榄香烯乳对人骨肉瘤细胞系（MG-63）多药耐药的逆转及与谷胱甘肽S-转移酶-π（GST-π）表达关系的体外研究
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摘要
目的:研究体外实验中榄香烯对人骨肉瘤细胞系(MG-63)多药耐药的逆转效果,探讨与GST-π的关系。
方法:采用MTT法测定阿霉素对人骨肉瘤细胞系(MG-63)的半数抑制浓度(IC_(50))以及榄香烯对人骨肉瘤细胞系(MG-63)的低细胞毒浓度(IC_(15))和非细胞毒浓度(IC_(05))。将细胞分为A、B、C三组,A组为对照组,加入半数抑制浓度的阿霉素;B、C组在A组基础上还加入非细胞毒浓度、低细胞毒浓度的榄香烯,细胞传代培养至A组中MG-63细胞对半数抑制浓度的阿霉素完全耐药后停止干预,检测以下指标:①阿霉素对干预后各组MG-63细胞的半数抑制浓度,计算耐药倍数(RF);②采用荧光分光光度计测定各组OD值,计算细胞内阿霉素药物浓度;③免疫组化法测定原始MG-63细胞和A、B、C组MG-63细胞谷胱甘肽S-转移酶-π(GST-π)的表达,并对以上数据进行分析比较各处理组间的差异。
结果:①阿霉素对干预后A、B、C组MG-63细胞的半数抑制浓度分别为3.37μg/ml、2.21μg/ml、1.19μg/ml,A、B、C组MG-63细胞对阿霉素的耐药倍数(RF)分别为11.62、7.62和4.10,A组＞B组＞C组,差异有统计学意义。②干预后A、B、C组MG-63细胞内阿霉素药物浓度分别为1.21μg/10~6细胞、1.96μg/10~6细胞、2.71μg/10~6细胞,A组＜B组＜C组,差异有统计学意义。③细胞GSTs的表达率:原始MG-63细胞为10.25%、A组为35.75%、B组为25.25%、C组为20.5%,A组＞B组＞C组＞原始MG-63细胞,差异有统计学意义。
结论:①证实在体外运用非细胞毒浓度或低细胞毒浓度的榄香烯乳能有效抑制阿霉素所诱导的人骨肉瘤细胞MG-63的多药耐药性(MDR)。②榄香烯体外能下调人骨肉瘤MG-63细胞GST-π的表达。③榄香烯乳逆转人骨肉瘤MG-63细胞产生多药耐药性(MDR)机制可能是通过抑制GST-π的过度表达,提高细胞内抗癌药的药物浓度而实现的。
Objective:To study the Reversal Effect of Elemene on Multidrug-resistant of human osteosarcoma cell line(MG-63/ADM) in vitro and investigated the relationship between GST-πand the effect.
Methods:To investigate the IC_(50) of adriamycin(ADM),IC_(05) and IC_(15) of elemene(ELE) to MG-63 human osteosarcoma cell by MTT assay. The MG-63 human osteosarcoma cell divide into three groups randomly (A、B、C Group).Group A is the control group,to establish a multi-drug resistant of human MG-63 cells by short-time pulse exposure of the cell to the IC_(50) of adriamycin.The cell of Group B、C are exposed both in the same dosage of ADM with the control group and IC_(05)(group B) or IC_(15) (group C)of elemene.The exposure continued until the cell of Group A can be cultured stably in the nutritive medium which contain IC_(50) of ADM,then drug exposure are stopped in all experiment Groups.we repeat to investigate the IC_(50) of adriamycin to all post-drug intervention MG-63 human osteosarcoma cells by MTT assay.And intracellular accumulation of ADM were determined by fluorospectrophotometery. Immunohistochemistry was applied to study the GST-πexpression of initial and post-drug intervention MG-63 cells.At the end,we analyse the experimental result by statistics.
Results:①The IC_(50) of adriamycin(ADM) to MG-63 cells were 3.37μg/ml(GroupA)、2.21μg/ml(GroupB)、1.19μg/ml(GroupC),and the RF values were 11.62(GroupA)、7.62(GroupB) and 4.10(GroupC),the comparisons among Group A,GroupB and Group C have statistical significance(P＜0.05).②The intracellular accumulation of ADM all experiment groups were 1.21μg/10~6 cells(Group A)、1.96μg/10~6 cells(GroupB)、2.71μg/10~6 cells(Group C),Group A＜GroupB＜Group C,the comparisons among Group A,GroupB and Group C have statistical significance(P＜0.05).③The GST-πexpression rates of initial and post-drug intervention MG-63 cells were 10.25%(initial cells), 35.75%(Group A),25.25%(Group B) and 20.50%(Group C),the comparisons among Group A,GroupB and Group C have statistical significance(P＜0.05)
Conclusion:Firstly,we conclude that the multi-drug resistance to ADM of human osteosacoma cell(MG-63/ADM) could be reversed by Elemene with non-toxic or low-toxic dosage;Secondly the expression of GST-πcould be regulated down by Elemene;The last,the increased intracellular accumulation of ADM and downregulation of GST-πwould contribute to the reversal mechanism of Elemene.
方法:采用MTT法测定阿霉素对人骨肉瘤细胞系(MG-63)的半数抑制浓度(IC_(50))以及榄香烯对人骨肉瘤细胞系(MG-63)的低细胞毒浓度(IC_(15))和非细胞毒浓度(IC_(05))。将细胞分为A、B、C三组,A组为对照组,加入半数抑制浓度的阿霉素;B、C组在A组基础上还加入非细胞毒浓度、低细胞毒浓度的榄香烯,细胞传代培养至A组中MG-63细胞对半数抑制浓度的阿霉素完全耐药后停止干预,检测以下指标:①阿霉素对干预后各组MG-63细胞的半数抑制浓度,计算耐药倍数(RF);②采用荧光分光光度计测定各组OD值,计算细胞内阿霉素药物浓度;③免疫组化法测定原始MG-63细胞和A、B、C组MG-63细胞谷胱甘肽S-转移酶-π(GST-π)的表达,并对以上数据进行分析比较各处理组间的差异。
结果:①阿霉素对干预后A、B、C组MG-63细胞的半数抑制浓度分别为3.37μg/ml、2.21μg/ml、1.19μg/ml,A、B、C组MG-63细胞对阿霉素的耐药倍数(RF)分别为11.62、7.62和4.10,A组＞B组＞C组,差异有统计学意义。②干预后A、B、C组MG-63细胞内阿霉素药物浓度分别为1.21μg/10~6细胞、1.96μg/10~6细胞、2.71μg/10~6细胞,A组＜B组＜C组,差异有统计学意义。③细胞GSTs的表达率:原始MG-63细胞为10.25%、A组为35.75%、B组为25.25%、C组为20.5%,A组＞B组＞C组＞原始MG-63细胞,差异有统计学意义。
结论:①证实在体外运用非细胞毒浓度或低细胞毒浓度的榄香烯乳能有效抑制阿霉素所诱导的人骨肉瘤细胞MG-63的多药耐药性(MDR)。②榄香烯体外能下调人骨肉瘤MG-63细胞GST-π的表达。③榄香烯乳逆转人骨肉瘤MG-63细胞产生多药耐药性(MDR)机制可能是通过抑制GST-π的过度表达,提高细胞内抗癌药的药物浓度而实现的。
Objective:To study the Reversal Effect of Elemene on Multidrug-resistant of human osteosarcoma cell line(MG-63/ADM) in vitro and investigated the relationship between GST-πand the effect.
Methods:To investigate the IC_(50) of adriamycin(ADM),IC_(05) and IC_(15) of elemene(ELE) to MG-63 human osteosarcoma cell by MTT assay. The MG-63 human osteosarcoma cell divide into three groups randomly (A、B、C Group).Group A is the control group,to establish a multi-drug resistant of human MG-63 cells by short-time pulse exposure of the cell to the IC_(50) of adriamycin.The cell of Group B、C are exposed both in the same dosage of ADM with the control group and IC_(05)(group B) or IC_(15) (group C)of elemene.The exposure continued until the cell of Group A can be cultured stably in the nutritive medium which contain IC_(50) of ADM,then drug exposure are stopped in all experiment Groups.we repeat to investigate the IC_(50) of adriamycin to all post-drug intervention MG-63 human osteosarcoma cells by MTT assay.And intracellular accumulation of ADM were determined by fluorospectrophotometery. Immunohistochemistry was applied to study the GST-πexpression of initial and post-drug intervention MG-63 cells.At the end,we analyse the experimental result by statistics.
Results:①The IC_(50) of adriamycin(ADM) to MG-63 cells were 3.37μg/ml(GroupA)、2.21μg/ml(GroupB)、1.19μg/ml(GroupC),and the RF values were 11.62(GroupA)、7.62(GroupB) and 4.10(GroupC),the comparisons among Group A,GroupB and Group C have statistical significance(P＜0.05).②The intracellular accumulation of ADM all experiment groups were 1.21μg/10~6 cells(Group A)、1.96μg/10~6 cells(GroupB)、2.71μg/10~6 cells(Group C),Group A＜GroupB＜Group C,the comparisons among Group A,GroupB and Group C have statistical significance(P＜0.05).③The GST-πexpression rates of initial and post-drug intervention MG-63 cells were 10.25%(initial cells), 35.75%(Group A),25.25%(Group B) and 20.50%(Group C),the comparisons among Group A,GroupB and Group C have statistical significance(P＜0.05)
Conclusion:Firstly,we conclude that the multi-drug resistance to ADM of human osteosacoma cell(MG-63/ADM) could be reversed by Elemene with non-toxic or low-toxic dosage;Secondly the expression of GST-πcould be regulated down by Elemene;The last,the increased intracellular accumulation of ADM and downregulation of GST-πwould contribute to the reversal mechanism of Elemene.
引文
[1] Cummings J, Ethell BT, Jardine L et-al. Glucuronidation as a mechanism of intrinsic drug resistance in human colon cancer: reversal of resistance by food additives. Cancer Res. 2003 ;63(23):8443-50.
