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医药导报  2019, Vol. 38 Issue (6): 693-700    DOI: 10.3870/j.issn.1004-0781.2019.06.002
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消癌平注射液调节核受体增强紫杉醇抑制卵巢癌A2780细胞增殖的机制*
张湘奇1(),陈君君2,杨姣1,2,石美智2,祁美娟3,肖霄2,王洪斌4,韩永龙1,2()
1.上海交通大学附属第六人民医院药剂科,上海 200233
2.上海健康医学院附属第六人民医院东院药剂科,上海 201306
3.上海海洋大学食品学院,上海 201306
4.北加州大学药学院,美国加利福尼亚 CA95757
Enhances of the Inhibitory Effect of Paclitaxel on the Proliferation of Ovarian Cancer A2780 Cell by Xiaoaiping Injection via Modulating Nuclear Receptors
Xiangqi ZHANG1(),Junjun CHEN2,Jiao YANG1,2,Meizhi SHI2,Meijuan QI3,Xiao XIAO2,Hongbin WANG4,Yonglong HAN1,2()
1.Department of Pharmacy, Shanghai Jiaotong University Affiliated the Sixth People's Hospital, Shanghai 200233,China
2.Department of Pharmacy, Shanghai University of Medicine & Health Sciences Affiliated the Sixth People's Hospital East Campus, Shanghai 201306,China
3.College of Food Sciences & Technology, Shanghai Ocean University, Shanghai 201306,China
4.College of Pharmacy, California North State University, California CA95757, USA
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摘要 

目的 探讨消癌平注射液通过调节核受体PXR/CAR增强紫杉醇抑制卵巢癌A2780细胞增殖的作用机制。方法 分别将20,40,80,160 mg·mL-1消癌平注射液,10,40,80,160 nmol·L-1紫杉醇单用,或两药联用于人卵巢癌细胞A2780细胞株,采用噻唑蓝(MTT)法检测药物对细胞的抑制率,倒置相差显微镜观察药物对细胞形态的影响,逆转录-聚合酶链反应(RT-PCR)法检测药物对PXR、CAR及下游药物代谢酶CYP2C8、CYP3A4、CYP3A5和转运体P-gp的mRNA表达的影响,Western blotting法检测药物对PXR、CYP2C8蛋白表达的影响。结果 消癌平注射液处理卵巢癌A2780细胞后,细胞增殖受到抑制,且随药物浓度增加、作用时间延长,抑制率显著增加;与紫杉醇单用组比较,消癌平注射液联用紫杉醇后对细胞增殖的抑制作用更为显著;RT-PCR和Western blotting结果显示消癌平注射液可抑制紫杉醇激活的PXR及下游代谢酶CYP2C8的mRNA和蛋白的表达。结论 消癌平注射液与紫杉醇联用能够增强紫杉醇对卵巢癌A2780细胞增殖的抑制作用,其机制可能与消癌平注射液抑制与药物代谢相关的核受体PXR和下游药物代谢酶CYP2C8表达有关。

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张湘奇
陈君君
杨姣
石美智
祁美娟
肖霄
王洪斌
韩永龙
关键词 消癌平注射液紫杉醇孕烷X受体药物代谢CYP2C8酶    
Abstract

Objective To explore the mechanism of Xiaoaiping injection enhancing the inhibitory effect of paclitaxel on the proliferation of ovarian cancer cell A2780. Methods Ovarian cancer cell line A2780 were treated by Xiaoaiping injection (20,40,80,160 mg·mL-1) or paclitaxel(10,40,80,160 nmol·L-1) or Xiaoaiping injection combined with paclitaxel, respectively. The inhibition ratios of different drugs on the proliferation of A2780 cells were measured by MTT method. The changes in cell morphology after drug treatment were monitored by microscope. The mRNA expression of PXR, CAR and CYP2C8, CYP3A4, CYP3A5, transporter P-gp were detected by RT-PCR. The protein expression of PXR, CYP2C8 were measured by Western blotting. Results Xiaoaiping injection inhibits the proliferation of A2780 cells in a time- and concentration-dependent manner. Compared with paclitaxel treatment alone, Xiaoaiping injection combined with paclitaxel exhibit more significant inhibitory effect on A2780 cell proliferation RT-PCR and Western blotting results revealed that Xiaoaiping injection can significantly inhibit paclitaxel-induced over expression of PXR and CYP2C8. Conclusion Xiaoaiping injection can significantly enhance the inhibitory effect of paclitaxel on the proliferation of ovarian cancer A2780 cells. The mechanism may be associated with the inhibitory effect of Xiaoaiping on paclitaxel-induced overexpression of nuclear receptor PXR and its downstream drug metabolic enzyme CYP2C8.

