]. Oncogene, 2006, 25(23):3335-3345.DOI: 10.1038/sj.onc.1209364.

更新日期/Last Update: 2018-05-11