吡格列酮对K562/ADR细胞多药耐药的逆转作用及机制
|
摘要
目的:探讨吡格列酮对K562/ADR细胞阿霉素耐药的逆转作用及机制。方法:应用MTT法检测吡格列酮对K562及K562/ADR细胞的增殖抑制率、半数抑制浓度(IC50)、耐药逆转倍数并绘制增殖抑制率曲线;应用RT-PCR法测定PPARγ、CYP2C8和CYP2J2基因的转录;Western blot法测定PPARγ、CYP2C8和CYP2J2蛋白的表达。结果:吡格列酮作用于K562及K562/ADR细胞60 h的IC50分别为326μmol/L、349μmol/L;30μmol/L吡格列酮逆转K562/ADR细胞的阿霉素耐药倍数为6.4倍。吡格列酮作用于K562/ADR细胞后,CYP2C8和CYP2J2基因转录明显下降,CYP2C8、CYP2J2蛋白表达明显降低,PPARγ转录和PPARγ蛋白表达无差异。结论:吡格列酮可逆转K562/ADR细胞对阿霉素的耐药,其机制与CYP2C8和CYP2J2蛋白下调密切相关。吡格列酮是一种有效的肿瘤多药耐药逆转剂。
Objective: To explore the reversal effect of pioglitazone(PIO) on multidrug resistance in K562/ADR cells and its mechanism. Methods: The proliferation inhibition rate, half inhibition concentration(IC_(50)) and drug-resistance reversal multipe were detected and the curve of proliferation inhibition rate was drawn by MTT assay, the transcription of PPARy, CYP2C8 and CYP2J2 genes was detected by RT-PCR; the expression of PPARγ, CYP2C8 and CYP2J2 proteins was detected by Western blot. Results: The IC_(50) of PIO on K562 and K562/ADR cells for 60 h was 326.7 μmol/L and 349.1 μmol/L respectively. The reversal multiple of 30 μmol/L PIO on ADR-resistance of K562/ADR cells was6.4. After treatment of K562/ADR cells with PIO, the transcription of CYP2C8 and CYP2J2 and the protein expression of CYP2C8 and CYP2J2 significantly decreased, the transcription of PPARγ gene and the expression of PPARγ protein were not changed. Conclusions: Pioglitazone can reverse the adriamycin-resistance in K562/ADR cells that is closely related to the decrease of protein expression of CYP2C8 and CYP2J2. Pioglitazone is an effective multidrug resistance reversal agent for tumors.
Objective: To explore the reversal effect of pioglitazone(PIO) on multidrug resistance in K562/ADR cells and its mechanism. Methods: The proliferation inhibition rate, half inhibition concentration(IC_(50)) and drug-resistance reversal multipe were detected and the curve of proliferation inhibition rate was drawn by MTT assay, the transcription of PPARy, CYP2C8 and CYP2J2 genes was detected by RT-PCR; the expression of PPARγ, CYP2C8 and CYP2J2 proteins was detected by Western blot. Results: The IC_(50) of PIO on K562 and K562/ADR cells for 60 h was 326.7 μmol/L and 349.1 μmol/L respectively. The reversal multiple of 30 μmol/L PIO on ADR-resistance of K562/ADR cells was6.4. After treatment of K562/ADR cells with PIO, the transcription of CYP2C8 and CYP2J2 and the protein expression of CYP2C8 and CYP2J2 significantly decreased, the transcription of PPARγ gene and the expression of PPARγ protein were not changed. Conclusions: Pioglitazone can reverse the adriamycin-resistance in K562/ADR cells that is closely related to the decrease of protein expression of CYP2C8 and CYP2J2. Pioglitazone is an effective multidrug resistance reversal agent for tumors.
引文
1 Perez-Herrero E,Fernandez-Medarde A.Advanced targeted therapies in cancer:drug nanocarriers,the future of chemotherapy.Eur J Pharm Biopharm,2015;93:52-79.
2 Beretta GL,Cavalieri F.Engineering nanomedicines to overcome multidrug resistance in cancer therapy.Curr Med Chem,2016;23(1):3-22.
3 Nelson DR.The cytochrome P450 homepage.Human Genomics,2009,4(1):59-65.
4 Rudolf JD,Chang CY,Ma M,et al.Cytochromes P450 for natural product biosynthesis in Streptomyces:sequence,structure,and function.Nat Prod Rep,2017;34(9):1141-1172.