Key wordsXiaoaiping injection    Paclitaxel    PXR    Drug metabolism    Enzyme    CYP2C8
收稿日期: 2018-09-12      出版日期: 2019-06-11
基金资助:*国家自然科学基金资助项目(81773949);上海市卫生和计划生育委员会科研课题青年项目(20174Y0023);上海健康医学院种子基金项目(HMSF-17-22-018,HMSF-17-22-020);浦东新区卫生计生科技项目(PW2017E-2);浦东新区民生科研专项资金(PKJ2017-Y07)
引用本文:   
张湘奇,陈君君,杨姣,石美智,祁美娟,肖霄,王洪斌,韩永龙. 消癌平注射液调节核受体增强紫杉醇抑制卵巢癌A2780细胞增殖的机制*[J]. 医药导报, 2019, 38(6): 693-700.
Xiangqi ZHANG,Junjun CHEN,Jiao YANG,Meizhi SHI,Meijuan QI,Xiao XIAO,Hongbin WANG,Yonglong HAN. Enhances of the Inhibitory Effect of Paclitaxel on the Proliferation of Ovarian Cancer A2780 Cell by Xiaoaiping Injection via Modulating Nuclear Receptors. Herald of Medicine, 2019, 38(6): 693-700.
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http://www.yydbzz.com/CN/10.3870/j.issn.1004-0781.2019.06.002      或      http://www.yydbzz.com/CN/Y2019/V38/I6/693
基因名称 引物序列(5'→3')
PXR Forward :TGTGAGTGCGTGTGTGTGAT
Reverse :TGATTGTCAGCGTAGCCTTG
CAR Forward :GCTGCAAGAGGAGATGGCACTG
Reverse :CAGCGGCATCATGGCAGACAG
CYP2C8 Forward :TCCTGTGTTCACCGTGTATT
Reverse :TCAATCAGGGCTTCCTTCAC
CYP3A4 Forward :TCATTGCTGTCTCCAACCTTCACC
Reverse :GGCTTGCCTGTCTCTGCTTCC
CYP3A5 Forward :CCGACGTGATCAGAACAGTGCTAG
Reverse :TGGTGAAGGTTGGAGACAGCAATG
P-gp Forward :GATTGCTCACCGCCTGTCCAC
Reverse :CGTGCCATGCTCCTTGACTCTG
表1  RT-PCR扩增所用的基因引物序列
图1  XAP对A2780细胞增殖的抑制作用($\bar{x}±s$,n=3)
与24 h比较,*1P<0.05,*2P<0.01
图2  PTX及与XAP联用24和48 h对A2780细胞增殖的抑制作用($\bar{x}±s$,n=3)
与PTX组比较,*1P<0.05,*2P<0.01
图3  XAP和PTX作用于A2780细胞24和48 h后形态(×200)
图4  XAP和PTX作用于A2780细胞后PXR的mRNA水平($\bar{x}±s$,n=3)
与对照组比较,*1P<0.01;与PTX组比较,*2P<0.01
图5  XAP和PTX作用于A2780细胞后CAR的mRNA水平($\bar{x}±s$,n=3)
图6  XAP和PTX作用于A2780细胞后CYP2C8的mRNA水平($\bar{x}±s$,n=3)
与对照组比较,*1P<0.01;与PTX组比较,*2P<0.01
图7  XAP和PTX作用于A2780细胞后CYP3A4的mRNA水平($\bar{x}±s$,n=3)
与对照组比较,*1P<0.05
图8  XAP和PTX作用于A2780细胞后CYP3A5 mRNA水平($\bar{x}±s$,n=3)
图9  XAP和PTX作用于A2780细胞后P-gp mRNA水平($\bar{x}±s$,n=3)
图10  XAP和PTX作用于A2780细胞24 h后PXR、CYP2C8蛋白表达水平($\bar{x}±s$,n=3)
图11  XAP和PTX作用于A2780细胞48 h后PXR、CYP2C8蛋白表达水平($\bar{x}±s$,n=3)
与对照组比较,*1P<0.05;与PTX组比较,*2P<0.05
[1] ZHU R J,SHEN X L,DAI L L,et al.Total aglycones from Marsdenia tenacissima increases antitumor efficacy of paclitaxel in nude mice[J].Molecules (Basel,Switzerland),2014,19(9):13965-13975.
doi: 10.3390/molecules190913965
[2] HAN S Y,DING H R,ZHAO W,et al.Enhancement of gefitinib-induced growth inhibition by Marsdenia tenacissima extract in non-small cell lung cancer cells expressing wild or mutant EGFR[J].BMC Complement Altern Med,2014,14:165.
