米尔贝逆转耐阿霉素人乳腺癌细胞多药耐药的作用
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摘要
目的探寻米尔贝类化合物尼莫克汀、米尔贝β1逆转人乳腺癌多药耐药细胞株(MCF-7/adr)多药耐药(multidrug resistance,MDR)的作用及机制。方法采用MTT比色法测定细胞生长抑制率及耐药指数;高效液相色谱(HPLC)检测细胞内阿霉素(ADR)的积累变化;荧光分光光度仪检测罗丹明123(Rh123)在肿瘤细胞内的积累;通过RT-PCR与流式细胞仪检测MDR1基因与P-糖蛋白(P-gp)表达的变化。结果 5μmol·L-1的尼莫克汀、米尔贝β1可明显增强MCF-7/adr对ADR的敏感性,增加细胞内ADR及Rh123的积累,且呈剂量依赖关系,不同程度降低MDR1和P-gp的表达。结论尼莫克汀、米尔贝β1对MCF-7/adr的耐药有一定的逆转作用,且尼莫克汀效果好于米尔贝β1。
Aim To explore the reversal of MDR(multidrug resistance) by nemadectin and milbemycin β1 from the milbemycin family to adriamycin-resistant human breast carcinoma cells(MCF-7 / adr). Methods MTT assay was used to evaluate the cell suppression rates and reversal effects. Intracellular accumulation of adriamycin was determined by HPLC. The accumulation of rhodamine 123(Rh123) was measured with spectrofluorometer. The expressions of P-gp and the MDR1 gene encoding P-gp were tested by flow cytometry and reverse-transcriptase PCR. Results Nemadectin and milbemycin β1at the high dose of 5 μmol·L- 1significantly increased the sensitivity of MCF-7 / adr cells to adriamycin. They also increased the intracellular accumulation of adriamycin and Rh123 in MCF-7 /adr cells in a dose-dependent manner. Expression of both P-gp and MDR1 was down-regulated to different level. Conclusion Nemadectin and milbemycin β1can partly reverse multidrug resistance of MCF-7 / adr,and the effect of nemadectin is better than that of milbemycin β1.
Aim To explore the reversal of MDR(multidrug resistance) by nemadectin and milbemycin β1 from the milbemycin family to adriamycin-resistant human breast carcinoma cells(MCF-7 / adr). Methods MTT assay was used to evaluate the cell suppression rates and reversal effects. Intracellular accumulation of adriamycin was determined by HPLC. The accumulation of rhodamine 123(Rh123) was measured with spectrofluorometer. The expressions of P-gp and the MDR1 gene encoding P-gp were tested by flow cytometry and reverse-transcriptase PCR. Results Nemadectin and milbemycin β1at the high dose of 5 μmol·L- 1significantly increased the sensitivity of MCF-7 / adr cells to adriamycin. They also increased the intracellular accumulation of adriamycin and Rh123 in MCF-7 /adr cells in a dose-dependent manner. Expression of both P-gp and MDR1 was down-regulated to different level. Conclusion Nemadectin and milbemycin β1can partly reverse multidrug resistance of MCF-7 / adr,and the effect of nemadectin is better than that of milbemycin β1.
引文
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[11]陈淑娴,朱瑞宇,付强,等.人乳腺癌细胞及其耐药细胞转录因子表达的差异[J].中国药理学通报,2012,28(9):1257-61.[11]Chen S X,Zhu R Y,Fu Q,et al.The transcription differences ofmammary breast cancer cell and its adriamycin-resistant cells[J].Chin Pharmacol Bull,2012,28(9):1257-61.
[2]戴春岭,符立梧.肿瘤多药耐药逆转剂的研究进展[J].中国药理学通报,2005,21(5):513-8.[2]Dai C L,Fu L W.The development of the reversal of the tumormultidrug resistance[J].Chin Pharmacol Bull,2005,21(5):513-8.
[3]葛涵,沈舜义.大环内酯类抗生素研究进展[J].世界临床药物,2007,28(6):376-80.[3]Ge H,Shen S Y.The advances of the research on macrolide anti-biotics[J].World Clin Drugs,2007,28(6):376-80.
[4]王建强,马淑涛.16元大环内酯类抗生素的研究进展[J].中国药学杂志,2006,41(24):1841-4.[4]Wang J Q,Ma S T.The advances of the research on 16-macrolideantibiotics[J].Chin Pharmaceut J,2006,41(24):1841-4.
[5]Korystov Y N,Ermakova N V,Kublik L N,et al.Avermectinsinhibit multidrug resistance of tumor cells[J].Eur J Pharmacol,2004,493(1-3):57-64.
[6]Chen L M,Liang Y J,Ruan J W,et al.Reversal of P-gp media-ted multidrug resistance in-vitro and in-vivo by FG020318[J].JPharm Pharmacol,2004,56(8):1061-6.
[7]Shi Y Q,Qu X J,Liao Y X,et al.Reversal effect of a macrocy-clic bisbibenzyl plagiochin E on multidrug resistance in adriamy-cin-resistant K562/A02 cells[J].Eur J Pharmacol,2008,584(1):66-71.
[8]Pouliot J F,L'Heureux F,Liu Z,et al.Reversal of P-glycopro-tein-associated multidrug resistance by ivermectin[J].BiochemPharmacol,1997,53(1):17-25.
[9]Tsukiyama T,Kinoshita A,Ichinose R,et al.Synthesis of milbe-mycins b 9 and b10 from milbemycins A3 and A4 and their biolog-ical activities[J].Biosci Biotechnol Biochem,2002,66(6):1407-11.
[10]Tsukiyama T,Kajino H,Tsukamoto Y,et al.Synthesis of novel26-substituted milbemycin A4 derivatives and their acaricidal ac-tivities[J].J Antibiot,2004,57(7):446-55.
[11]陈淑娴,朱瑞宇,付强,等.人乳腺癌细胞及其耐药细胞转录因子表达的差异[J].中国药理学通报,2012,28(9):1257-61.[11]Chen S X,Zhu R Y,Fu Q,et al.The transcription differences ofmammary breast cancer cell and its adriamycin-resistant cells[J].Chin Pharmacol Bull,2012,28(9):1257-61.