PFC体内递增诱导建立S180肿瘤多药耐药模型及其稳定性观察

doi:10.3969/j.issn.1674-5817.2015.05.005

实验动物与比较医学 ›› 2015, Vol. 35 ›› Issue (5): 367-373.DOI: 10.3969/j.issn.1674-5817.2015.05.005

PFC体内递增诱导建立S180肿瘤多药耐药模型及其稳定性观察

顾云浩, 曹晨洁, 胡碧原, 王俊, 韩东冬, 许爱华

扬州大学医学院, 扬州 225001

收稿日期:2015-03-03 出版日期:2015-10-25 发布日期:2015-10-25
作者简介:顾云浩(1994-), 男, 扬州大学医学院2012级在读本科生。E-mail: 907745184@qq.com
基金资助:
扬州大学大学生科技创新基金项目(2014); 江苏省医药高技术研究项目(BG2007609)

Establishment of S180 Tumor Multidrug Resistance Mouse Model by Increasing PFC and Observation on Stability

GU Yun-hao, CAO Chen-jie, HU Bi-yuan, WANG Jun, HAN Dong-dong, XU Ai-hua

Medical College, Yangzhou University, Yangzhou 225001, China

Received:2015-03-03 Online:2015-10-25 Published:2015-10-25

摘要/Abstract

摘要： 目的小鼠体内诱导建立获得性S180多药耐药(multi-drug resistance, MDR)模型及其稳定性观察。方法模拟临床顺铂+氟尿嘧啶+环磷酰胺(PFC)化疗方案给药, 分三个阶段剂量递增法诱导S180腹水瘤小鼠, 建立获得性S180MDR实验模型。采用噻唑蓝(MTT)法、流式细胞术动态检测各阶段所诱导细胞对化疗药物的耐药倍数、细胞内药物积累量及细胞膜P-糖蛋白(P-glycoprotein, P-gp)功能活性,并通过检测以上指标观察各阶段所诱导细胞停药后的耐药稳定性; 采用实时荧光定量PCR(RT-qPCR)法检测各阶段所诱导细胞MDR-1 mRNA、多药耐药相关蛋白-1(multidrug resistance-associated protein 1,MRP-1) mRNA的表达量。结果与亲本细胞对照组比较,各阶段所诱导S180细胞对化疗药物的耐药倍数随着诱导时间延长和剂量增高而逐渐增大, 细胞内阿霉素(adriamycin, ADR)积累量逐渐减少, 细胞P-gp功能活性逐渐增强; 各阶段所诱导S180细胞MDR-1 mRNA、MRP-1 mRNA的表达量也与诱导时间和给药剂量呈正相关; 第一、二和三阶段所诱导细胞的稳定耐药时间分别为1周、2周和3周左右。结论模拟临床PFC化疗方案给药,采用分阶段剂量递增小鼠体内诱导法可建立耐药强度高、稳定时间长的获得性S180MDR实验模型。

关键词: 顺铂+氟尿嘧啶+环磷酰胺(PFC), 多药耐药(MDR), S180细胞株, 体内, 细胞膜P-糖蛋白(P-gp), MDR-1 mRNA, 多药耐药相关蛋白-1(MRP-1) mRNA

Abstract: Objective To establish aquired S180 multidrug resistance (MDR) mouse model and study on its stability. Methods To mimic the clinical PFC (cis-Dichlorodiamineplatinum+5-Fluorouracil+cyclophosphamide) scheme, gradually increase the dose in three phases to induce S180 ascites tumor mice and establish the aquired S180 MDR mouse model. The induced cells resistance factor to the chemotherapeutics drugs in different stages, the accumulation of adriamycin (ADR) and the functional activity of P-glycoprotein (P-gp) were detected by MTT assay and flow cytometry. And then through detecting the above indicators to monitor the resistance drug stability of induced cells in different stages. The mRNA expression of MDR-1 and multidrug resistance-associated protein 1 (MRP-1) of induced cells in different stages were detected by real time quantitative PCR (RT-qPCR). Results Compared with the parent cells, with induction time extending and dose increasing, the resistance factors in each stage of induced S180 cells to chemotherapeutics drugs were gradually increased, the accumulation of ADR was gradually reduced, and the functional activity of P-gp was strengthened. The mRNA expression of MDR-1 and MRP-1 of induced cells in different stages had a positive correlation to the induction time and dose. The stable resistance time of induced cells in the first, second and third phase are respectively for about 1 week, 2 weeks and 3 weeks. Conclusion To mimic the clinical PFC scheme, using the dose gradually increasing by phased can establish a high resistant strength, long stable time aquired S180MDR experimental model.

Key words: cis-Dichlorodiamineplatinum+5-Fluorouracil+cyclophosphamide (PFC), Multidrug resistance, S180 cell line, in vivo, P-gp, multidrug resistance-1(MDR-1mRNA), multidrug resistance-associated protein-1(MRP-1 mRNA)

中图分类号:

Q95-33

顾云浩, 曹晨洁, 胡碧原, 王俊, 韩东冬, 许爱华. PFC体内递增诱导建立S180肿瘤多药耐药模型及其稳定性观察[J]. 实验动物与比较医学, 2015, 35(5): 367-373.

GU Yun-hao, CAO Chen-jie, HU Bi-yuan, WANG Jun, HAN Dong-dong, XU Ai-hua. Establishment of S180 Tumor Multidrug Resistance Mouse Model by Increasing PFC and Observation on Stability[J]. Laboratory Animal and Comparative Medicine, 2015, 35(5): 367-373.

参考文献

[1] Huang F, Wu XN, Chen J, et al.Resveratrol reverses multidrug resistance in human breast cancer doxorubicin-resistant cells[J]. Exp Ther Med, 2014.7:1611-1616.
[2] Zhang QY, Wang JZ, He HB, et al.Trametenolic acid B reverses multidrug resistance in breast cancer cells through regulating the expression level of P-Glycoprotein[J]. Phytother Res,2014, 28:1037-1044.
[3] 刘晓冬, 常青, 赵秀芳, 等. 多西他赛诱导肺腺癌A549/DTX细胞株的建立及耐药机制分析[J]. 肿瘤防治研究, 2014, 41(6):519-522.
[4] Tian QE, Li HD, Yan M, et al.Effects of astragalus polysaccharides on P-glycoprotein efflux pump function and protein expression in H22 hepatoma cells in vitro[J]. BMC Complement Altern Med, 2012, 12:94.
[5] Zhou Y, Ling XL, Li SW, et al.Establishment of a human hepatoma multidrug resistant cell line in vitro[J]. World J Gastroenterol, 2010, 16(18):2291-2297.
[6] Li GY, Liu JZ, Zhang B, et al.Tegillarca granosa extract Haishengsu (HSS) suppresses expression of mdr1, BCR/ABL and sorcin in drug-resistant K562/ADR tumors in mice[J]. Adv Med Sci, 2013, 58(1):112-117.
[7] 黄永明, 苏海涛, 黄启明, 等. 人骨肉瘤耐药性KH-OS/ADR细胞SCID小鼠移植模型的建立及其生物学特征的研究[J]. 中国中医骨伤科杂志, 2013, 21(7):1-3.
[8] Zrieki A, Farinotti R, Buyse M.Cyclooxygenase-2 inhibitors prevent trinitrobenzene sulfonic acid-induced P-glycoprotein up-regulation in vitro and in vivo[J]. Eur J Pharmacol,2010, 636(1-3):189-197.
[9] Loetchutinat C, Saengkhae C, Marbeuf-Gueye C, et al.New insights into the P-glycoprotein-mediated effluxes of rhodamines[J]. Eur J Biochem, 2003, 270(3):476-485.
[10] Snow K, Judd W.Characteristion of adriamycin and amsacrine-resistant human leukaemic T cell lines[J]. Br J Cancer, 1991,63(1):17-28.
[11] 徐龙进, 王祖海, 孙付军, 等. 联合化疗诱导小鼠在体肿瘤多药耐药模型的建立[J]. 实验动物与比较医学, 2005, 25(4):215-217.
[12] 陈强, 李贵海, 孙付军, 等. 适合中药研究的小鼠S180肿瘤获得性多药耐药模型的建立及稳定性研究[J].中草药, 2006, 37(11):1691-1694.
[13] 尹格平, 顾清, 陈铭, 等. 腹水型S180细胞系获得性MDR表达鼠模型的建立[J]. 上海免疫学杂志, 2001, 21(5):282-285.
[14] Juranka PF, Zastawny RL, Ling V.P-glycoprotein: multidrug-resistance and a superfamily of membrane-associated transport proteins[J]. FASEB J, 1989, 3(14):2583-2592.
[15] Molinas A, Sicard G, Jakob I.Functional evidence of multidrug resistance transporters (MDR) in rodent olfactory epithelium[J]. PLoS One, 2012, 7(5):1-13.
[16] Hennessy M, Spiers JP.A primer on the mechanics of P-glycoprotein the multidrug transporter[J]. Pharmacol Res, 2007, 55(1):1-15.
[17] Villar VH, V gler O, BarcelóF, et al. Oleanolic and maslinic acid sensitize soft tissue sarcoma cells to doxorubicin by inhibiting the multidrug resistance protein MRP-1, but not P-glycoprotein[J]. J Nutr Biochem, 2014, 25:429-438.

PFC体内递增诱导建立S180肿瘤多药耐药模型及其稳定性观察

Establishment of S180 Tumor Multidrug Resistance Mouse Model by Increasing PFC and Observation on Stability

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 6

编辑推荐

Metrics

本文评价

[1]	李洪波, 黄锐, 代将, 钟振东. 竹节香附素A对肝缺血再灌注大鼠的保护作用机制[J]. 实验动物与比较医学, 2021, 41(1): 55-60.
[2]	匡德宣, 王文广, 孙晓梅, 代解杰. 恶性疟原虫动物模型及基因编辑研究进展[J]. 实验动物与比较医学, 2019, 39(1): 65-71.
[3]	彭礼繁, 张小建, 廖梅旭, 李晓洁, 余鲲. 超促排卵后不同取卵时间对昆明小鼠卵子质量、体内成熟率和体外受精率的影响[J]. 实验动物与比较医学, 2017, 37(5): 405-409.
[4]	邵宝珠, 唐其凤, 曹丽丽. BALB/c小鼠不同体质量范围对重组人促红素注射液体内活性测定的影响[J]. 实验动物与比较医学, 2016, 36(3): 216-218.
[5]	游根娣1, 蒋秀蓉2, 张燕林2. 小鼠卵在体内外受精和胚胎发育的比较[J]. , 1997, 17(4): 237-238.
[6]	许兰文，陈绍亮，杨幼明，杨俊华，赵为之，汤钊猷，钟高仁，朱桐. ¹³¹I标记PMT的代谢及毒性试验结果[J]. , 1993, 13(2): 75-77.