doi: 10.1186/1472-6882-14-165
[3] HUANG T,GONG W H,ZOU C P,et al.Marsdenia tenaci-ssima extract sensitizes MG63 cells to doxorubicin-induced apoptosis[J].Genet Mol Res,2014,13(1):354-362.
doi: 10.4238/2014.January.21.3
[4] GARCIA-MARTIN E,PIZARRO R M,MARTINEZ C,et al.Acquired resistance to the anticancer drug paclitaxel is associated with induction of cytochrome P450 2C8[J].Pharmacogenomics,2006,7(4):575-85.
doi: 10.2217/14622416.7.4.575 pmid: 3222216753005
[5] HARMSEN S,MEIJERMAN I,BEIJNEN J H,et al.Nuclear receptor mediated induction of cytochrome P450 3A4 by anticancer drugs:a key role for the pregnane X receptor[J].Cancer Chemother Pharmacol,2009,64(1):35-43.
doi: 10.1007/s00280-008-0842-3
[6] KUDO T,OZAKI Y,KUSANO T,et al.Effect of buffer conditions on CYP2C8-mediated paclitaxel 6alpha-hydroxylation and CYP3A4-mediated triazolam alpha- and 4-hydroxylation by human liver microsomes[J].Xenobiotica,2016,46(3):241-246.
doi: 10.3109/00498254.2015.1071502
[7] ROCHAT B.Role of cytochrome P450 activity in the fate of anticancer agents and in drug resistance:focus on tamoxifen,paclitaxel and imatinib metabolism[J].Clin Pharmacokinet,2005,44(4):349-366.
doi: 10.2165/00003088-200544040-00002
[8] CHAI X,ZENG S,XIE W.Nuclear receptors PXR and CAR:implications for drug metabolism regulation,pharmacogenomics and beyond[J].Expert Opin Drug Metabol Toxicol,2013,9(3):253-266.
doi: 10.1517/17425255.2013.754010
[9] CHEN Y,TANG Y,GUO C,et al.Nuclear receptors in the multidrug resistance through the regulation of drug-metabolizing enzymes and drug transporters[J].Biochem Pharmacol,2012,83(8):1112-1126.
doi: 10.1016/j.bcp.2012.01.030
[10] KOUTSOUNAS I,PATSOURIS E,THEOCHARIS S.Pre-gnane X receptor and human malignancy[J].Histol Histopathol,2013,28(4):405-420.
[11] MASUYAMA H,NAKAMURA K,NOBUMOTO E,et al.Inhibition of pregnane X receptor pathway contributes to the cell growth inhibition and apoptosis of anticancer agents in ovarian cancer cells[J].Int J Oncol,2016,49(3):1211-1220.
doi: 10.3892/ijo.2016.3611
[12] 吴畏. 1例卵巢癌化疗患者神经毒性治疗的药学监护[J].医药导报,2014,33(12):1650-1652.
doi: 10.3870/yydb.2014.12.031
[13] 邓艾平,王奕,刘珏,等.叶酸壳寡糖修饰的聚乳酸-羟基乙酸共聚物纳米粒对耐紫杉醇人卵巢癌细胞增殖的影响[J].医药导报,2018,37(6):662-666.
[14] ELSERAFI M M,ZEENELDIN A A,ABDELSALAM I M,et al.First-line paclitaxel and cisplatin used sequentially or in combination in metastatic breast cancer:a phase II randomized study[J].J Egypt Natl Cancer Inst,2018,30(1):11-20.
[15] 杨蕾,张振海,贾晓斌.人参稀有皂苷组分联合紫杉醇治疗A549肺癌的实验研究[J].中国中药杂志,2018,43(7):1446-1452.
[16] 霍小蕾,韩玲娜,裴振,等.香菇多糖联合紫杉醇对食管癌细胞生物学行为的影响[J].东南大学学报(医学版),2017,36(6):897-900.
[17] 卢晨欣,孙警辉,伍春莲.白藜芦醇与紫杉醇联合用药对人喉癌Hep-2细胞凋亡机制的研究[J].中国中药杂志,2016,41(3):476-83.
[18] 胡赤丁. 消癌平治疗晚期恶性肿瘤33例[J].医药导报,2001,20(10):623.
[19] FAN W,SUN L,ZHOU J Q,et al.Marsdenia tenacissima extract induces G0/G1 cell cycle arrest in human esophageal carcinoma cells by inhibiting mitogen-activated protein kinase (MAPK) signaling pathway[J].Chin J Nat Med,2015,13(6):428-437.
[20] 刘丽雅,韩永龙,余奇,等.消癌平注射液等4种抗肿瘤中药注射剂对人肝微粒体中CYP450酶7种亚型的体外抑制作用研究[J].中国临床药理学与治疗学,2014,19(5):522-527.
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