5 Zhang X,Li S.Expansion of chemical space for natural products by uncommon P450 reactions.Nat Prod Rep,2017;34(9):1061-1089.
6 Scripture CD,Sparreboom A,Figg WD.Modulation of cytochrome P450 activity:implications for cancer therapy.Lancet Oncol,2005;6(10):780-789.
7 Lee CA,Neul D,Clouser-Roche A,et al.Identification of novel substrates for human cytochrome P450 2J2.Drug Metab Dispos,2010;38(2):347-356.
8 Enayetallah AE,French RA,Thibodeau MS,et al.Distribution of soluble epoxide hydrolase and of cytochrome P450 2C8,2C9,and2J2 in human tissues.J Histochem Cytochem,2004;52(4):447-454.
9 Bieche I,Narjoz C,Asselah T,et al.Reverse transcriptase-PCRquantification of mRNA levels from cytochrome(CYP)1,CYP2and CYP3 families in 22 different human tissues.Pharmacogenet Genomics,2007;17(9):731-742.
10 Kota BP,Huang TH,Roufogalis BD.An overview on biological mechanisms of PPARs.Pharmacol Res,2005;51(2):85-94.
11 Gou Q,Gong X,Jin J,et al.Peroxisome proliferator-activated receptors(PPARs)are potential drug targets for cancer therapy.Oncotarget,2017;8(36):60704-60709.
12 Rakhshandehroo M,Knoch B,Muller M,et al.Peroxisome proliferator-activated receptor alpha target genes.PPAR Res,2010;2010.doi:10.1155/2010/612089.
13 Sahi J,Black CB,Hamilton GA,et al.Comparative effects of thiazolidinediones on in vitro P450 enzyme induction and inhibition.Drug Metab Dispos,2003;31(4):439-446.
14 Denisov IG,Frank DJ,Sligar SG.Cooperative properties of cytochromes P450.Pharmacol Ther,2009;124(2):151-167.
2 Beretta GL,Cavalieri F.Engineering nanomedicines to overcome multidrug resistance in cancer therapy.Curr Med Chem,2016;23(1):3-22.
3 Nelson DR.The cytochrome P450 homepage.Human Genomics,2009,4(1):59-65.
4 Rudolf JD,Chang CY,Ma M,et al.Cytochromes P450 for natural product biosynthesis in Streptomyces:sequence,structure,and function.Nat Prod Rep,2017;34(9):1141-1172.
5 Zhang X,Li S.Expansion of chemical space for natural products by uncommon P450 reactions.Nat Prod Rep,2017;34(9):1061-1089.
6 Scripture CD,Sparreboom A,Figg WD.Modulation of cytochrome P450 activity:implications for cancer therapy.Lancet Oncol,2005;6(10):780-789.
7 Lee CA,Neul D,Clouser-Roche A,et al.Identification of novel substrates for human cytochrome P450 2J2.Drug Metab Dispos,2010;38(2):347-356.
8 Enayetallah AE,French RA,Thibodeau MS,et al.Distribution of soluble epoxide hydrolase and of cytochrome P450 2C8,2C9,and2J2 in human tissues.J Histochem Cytochem,2004;52(4):447-454.
9 Bieche I,Narjoz C,Asselah T,et al.Reverse transcriptase-PCRquantification of mRNA levels from cytochrome(CYP)1,CYP2and CYP3 families in 22 different human tissues.Pharmacogenet Genomics,2007;17(9):731-742.
10 Kota BP,Huang TH,Roufogalis BD.An overview on biological mechanisms of PPARs.Pharmacol Res,2005;51(2):85-94.
11 Gou Q,Gong X,Jin J,et al.Peroxisome proliferator-activated receptors(PPARs)are potential drug targets for cancer therapy.Oncotarget,2017;8(36):60704-60709.
12 Rakhshandehroo M,Knoch B,Muller M,et al.Peroxisome proliferator-activated receptor alpha target genes.PPAR Res,2010;2010.doi:10.1155/2010/612089.
13 Sahi J,Black CB,Hamilton GA,et al.Comparative effects of thiazolidinediones on in vitro P450 enzyme induction and inhibition.Drug Metab Dispos,2003;31(4):439-446.
14 Denisov IG,Frank DJ,Sligar SG.Cooperative properties of cytochromes P450.Pharmacol Ther,2009;124(2):151-167.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |