[1]江雪均,杨建波,曾梅,等.新型吴茱萸碱衍生物抗小细胞肺癌活性效应的初步评价[J].第三军医大学学报,2019,41(10):961-968.
 JIANG Xuejun,YANG Jianbo,ZENG Mei,et al.Antitumor activity of 2 new evodiamine derivatives against small-cell lung cancer cells in vitro[J].J Third Mil Med Univ,2019,41(10):961-968.
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新型吴茱萸碱衍生物抗小细胞肺癌活性效应的初步评价(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第10期
页码:
961-968
栏目:
基础医学
出版日期:
2019-05-30

文章信息/Info

Title:
Antitumor activity of 2 new evodiamine derivatives against small-cell lung cancer cells in vitro
作者:
江雪均杨建波曾梅张景勍胡雪原刘颖菊
重庆医科大学药学院:重庆市生物化学与分子药理学重点实验室,重庆市高校药物工程中心
Author(s):
JIANG Xuejun YANG Jianbo ZENG Mei ZHANG Jingqing HU Xueyuan LIU Yingju

Chongqing Key Laboratory of Biochemistry and Molecular Pharmacology, Chongqing Collegiate Pharmaceutical Engineering Research Center, College of Pharmacy, Chongqing Medical University, Chongqing, 400016, China
 

关键词:
吴茱萸碱硝基衍生物氨基衍生物小细胞肺癌凋亡
Keywords:
evodiamine nitro evodiamine derivatives amino evodiamine derivatives small-cell lung cancer apoptosis  
分类号:
R730.53;R734.2;R931.6
文献标志码:
A
摘要:

目的 研究吴茱萸碱(evodiamine, EVO)的两种衍生物:硝基吴茱萸碱衍生物(10-nitro evodiamine derivatives, END)与氨基吴茱萸碱衍生物(10-amino evodiamine derivatives, EAD)对小细胞肺癌细胞株H1688的抗癌活性及机制。方法 H1688细胞分别用不同浓度的吴茱萸碱EVO、END、EAD处理24、48、72 h后用MTT法检测其存活率;10 μmol/L的EVO、END、EAD处理H1688细胞24 h后,流式细胞术检测其凋亡与周期;同样浓度处理48 h后提取总蛋白,Western blot检测Caspase 12、Caspase 9、Caspase 3、细胞色素C(cytochome C, Cyt C)的表达。结果 MTT结果显示,0.625μmol/L的END或EAD处理24 h后,即可观察到药物对H1688的抑制作用,20 μmol/L EVO、END和EAD处理24、48、72 h后,与0 μmol/L组存活率相比显著降低。END和EAD处理组H1688存活率分别为16.45%和61.20%(PEND=0.000, PEAD=0.046, 24 h)、9.10%和43.37%(PEND=0.000, PEAD=0.000, 48 h)、5.24%和29.55%(PEND=0.000, PEAD=0.000, 72 h);而EVO处理组存活率为60.81%(P=0.038,24 h)、21.99%(P=0.000,48 h)、18.51%(P=0.000,72 h),其中20 μmol/L END对H1688抑制效果最好。10 μmol/L的END、EAD处理H1688 24 h后,细胞凋亡率明显升高,凋亡率分别为54.98%(P=0.000)和19.73%(P=0.029),且分别有79.78%(P=0.001)和77.72%(P=0.001)的细胞被阻滞在G2/M期。END、EAD处理48 h后,与Control组相比,H1688细胞中Caspase 9(PEND=0.011,PEAD=0.026)、Caspase 12(PEND=0.031,PEAD=0.018)、Caspase 3(PEND=0.002,PEAD=0.043)、Cyt C(PEND=0.023,PEAD=0.028)等凋亡相关蛋白均上调。与EVO相比,END对Caspase 3(P=0.026)和Cyt C(P=0.002)作用都明显强于EVO,而EAD对Caspase家族与Cyt C的作用则与EVO类似。结论 EVO的两种衍生物END、EAD都对小细胞肺癌细胞株H1688有体外抗肿瘤作用,这种作用呈时间、浓度依赖性,END体外抗肿瘤效果更好。
 

Abstract:

