[1]高 峰,王秀会,夏胜利,等.雷公藤甲素对人骨肉瘤细胞阿霉素耐药株的逆转作用及机制研究[J].陕西医学杂志,2021,50(6):648-651,656.[doi:DOI:10.3969/j.issn.1000-7377.2021.06.002]
　GAO Feng,WANG Xiuhui,XIA Shengli,et al.Reversal effect and mechanism of triptolide on adriamycin-resistant human osteosarcoma cell line[J].,2021,50(6):648-651,656.[doi:DOI:10.3969/j.issn.1000-7377.2021.06.002]
点击复制

雷公藤甲素对人骨肉瘤细胞阿霉素耐药株的逆转作用及机制研究
分享到：

《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
50
期数:
2021年6期
页码:
648-651,656
栏目:
基础研究
出版日期:
2021-06-05

文章信息/Info

Title:
Reversal effect and mechanism of triptolide on adriamycin-resistant human osteosarcoma cell line
作者:
高 峰; 王秀会; 夏胜利; 周小小; 易存国; 付备刚; 沈 超; 王明辉
(上海健康医学院附属周浦医院 上海市浦东新区周浦医院,上海 201318)
Author(s):
GAO Feng; WANG Xiuhui; XIA Shengli; ZHOU Xiaoxiao; YI Cunguo; FU Beigang; SHEN Chao; WANG Minghui
(Zhoupu Hospital Affiliated to Shanghai Medical College,Shanghai 201318,China)
关键词:
雷公藤甲素;  多药耐药基因;  人骨肉瘤细胞阿霉素耐药株;  PI3K通路;  P-糖蛋白;  肺耐药相关蛋白
Keywords:
Triptolide;  Multidrug resistance gene;  Adriamycin-resistant human osteosarcoma cell line;  PI3K pathway;  P-glycoprotein;  Lung resistance-related protein
分类号:
R 738.1
DOI:
DOI:10.3969/j.issn.1000-7377.2021.06.002
文献标志码:
A
摘要:
目的:探讨雷公藤甲素(TPL)体外给药对人骨肉瘤细胞阿霉素(ADR)耐药株(U2OS/ADR)多药耐药(MDR)的逆转作用及可能机制。方法:使用人骨肉瘤细胞株U2OS,利用浓度递增法建立耐药骨肉瘤细胞U2OS/ADR模型。设置空白组、对照组、TPL组和维拉帕米阳性对照组(VER组)。CCK-8法测定TPL处理对U2OS/ADR细胞增殖和ADR敏感性的影响。以罗丹明123(Rh123)作为内标,采用流式细胞术评估细胞P-糖蛋白(P-gp)外排作用。实时定量PCR(qRT-PCR)检测TPL对MDR1基因转录的影响。Western blot检测TPL对磷酸化磷酸肌醇-3-激酶(p-PI3K)、磷酸化蛋白激酶B(p-AKT)、MDR1和肺耐药相关蛋白(LRP)表达的影响。结果:与对照组相比,TPL组使ADR对U2OS/ADR细胞生长的抑制作用提高(P<0.05)。TPL组Rh123阳性信号计数高于对照组(P<0.001)。与对照组相比,TPL组MDR1、LRP、p-PI3K和p-AKT蛋白表达水平及MDR1基因mRNA水平下降(均P<0.05)。结论:TPL具有抑制U2OS/ADR细胞生长、提高U2OS对ADR敏感性的作用,其机制可能与抑制PI3K/AKT信号通路活化、下调U2OS/ADR细胞MDR1和LRP基因表达、抑制P-gp对细胞毒物质的外排有关。
Abstract:
Objective:To explore the reversal effect and possible mechanism of triptolide(TPL)in vitro on the multidrug resistance(MDR)of human osteosarcoma cell lines resistant to adriamycin(ADR)(U2OS/ADR).Methods:U2OS was used to establish a drug-resistant osteosarcoma cell model U2OS/ADR by increasing concentration scheme.The blank group,control group,TPL group and verapamil positive control group(VER group)were set up.CCK-8 test was used to determine the effects of TPL treatment on U2OS/ADR cell proliferation and ADR sensitivity.Rhodamine 123(Rh123)was used as an internal standard,and flow cytometry was used to evaluate cellular P-glycoprotein(P-gp)efflux.Real-time quantitative PCR(qRT-PCR)was used to detect the effect of TPL on the transcription of MDR1.Western blot was used to detect the effects of TPL on p-PI3K,p-AKT,MDR1 and LRP expression levels.Results:Compared with the control group,the TPL group increased the inhibitory effect of ADR on U2OS/ADR cell growth(P<0.05).The positive signal count of Rh123 in the TPL group was higher than that in the control group(P<0.001).Compared with the control group,the protein expression levels of MDR1,LRP,p-PI3K and p-AKT and the mRNA level of MDR1 gene decreased in the TPL group(all P<0.05).Conclusion:TPL can inhibit the growth of U2OS/ADR cells and increase the sensitivity of U2OS to ADR.The mechanism may be related to the inhibition of PI3K/AKT signal pathway activation,down-regulation of MDR1 and LRP gene expression in U2OS/ADR cells,and inhibition of the efflux of cytotoxic substances by P-gp.

参考文献/References:

[1] 卢文婷,刘 爽,李赛赛,等.FHL2沉默对骨肉瘤U2OS细胞增殖与凋亡的影响[J].临床肿瘤学杂志,2018,23(1):1-6.
[2] 王海娟,张美芝,刘小菊,等.雷公藤内酯醇逆转肿瘤多药耐药研究进展[J].中国实验方剂学杂志,2016,22(10):229-234.
[3] Ma P,Dong X,Swadley CL,et al.Development of idarubicin and doxorubicin solid lipid nanoparticles to overcome Pgp-mediated multiple drug resistance in leukemia[J].J Biomed Nanotechnol,2009,5(2):151-161.
[4] 张 倩,彭广操,朱明军.雷公藤的药理作用及毒性研究进展[J].中西医结合心脑血管病杂志,2016,(15):1753-1754.
[5] 崔 进,陈 晓,苏佳灿.雷公藤甲素药理作用研究新进展[J].中国中药杂志,2017,42(14):2655-2658.
[6] 蔡灯塔,王新春,王景春.雷公藤甲素联合来氟米特治疗类风湿关节炎的临床效果及对患者血清炎性因子的影响[J].陕西中医,2018,39(9):1260-1262.
[7] 刘 倩,汤建华,张志华.雷公藤甲素对非小细胞肺癌细胞系A549增殖的抑制和凋亡的诱导[J].基础医学与临床,2020,40(3):315-320.
[8] 惠璐璐,许文林,陈巧云,等.雷公藤甲素对K562/A02细胞阿霉素敏感性的影响[J].中国实验血液学杂志,2012,20(1):66-69.
[9] 陈 军,金雪荣,王 翔,等.shRNA干扰COPS3基因对人成骨肉瘤细胞的影响[J].陕西医学杂志,2017,46(3):278-281.
[10] Lourda M,Trougakos IP,Gonos ES.Development of resistance to chemotherapeutic drugs in human osteosarcoma cell lines largely depends on upregulation of clusterin/Apolipoprotein J [J].Int J Cancer,2007,120(3):611-622.
[11] Takara K,Sakaeda T,Okumura K.An update on overcoming MDR-1 mediated multidrug resistance in cancer chemotherapy[J].Curr Pharm Des,2006,12(3):273-286.
[12] 贵 鹏,李朝旭,吴 昊,等.MiRNA靶向调控骨肉瘤耐药机制的研究进展[J].中国肿瘤,2019,28(9):694-698.
[13] Celia M,Freitas G,Heidi VP,et al.Mutidrug resistance mediated by ABC transporters in osteosarcoma cell lines:MRNA ananlysis and functional radiotracer studies[J].Nucl Med Biol,2006,33:831-840.
[14] 袁玉霞,殷佩浩.中药单体逆转MDR1介导的肿瘤多药耐药机制研究进展[J].上海中医药杂志,2017,51(3):96-99.
[15] 汤 涛,董 伟,张 婧,等.中药单体成分逆转肿瘤多药耐药的研究进展[J].中草药,2017,48(4):792-797,846.
[16] 周炳刚,魏昌晟,张 颂,等.苦参碱逆转人乳腺癌MCF-7/ADR细胞株耐药性及对PI3K/AKT信号通路下游因子的影响[J].现代肿瘤医学,2017,25(1):9-13.
[17] Marsousi N,Doffey-Lazeyras F,Rudaz S,et al.Intestinal permeability and P-glycoprotein-mediated efflux transport of ticagrelor in Caco-2 monolayer cells[J].Fundam Clin Pharmacol,2016,30(6):577-584.
[18] 刘风玲,杨 旭.粉防己碱逆转人卵巢癌耐药细胞SKOV3/DDP耐药的逆转作用及机制[J].山东医药,2011,51(45):37-38.
[19] Chen LM,Song TJ,Xiao JH,et al.Tripchlorolide induces autophagy in lung cancer cells by inhibiting the PI3K/AKT/mTOR pathway and improves cisplatin sensitivity in A549/DDP cells[J].Oncotarget,2017,8(38):63911-63922.
[20] Li H,Hui L,Xu W,et al.Modulation of P-glycoprotein expression by triptolide in adriamycin-resistant K562/A02 cells[J].Oncol Lett,2012,3(2):485-489.

相似文献/References:

[1]闫 真,王雅玲,高 枫.雷公藤甲素对脂多糖诱导脊髓损伤小鼠及BV2小胶质细胞的调节作用及机制研究[J].陕西医学杂志,2021,50(6):643.[doi:DOI:10.3969/j.issn.1000-7377.2021.06.001]
　YAN Zhen,WANG Yaling,GAO Feng.Regulation effect and mechanism of triptolide on lipopolysaccharide-induced spinal cord injury in mice and BV2 microglia[J].,2021,50(6):643.[doi:DOI:10.3969/j.issn.1000-7377.2021.06.001]

备注/Memo

备注/Memo:
基金项目:上海市科学技术委员会科研计划项目(16ZR1431600); 上海市浦东新区卫生健康委员会临床特色学科建设计划项目(PWYTS2018-02); 上海健康医学院师资人才百人库项目(IPBRK-18-07)
更新日期/Last Update: 2021-06-07