Objective To investigate the antitumor activity and the underlying mechanisms of 2 new derivatives of evodiamine (EVO), namely nitro evodiamine derivatives (END) and amino evodiamine derivatives (EAD), in cultured small-cell lung cancer (SCLC) cells. Methods Human SCLC cell line H1688 was treated with EVO, END, or EAD at different concentrations for 24, 48, or 72 h, and the changes in the cell viability were assessed using MTT assay. The effects of treatment with 10 μmol/L EVO, END or EAD for 24 h on cell apoptosis and cell cycle were investigated using flow cytometry. Western blotting was performed to detect the changes in the expressions of Caspase-9, Caspase-12, Caspase-3, and cytochrome C (Cyt C) in the cells following treatment with 10 μmol/L EVO, END or EAD for 48 h. Results MTT assay showed that treatment with END or EAD for 24 h at the concentration as low as 0.625 μmol/L was sufficient to reduce the viability of H1688 cells. Treatment of the cells with 20 μmol/L END or EAD significantly lowered the cell viability to 16.45% (P=0.000) and 61.20% (P=0.046) at 24 h, to 9.10% (P=0.000) and 43.37% (P=0.000) at 48 h, and to 5.24% (P=0.000) and 29.55% (P=0.000) at 72 h, respectively; the  viability of EVO-treated cells was 60.81% (P=0.038) at 24 h, 21.99% (P=0.000) at 48 h, and 18.51% (P=0.000) at 72 h. Treatments of the cells with 10 μmol/L END and EAD for 24 h both induced obvious cell apoptosis with apoptotic rates of 54.98% (P=0.000) and 19.73% (P=0.029), respectively, and caused cell cycle arrest in G2/M phase in 77.78% (P=0.001) and 77.72% (P=0.001) of the cells, respectively. Treatments with 10 μmol/L END and EAD for 48 h both significantly upregulated the expression of Caspase-9(P=0.011, 0.026), Caspase-12(P=0.031, 0.018), Caspase-3(P=0.002, 0.043) and Cyt C(P=0.023, 0.028) in H1688 cells. END produced stronger effects than EVO in up-regulating caspase-3(P=0.026) and Cyt C(P=0.002); the effects of EAD on caspase family and Cyt C were similar to those of EVO. Conclusion Both of the two derivatives of EVO, END and EAD, and particularly the former, have concentration- and time-dependent antitumor effects against H1688 cells in vitro. Both END and EAD can cause cell cycle arrest in G2/M phase and induce apoptosis of H1688 cells via the mitochondrial pathway.

参考文献/References:

[1]YANG F, SHI L, LIANG T, et al. Anti-tumor effect of evodiamine by inducing Akt-mediated apoptosis in hepatocellular carcinoma[J]. Biochem Biophys Res Commun, 2017, 485(1): 54-61. DOI:  10.1016/j.bbrc.2017.02.017.
[2]WU W S, CHIEN CC, LIU K H, et al. Evodiamine prevents glioma growth, induces glioblastoma cell apoptosis and cell cycle arrest through JNK activation[J]. Am J Chin Med, 2017, 45(4): 879-899. DOI: 10.1142/S0192415X17500471.
[3]ZHONG Z F, TAN W, WANG S P,et al. Anti-proliferative activity and cell cycle arrest induced by evodiamine on paclitaxel-sensitive and -resistant human ovarian cancer cells[J]. Sci Rep, 2015, 5:  16415. DOI: 10.1038/srep16415.
[4]FANG C S, ZHANG J Q, QI D,et al. Evodiamine induces G2/M arrest and apoptosis via mitochondrial and endoplasmic reticulum pathways in H446 and H1688 human small-cell lung cancer cells[J]. PLoS ONE, 2014, 9(12):  e115204. DOI: 10.1371/journal.pone.0115204.
[5]祁迪, 谭群友, 王如文, 等. 吴茱萸碱抑制PI3K/AKT通路诱导小细胞肺癌H1688和H446细胞凋亡 [J]. 第三军医大学学报, 2016, 38(4): 330-337. DOI: 10.16016/j.1000-5404.201508031.
QI D, TAN Q Y, WANG R W, et al. Evodiamine induces apoptosis of small-cell lung cancer H1688 and H446 cells via inhibiting PI3K/AKT pathway[J]. J Third Mil Med Univ, 2016, 38(4):  330-337. DOI: 10.16016/j.1000-5404.201508031.
[6]徐俊杰, 杨然, 杨芳景, 等. 吴茱萸碱抗肿瘤机制的研究进展[J].上海交通大学学报(医学版), 2018, 38(5): 578-583. DOI: 10.3969/j.issn.1674-8115.2018.05.020.
XU J J, YANG R, YANG F J, et al. Recent advances in the antitumor effects of evodiamine [J]. J Shanghai Jiaotong Univ (Med Sci), 2018, 38(5) : 578-583. DOI:  10.3969/j.issn.1674-8115.2018.05.020
[7]李娜.吴茱萸碱衍生物的合成及其药效学和药代动力学初步评价[D].重庆: 重庆医科大学, 2016.
LI N. The synthesis of evodiamine derivatives and preliminary evaluation of the pharmacodynamics and pharmacokinetics[D].Chongqing:  Chongqing Medical University, 2016.
[8]万坤. 吴茱萸碱衍生物EVB的合成及其纳米粒的初步评价[D]. 重庆: 重庆医科大学, 2014.
WAN K. The synthesis of evodiamine derivative and preliminary evaluation of EVB loaded with nanoparticles[D]. Chongqing:  Chongqing Medical University, 2014.
[9]胡雪原, 李娜, 张景勍, 等. 一种吴茱萸碱衍生物的制备方法:  CN201510725965.1[P]. 2015-12-23.
 HU X Y, LI N, ZHANG J Q, et al.The method for preparing a derivative of evodia officinalis: CN201510725965.1[P].2015-12-23.
[10]姚迪,孙晶晶,刘雷, 等. LC-MS/MS法测定大鼠血浆中的吴茱萸碱、吴茱萸次碱和吴茱萸内酯及其生物利用度的研究[J]. 华西药学杂志, 2014, 29(3): 298-302. DOI: 10.13375/j.cnki.wcjps.2014.03.022.
YAO D, SUN J J, LIU L, et al. Simultaneous determination and absolute oral bioavailability of evodiamine,rutaecarpine and evodin concentration in the rat plasma by LC-MS/MS[J]. West China J Pharm Sci, 2014, 29(3): 298-302. DOI: 10.13375/j.cnki.wcjps.2014.03.022.
[11]晏声蕾, 胡江波, 王薛, 等. 吴茱萸碱油包水型复合纳米乳的药代动力学和在体肠吸收[J]. 第二军医大学学报, 2017, 38(2): 249-252. DOI: 10.16781/j.0258-879x.2017.02.0249.
YAN S L, HU J B, WANG X, et al. Pharmacokinetics and in situintestinal absorption of evodiamine complex water-in-oil nanoemulation[J]. Acad J Second Military Med Univ, 2017, 38(2): 249-252. DOI: 10.16781/j.0258-879x.2017.02.0249.
[12]CHRISTODOULOU M S, SACCHETTI A, RONCHETTI V,et al. Quinazolinecarboline alkaloid evodiamine as scaffold for targeting topoisomerase I and sirtuins[J]. Bioorg Med Chem, 2013, 21(22):  6920-6928. DOI: 10.1016/j.bmc.2013.09.030.
[13]ZHAO N, TIAN K T, CHENG K G,et al. Antiproliferative activity and apoptosis inducing effects of nitric oxide donating derivatives of evodiamine[J]. Bioorg Med Chem, 2016, 24(13):  2971-2978. DOI: 10.1016/j.bmc.2016.05.001.
[14]SONG S, CHEN Z, LI S, et al. Design, synthesis and evaluation of N13-substituted evodiamine derivatives against human cancer cell lines[J]. Molecules, 2013, 18(12):  15750-15768. DOI: 10.3390/molecules181215750.
[15]CHEN T C, CHIEN CC, WU M S, et al. Evodiamine from Evodia rutaecarpa induces apoptosis via activation of JNK and PERK in human ovarian cancer cells[J]. Phytomedicine, 2016, 23(1):  68-78. DOI: 10.1016/j.phymed.2015.12.003.
[16]WEI W, CHEN H, WANG Z, et al. Enhanced antitumor efficacy of gemcitabine by evodiamine on pancreatic cancer via regulating PI3K/Akt pathway[J]. Int J Biol Sci, 2012, 8(1): 1-14. DOI: 10.7150/ijbs.8.1.
 

更新日期/Last Update: 2019-05-24