[2] Barok M, Isola J, P(?)lyi-Krekk Z et-al.Trastuzumab causes antibody -dependent cellular cytotoxicity-mediated growth inhibition of submacroscopic JIMT-1 breast cancer xenografts despite intrinsic drug resistance. Mol Cancer Ther.2007 Jul;6(7):2065-72.
[3] De Anta JM, Perez-Castro AJ, Freire R et-al.The DNA damage checkpoint is activated during residual tumour cell survival to methotrexate treatment as an initial step of acquired drug resistance. Anticancer Drugs. 2006;17(10):1171-7
[4] Garc(?)a-Mart(?)n E, Pizarro RM, Martinez C et-al.Acquired resistance to the anticancer drug paclitaxel is associated with induction of cytochrome P450 2C8.Pharmacogenomics. 2006;7(4):575-85.
[5] Brozovic A, Fritz G, Christmann M et-al. Long-term activation of SAPK/JNK,p38 kinase and fas-L expression by cisplatin is attenuated in human carcinoma cells that acquired drug resistance. Int J Cancer. 2004; 112(6):974-85
[6] Friedrich K, Wieder T, Von Haefen C et-al. Overexpression of caspase-3 restores sensitivity for drug-induced apoptosis in breast cancer cell lines with acquired drug resistance. Oncogene. 2001;20(22):2749-60.
[7] Tian K, Jurukovski V, Wang XP et-al.Epigenetic regulation of WTH3 in primary and cultured drug-resistant breast cancer cells. Cancer Res.2005;65(21):10024-31
[8] Rellecke P, Kuchelmeister K, Schachenmayr W et-al.Mismatch repair protein hMSH2 in primary drug resistance in in vitro human malignant gliomas.J Neurosurg. 2004;101(4):653-8.
[9] Garcia R, Franklin RA, McCubrey JA et-al. EGF induces cell motility and multi-drug resistance gene expression in breast cancer cells.Cell Cycle.2006;5(23):2820-6.
[10] Mayur YC, Jagadeesh S, Thimmaiah KN et-al .Targeting calmodulin in reversing multi drug resistance in cancer cells. Mini Rev Med Chem.2006;6(12):1383-9
[11]Rosenwald A.Multi-drug resistance in B-cell chronic lymphocytic leukemia (B-CLL):A feature of B-CLL sub-sets with poor prognosis genetic alterations?Leuk Lymphoma.2006;47(11):2263-4
[12]Kang HC,Kim IJ,Park HW et-al.Regulation of MDK expression in human cancer cells modulates sensitivities to various anticancer drugs:MDK overexpression confers to a multi-drug resistance.Cancer Lett.2007;247(1):40-7
[13]陈杰,钱桂生,黄桂君 等.肺癌多药耐药性机制研究进展.中国癌症杂志,2002,12(3):273-276.
[14]Lee JY,Tanabe S,Shimohira H et-al.Expression of cyclooxygenase-2,P-glycoprotein and multi-drug resistance-associated protein in canine transitional cell carcinoma.Res Vet Sci.2007;83(2):210-6
[15]Okada T,Tanaka K,Nakatani F et-al.Involvement of P-glycoprotein and MRP1in resistance to cyclic tetrapeptide subfamily of histone deacetylase inhibitors in the drug-resistant osteosarcoma and Ewing's sarcoma cells.Int J Cancer.2006Jan 1;118(1):90-7
[16]Matsunaga S,Asano T,Tsutsuda-Asano A et-al.Indomethacin overcomes doxorubicin resistance with inhibiting multi-drug resistance protein 1(MRP1).Cancer Chemother Pharmacol.2006;58(3):348-53
[17]Shen H,Paul S,Breuninger L M et-al.Cellular and in Vitro Transport of Glutathione Conjugates by MRP.Biochem.1996;35:5719-5725.
[18]Bouhamyia L,Chantot-Bastaraud S,Zaidi S et-al.Immunolocalization and cell expression of lung resistance-related protein(LRP) in normal and tumoral human respiratory cells.J Histochem Cytochem.2007;55(8):773-82
[19]Scheffer GL,Reurs AW,Jutten B et-al.Selection and characterisation of a phage-displayed human antibody(Fab) reactive to the lung resistance -related major vault protein.Br J Cancer.2002;86(6):954-62
[20]Matsson P,Englund G,Ahlin G et-al.A global drug inhibition pattern for the human ATP-binding cassette transporter breast cancer resistance protein (ABCG2).J Pharmacol Exp Ther.2007;323(1):19-30
[21]Glavinas H,Kis E,P(?)l A et-al.ABCG2(breast cancer resistance protein/mitoxantrone resistance-associated protein) ATPase assay:a useful tool to detect drug-transporter interactions.Drug Metab Dispos.2007;35(9):1533-42
[22]Yeboah D,Kalabis GM,Sun M et-al.Expression and localisation of breast cancer resistance protein(BCRP) in human fetal membranes and decidua and the influence of labour at term.Reprod Fertil Dev.2008;20(2):328-34
[23]Miyamoto KI,Koga-Takeda K,Koga K et-al.Saturable function of P-glycoprotein as a drug-efflux pump in multidrug-resistant tumour cells.J Pharm Pharmacol.1996 May;48(5):522-5
[24]Kage K,Tsukahara S,Sugiyama T et-al.Dominant-negative inhibition of breast cancer resistance protein as drug efflux pump through the inhibition of S-S dependent homodimerization.Int J Cancer.2002;97(5):626-30
[25]O'Connor R.The pharmacology of cancer resistance.Anticancer Res.2007;27(3A):1267-72
[26]Kenneth DT.Glutathione-associated enzymes in anti-cancer drug resisitance[J].Cancer Res,1994,54(16):4313-4320.
[27]Wang AL,Tew KD.Increased glutathione-S-trans-ferase activity in a cell line with acquired resistance tonitrogen mustards[J].Cancer Treat Rep,1985,69(5):677-682.
[28]Nishimura T,Newkirk K,Sessions RB,et al.Immunohisto—chemical staining for glutathione S-transferase predicts responsetO platinum-based chemotherapy in head and neck cancer[J].ClinCancer Res,1996,2(11):1859-1865.
[29]Nakagawa K,Saijo N,Tsuchida S,et al.Glutathione-S-transferase pi as a determinant of drug resistance intransfectant cell lines[J].J Biol Chem,1990,265(8):4296-4301.
[30]钱军,秦叔逵.抗癌新药——榄香烯的药理及临床.中国肿瘤临床,1996年,23(6):453-455.
[31]周洪语,侯菊生,罗其中.榄香烯抗肿瘤作用机制的研究进展.中国肿瘤临床,2000,27(5):392-394.
[32]范钰 林庚金 钱立平 等.榄香烯乳对胃癌SGCO7901细胞株端粒酶活性及细胞凋亡的影响.复旦学报(医学科学版).2001;28(1):5-8
[33]鲍依稀,罗春丽,汤为学 等.榄香烯对人肺腺癌细胞A549作用机理的初步研究.中国肿瘤临床.2002年;29(5):338-341
[34]胡军 金伟 杨佩满.β-榄香烯逆转人乳腺癌MCF27/ADM细胞对阿霉素耐药性的研究.中华肿瘤杂志.2004年,26(5):268-270
[35]胡守友,陈龙邦,王靖华 等.β-榄香烯对小鼠黑色素瘤细胞的诱导分化作用.南京大学学报(自然科学版),1998年;34(5):550-553
[36]陈龙邦,臧静,王靖华.β-榄香烯对荷瘤小鼠免疫功能的影响.肿瘤防治研 究.1998年,25(1)54-56.
[37]张学军,杜权,丁秀萍等.榄香烯乳对r-DNA转录活性的影响及意义.中国肿瘤临床.1999年;26(6):443-444
[38]郦志军,周君富,陈周苗 等.榄香烯对肺癌患者血液SOD活性和LPO浓度的影响研究.中国肿瘤临床1998年;25(11):852-853.
[39]陶磊,周梁,郑璐滢 等.榄香烯对真核细胞翻译起始因子家族表达和血管生成的抑制作用.中华耳鼻咽喉头颈外科杂志.2005年;40(11):840-845.
[40]胡军,金伟,杨佩满.β-榄香烯逆转人乳腺癌MCF27/ADM细胞对阿霉素耐药性的研究.中华肿瘤杂志,2004,26(5):268-270.
[41]王利,魏品康,秦志丰,等.榄香烯对耐药胃癌细胞的逆转实验研究.成都中医药大学学报,2005,28(2):51-53.
[42]王宝成,郭军,狄剑时,等.榄香烯乳剂与肿瘤多药耐药的基础研究.中国肿瘤临床,1996,23(2):143-146.
[43]曾恒,杨彩虹,陈安民.阿霉素冲击诱导人骨肉瘤多药耐药细胞模型.中国矫形外科杂志.2003年(11)5:318-320
[1]Kenneth DT.Glutathione-associated enzymes in anti-cancer drug resisitance[J].Cancer Res,1994,54(16):4313-4320.
[2]Wang AL,Tew KD.Increased glutathione-S-trans-ferase activity in a cell line with acquired resistance tonitrogen mustards[J].Cancer Treat Rep, 1985,69(5):677-682.
[3] TEWKD, GATEL. Glutathione S-transferases as emergingtherapeu-tic targets[J].Expert Opin TherTargets,2001,5(4) :477-489.
[4] Sheehan D,Meade G,Foley VM,et al.Structure,function and evolution of glutathione transferases:im-plications for classification of non-mammalian mem-bers of an ancient enzyme superfamily[J].Biochem J,2001,360(1):1-16.
[5] Nishimura T, Newkirk K, Sessions RB, et al. Immunohisto—chemical staining for glutathione S-transferase predicts responsetO platinum-based chemotherapy in head and neck cancer[J]. ClinCancer Res, 1996, 2(11): 1859—1865.
[6] Nakagawa K,Saijo N,Tsuchida S,et al.Glutathione-S-transferase pi as a determinant of drug resistance intransfectant cell lines[J].J Biol Chem,1990,265(8):4296-4301.
[7] Oakleyl AJ,Lo Bello M,Nuccetelli M,et al.The lig-andin (non-substrate) binding site of human pi classglutathione transferase is located in the electrophilebinding site (H-site) [J].J Mol Biol,1999,291(4):913-926.
[8] Reinemer P, Dirr HW, Ladenstein R,et al. Thethree-dimensional structure of class pi glutathione-S-transferase in complex with glutathione sulfonate at2.3 A resolution[J] .EMBO J,1991,10(10): 1997-2005.
[9] Reinemer P,Dirr HW,Ladenstein R,et al.Three-dimensional structure of class pi glutathione-S-transferase from human placenta in complex with S-hexylglutathione at 2.8 Aresolution[J].J Mol Biol,1992,227(4):214-226.
[10] Luisa R,Robertaf C.Monobromobimane occupies adistinct xenobiotic substrate site in glutathione-Stransferaseπ[J].Protein Science,2003,12(11):2575-2587.
[11] Prade L,Huber R,Manoharan TH.Structures of classpi glutathione-S-transferase from human placenta incomplex with substrate,transition-state analogue and inhibitor[J].Structure,1997,5(10):1287-1295.
[12] Morgan AS,Ciaccio PJ,Tew KD,et al.Isozyme-specific glutathione-S-transferase inhibitors potentiatedrug sensitivity in cultured human cell lines[J],Cancer Chemother Pharmacol,1996,37(5):363-370.
[13] AHOKAS JT, NICHOLLS FA, RAVENSCROFTP J,et al.Inhibi-tion of purified rat liver glutathione S-transferase isozymes by diureticdrugs[J].Biochem Pharmacol,1985,34(12):2157-2161.
[14]PLOEMEN JH,VANOMMEN B,VAN BLADERENP J.Inhibitionof rat and human glutathione S-transferase iso-enzymes by ethacrynicacid and its glutathione conjugate[J].BiochemPharmacol,1990,40(1):1631-1635.
[15]CAFFREY P B,ZHUM,ZHANG Y,et al.Rapid development ofglutathione S-transferase dependent drug resistancein vitroand its prevention by ethacrynic acid[J].Cancer Lett,1999,136(1):47-52.
[16]ZHAO G,YUT,WANG R,et al.Synthesis and structure-activity relationship of ethacrynic acid analogues on glutathione-S-transferase Pl-1 activity inhibition[J].BioorgMed Chem,2005,13(12):4056-4062.
[17]FLATGAARD J E,BAUERKE,KAUVARLM.Isozyme specificity of novel glutathione-S-tranferase inhibitors[J].Cancer Chemother Pharmacol,1993,33(1):63-70.
[18]BURG D,FILIPPOV D V,HERMANNS R,et al.Peptidomimetic glutathione analogues as novelγGT stable GST inhibitors[J].Bioorg Med Chem,2002,10(1):195-205.
[19]MORGAN A S,CIACCIO P J,TEW K D,et al.Isozyme-specific glutathione S-transferase inhibitors potentiate drug sensitivity in cul-tured human tumor cell lines[J].Cancer Chemother Pharmacol,1996,37(4):363-370.
[20]O'BRIEN M L,VULEVIC B,FREER S,et al.Glutathione pep-tidomimetic drugmodulator ofmultidrug resistance-associated protein 1[J].JPharmacol ExpTher,1999,291(3):1348-1355.
[21]GATE L,LUNK A,TEW K D.Resistance to phorbol 12-myristate 13-acetate-induced cell growth arrest in an HL60 cell line chronically exposed to a glutathioneS-transferase p inhibitor[J].BiochemPhar-macol,2003,65(10):1611-1622.
[22]NAKAJIMAT,TAKAYAMAT,MIYANISHI K,et al.Reversal of multiple drug resistance in cholangiocarcinoma by the glutathione S-transferase-π-specific inhibitor O1-hexadecyl-γ-glutamyl-S-benzylcys-teinyl-D-phenylglycine ethyleste[J].J Pharmacol Exp Ther,2003,306(3):861-869.
[23]VAN ZZNDEN J J,BENHAMMAN O,VAN IERSEL M L,et al.Inhibition of human glutathione S-transferase p1-1 by the flavonoid quercetin[J].Chem Biol Interact,2003,145(2):139-148.
[24]VAN ZDNDEN J J,GERAETS L,WORTELBOER HM,et al.Struc-tural requirements for the flavonoid-mediated modulation of glutathione S-transferase P1-1 andGS-Xpump activity inMCF7 breast cancer cells[J].BiochemPharmacol,2004,67(8):1607-1617.
[25]LYON R P,HILL J J,ATK1NS W M.Novel class of bivalent glu-tathione S-transferase inhibitors[J].Biochemistry,2003,42(35):10418-10428.
[26]MUKANGANYAMA S,WIDERSTEN M,NAIK Y S,et al.Inhibi-tion of glutathione S-transferases by antimalarial drugs possible impli-cations for circumventing anticancer drugresistance[J].IntJCancer,2002,97(5):700-705.
[27]VANHAAFTEN R I,EVELO C T,HAENEN G R,et al.α-Toco-pherol inhibits human glutathione S-transferase p[J].Biochem Bio-phys Res Commun,2001,280(3):631-633.
[28]Lyttle MH,Satyam A,Hocker MD,et al.Glutathione-S-transferase activaties novel alkylating agents[J].J Med Chem,1994,37(10):1501-1507.
[29]Rosario LA,Brien ML,Henderson CJ,et al.Cellular response to a glutathione-S-transferaseπactivated prodrug[J].Mol Pharmacol,2000,58(1):167-174.
[30]Hongxie S,Alexandra LS.Efficacy of a glutathione-S-transferaseπactivated prodrug in platinum-resistantovarian cancer cells[J].Mol Cancer Ther,2002,1(12):1089-1095.
[31]Saavedra JE,Shami PJ,Wang LY,et al.Esterase-sensi-tive nitric oxide donors of the diazeniumdiolate familyinvitroantileukemic activity[J].J Med Chem,2000,43(2):261-269.
[32]Shami PJ,Joseph ES,Lai YW,et al.JS-K,a glu-tathione/glutathione-S-transferase -activated nitric ox-ide donor of the diazeniumdiolate class with potent antineoplastic activity[J].Mol Cancer Ther,2003,2(4):409-417.
[33]Victoria JF,Danyelle MT,Joseph ES,et al.Tumor cell responses to a novel glutathione-S-transferase ac-tivated nitric oxide-releasing prodrug[J].Mol Phar- macol,2004,65(5):1070-1078.
[34]Takaharu N,Tetsuji T,Koji M,et al.Reversal of multi-pie drug resistance in cholangiocarcinoma by the glu-tathione-S-transferase-nspecific inhibitor Ol-hexadecyl-γ-glutamyl-S-benzylcysteinyl-D-phenylglycine ethylester[J].J Pharm Exp Ther,2003,306(11):861-869.
[35]Duvoix A,Morceau F,Delhalle S,et al.Induction of apoptosis by curcumin:mediation by glutathione S-transferase P1-1 inhibition[J].Biochem Pharmacol,2003,66(11):1475-1483.
[36]Goto S,Kamada K,Soh Y,et al.Signifieance of nu-clear glutathione-S-transferase pi in resistance to anti- cancer drugs[J].Jpn J Cancer Res,2002,93(10):1047-1056.
[37]Enokida H,Shiina H,Urakami S,et al.Muhigene methylation analysis for detection and staging of prostate cancer[J].Clin Canc—er Res,2005,11(18):6582-6588.
[38]Kim J,Lee HS,Bae SI,et al.Silencing and CpG island methyla—tion of GSTP1 is rare in ordinary gastric carcinomas but corn mon in Epstein-Barr virus—associated gastric carcinomas[J].Anticancer Res,2005,25(6B):4013-4019.
[39]张强,陈梓甫,陈文榜,等.谷胱甘肽S转移酶 在肾细胞癌的表达及临床意义[J].福建医科大学学报,1999,33(4):372-375.
[40]Cullen KJ,Newkirk KA,Schumaker LM,et al.Glutathione Stransferase pi amplification is associated with cisplatin resistance in head and neck squamous cell carcinoma cell lines and primary tumors[J].Cancer Res,2003,63(23):8097-8102.
[41]Procopio A,Alcaro S,Cundari S,et al.Molecular modeling,synthesis,and preliminary biological evaluation of glutathione-Stransferase inhibitors as potential therapeutic agents[J].J Med Chem,2005,48(19):6084-6089.
[2] Barok M, Isola J, P(?)lyi-Krekk Z et-al.Trastuzumab causes antibody -dependent cellular cytotoxicity-mediated growth inhibition of submacroscopic JIMT-1 breast cancer xenografts despite intrinsic drug resistance. Mol Cancer Ther.2007 Jul;6(7):2065-72.
[3] De Anta JM, Perez-Castro AJ, Freire R et-al.The DNA damage checkpoint is activated during residual tumour cell survival to methotrexate treatment as an initial step of acquired drug resistance. Anticancer Drugs. 2006;17(10):1171-7
[4] Garc(?)a-Mart(?)n E, Pizarro RM, Martinez C et-al.Acquired resistance to the anticancer drug paclitaxel is associated with induction of cytochrome P450 2C8.Pharmacogenomics. 2006;7(4):575-85.
[5] Brozovic A, Fritz G, Christmann M et-al. Long-term activation of SAPK/JNK,p38 kinase and fas-L expression by cisplatin is attenuated in human carcinoma cells that acquired drug resistance. Int J Cancer. 2004; 112(6):974-85
[6] Friedrich K, Wieder T, Von Haefen C et-al. Overexpression of caspase-3 restores sensitivity for drug-induced apoptosis in breast cancer cell lines with acquired drug resistance. Oncogene. 2001;20(22):2749-60.
[7] Tian K, Jurukovski V, Wang XP et-al.Epigenetic regulation of WTH3 in primary and cultured drug-resistant breast cancer cells. Cancer Res.2005;65(21):10024-31
[8] Rellecke P, Kuchelmeister K, Schachenmayr W et-al.Mismatch repair protein hMSH2 in primary drug resistance in in vitro human malignant gliomas.J Neurosurg. 2004;101(4):653-8.
[9] Garcia R, Franklin RA, McCubrey JA et-al. EGF induces cell motility and multi-drug resistance gene expression in breast cancer cells.Cell Cycle.2006;5(23):2820-6.
[10] Mayur YC, Jagadeesh S, Thimmaiah KN et-al .Targeting calmodulin in reversing multi drug resistance in cancer cells. Mini Rev Med Chem.2006;6(12):1383-9
[11]Rosenwald A.Multi-drug resistance in B-cell chronic lymphocytic leukemia (B-CLL):A feature of B-CLL sub-sets with poor prognosis genetic alterations?Leuk Lymphoma.2006;47(11):2263-4
[12]Kang HC,Kim IJ,Park HW et-al.Regulation of MDK expression in human cancer cells modulates sensitivities to various anticancer drugs:MDK overexpression confers to a multi-drug resistance.Cancer Lett.2007;247(1):40-7
[13]陈杰,钱桂生,黄桂君 等.肺癌多药耐药性机制研究进展.中国癌症杂志,2002,12(3):273-276.
[14]Lee JY,Tanabe S,Shimohira H et-al.Expression of cyclooxygenase-2,P-glycoprotein and multi-drug resistance-associated protein in canine transitional cell carcinoma.Res Vet Sci.2007;83(2):210-6
[15]Okada T,Tanaka K,Nakatani F et-al.Involvement of P-glycoprotein and MRP1in resistance to cyclic tetrapeptide subfamily of histone deacetylase inhibitors in the drug-resistant osteosarcoma and Ewing's sarcoma cells.Int J Cancer.2006Jan 1;118(1):90-7
[16]Matsunaga S,Asano T,Tsutsuda-Asano A et-al.Indomethacin overcomes doxorubicin resistance with inhibiting multi-drug resistance protein 1(MRP1).Cancer Chemother Pharmacol.2006;58(3):348-53
[17]Shen H,Paul S,Breuninger L M et-al.Cellular and in Vitro Transport of Glutathione Conjugates by MRP.Biochem.1996;35:5719-5725.
[18]Bouhamyia L,Chantot-Bastaraud S,Zaidi S et-al.Immunolocalization and cell expression of lung resistance-related protein(LRP) in normal and tumoral human respiratory cells.J Histochem Cytochem.2007;55(8):773-82
[19]Scheffer GL,Reurs AW,Jutten B et-al.Selection and characterisation of a phage-displayed human antibody(Fab) reactive to the lung resistance -related major vault protein.Br J Cancer.2002;86(6):954-62
[20]Matsson P,Englund G,Ahlin G et-al.A global drug inhibition pattern for the human ATP-binding cassette transporter breast cancer resistance protein (ABCG2).J Pharmacol Exp Ther.2007;323(1):19-30
[21]Glavinas H,Kis E,P(?)l A et-al.ABCG2(breast cancer resistance protein/mitoxantrone resistance-associated protein) ATPase assay:a useful tool to detect drug-transporter interactions.Drug Metab Dispos.2007;35(9):1533-42
[22]Yeboah D,Kalabis GM,Sun M et-al.Expression and localisation of breast cancer resistance protein(BCRP) in human fetal membranes and decidua and the influence of labour at term.Reprod Fertil Dev.2008;20(2):328-34
[23]Miyamoto KI,Koga-Takeda K,Koga K et-al.Saturable function of P-glycoprotein as a drug-efflux pump in multidrug-resistant tumour cells.J Pharm Pharmacol.1996 May;48(5):522-5
[24]Kage K,Tsukahara S,Sugiyama T et-al.Dominant-negative inhibition of breast cancer resistance protein as drug efflux pump through the inhibition of S-S dependent homodimerization.Int J Cancer.2002;97(5):626-30
[25]O'Connor R.The pharmacology of cancer resistance.Anticancer Res.2007;27(3A):1267-72
[26]Kenneth DT.Glutathione-associated enzymes in anti-cancer drug resisitance[J].Cancer Res,1994,54(16):4313-4320.
[27]Wang AL,Tew KD.Increased glutathione-S-trans-ferase activity in a cell line with acquired resistance tonitrogen mustards[J].Cancer Treat Rep,1985,69(5):677-682.
[28]Nishimura T,Newkirk K,Sessions RB,et al.Immunohisto—chemical staining for glutathione S-transferase predicts responsetO platinum-based chemotherapy in head and neck cancer[J].ClinCancer Res,1996,2(11):1859-1865.
[29]Nakagawa K,Saijo N,Tsuchida S,et al.Glutathione-S-transferase pi as a determinant of drug resistance intransfectant cell lines[J].J Biol Chem,1990,265(8):4296-4301.
[30]钱军,秦叔逵.抗癌新药——榄香烯的药理及临床.中国肿瘤临床,1996年,23(6):453-455.
[31]周洪语,侯菊生,罗其中.榄香烯抗肿瘤作用机制的研究进展.中国肿瘤临床,2000,27(5):392-394.
[32]范钰 林庚金 钱立平 等.榄香烯乳对胃癌SGCO7901细胞株端粒酶活性及细胞凋亡的影响.复旦学报(医学科学版).2001;28(1):5-8
[33]鲍依稀,罗春丽,汤为学 等.榄香烯对人肺腺癌细胞A549作用机理的初步研究.中国肿瘤临床.2002年;29(5):338-341
[34]胡军 金伟 杨佩满.β-榄香烯逆转人乳腺癌MCF27/ADM细胞对阿霉素耐药性的研究.中华肿瘤杂志.2004年,26(5):268-270
[35]胡守友,陈龙邦,王靖华 等.β-榄香烯对小鼠黑色素瘤细胞的诱导分化作用.南京大学学报(自然科学版),1998年;34(5):550-553
[36]陈龙邦,臧静,王靖华.β-榄香烯对荷瘤小鼠免疫功能的影响.肿瘤防治研 究.1998年,25(1)54-56.
[37]张学军,杜权,丁秀萍等.榄香烯乳对r-DNA转录活性的影响及意义.中国肿瘤临床.1999年;26(6):443-444
[38]郦志军,周君富,陈周苗 等.榄香烯对肺癌患者血液SOD活性和LPO浓度的影响研究.中国肿瘤临床1998年;25(11):852-853.
[39]陶磊,周梁,郑璐滢 等.榄香烯对真核细胞翻译起始因子家族表达和血管生成的抑制作用.中华耳鼻咽喉头颈外科杂志.2005年;40(11):840-845.
[40]胡军,金伟,杨佩满.β-榄香烯逆转人乳腺癌MCF27/ADM细胞对阿霉素耐药性的研究.中华肿瘤杂志,2004,26(5):268-270.
[41]王利,魏品康,秦志丰,等.榄香烯对耐药胃癌细胞的逆转实验研究.成都中医药大学学报,2005,28(2):51-53.
[42]王宝成,郭军,狄剑时,等.榄香烯乳剂与肿瘤多药耐药的基础研究.中国肿瘤临床,1996,23(2):143-146.
[43]曾恒,杨彩虹,陈安民.阿霉素冲击诱导人骨肉瘤多药耐药细胞模型.中国矫形外科杂志.2003年(11)5:318-320
[1]Kenneth DT.Glutathione-associated enzymes in anti-cancer drug resisitance[J].Cancer Res,1994,54(16):4313-4320.
[2]Wang AL,Tew KD.Increased glutathione-S-trans-ferase activity in a cell line with acquired resistance tonitrogen mustards[J].Cancer Treat Rep, 1985,69(5):677-682.
[3] TEWKD, GATEL. Glutathione S-transferases as emergingtherapeu-tic targets[J].Expert Opin TherTargets,2001,5(4) :477-489.
[4] Sheehan D,Meade G,Foley VM,et al.Structure,function and evolution of glutathione transferases:im-plications for classification of non-mammalian mem-bers of an ancient enzyme superfamily[J].Biochem J,2001,360(1):1-16.
[5] Nishimura T, Newkirk K, Sessions RB, et al. Immunohisto—chemical staining for glutathione S-transferase predicts responsetO platinum-based chemotherapy in head and neck cancer[J]. ClinCancer Res, 1996, 2(11): 1859—1865.
[6] Nakagawa K,Saijo N,Tsuchida S,et al.Glutathione-S-transferase pi as a determinant of drug resistance intransfectant cell lines[J].J Biol Chem,1990,265(8):4296-4301.
[7] Oakleyl AJ,Lo Bello M,Nuccetelli M,et al.The lig-andin (non-substrate) binding site of human pi classglutathione transferase is located in the electrophilebinding site (H-site) [J].J Mol Biol,1999,291(4):913-926.
[8] Reinemer P, Dirr HW, Ladenstein R,et al. Thethree-dimensional structure of class pi glutathione-S-transferase in complex with glutathione sulfonate at2.3 A resolution[J] .EMBO J,1991,10(10): 1997-2005.
[9] Reinemer P,Dirr HW,Ladenstein R,et al.Three-dimensional structure of class pi glutathione-S-transferase from human placenta in complex with S-hexylglutathione at 2.8 Aresolution[J].J Mol Biol,1992,227(4):214-226.
[10] Luisa R,Robertaf C.Monobromobimane occupies adistinct xenobiotic substrate site in glutathione-Stransferaseπ[J].Protein Science,2003,12(11):2575-2587.
[11] Prade L,Huber R,Manoharan TH.Structures of classpi glutathione-S-transferase from human placenta incomplex with substrate,transition-state analogue and inhibitor[J].Structure,1997,5(10):1287-1295.
[12] Morgan AS,Ciaccio PJ,Tew KD,et al.Isozyme-specific glutathione-S-transferase inhibitors potentiatedrug sensitivity in cultured human cell lines[J],Cancer Chemother Pharmacol,1996,37(5):363-370.
[13] AHOKAS JT, NICHOLLS FA, RAVENSCROFTP J,et al.Inhibi-tion of purified rat liver glutathione S-transferase isozymes by diureticdrugs[J].Biochem Pharmacol,1985,34(12):2157-2161.
[14]PLOEMEN JH,VANOMMEN B,VAN BLADERENP J.Inhibitionof rat and human glutathione S-transferase iso-enzymes by ethacrynicacid and its glutathione conjugate[J].BiochemPharmacol,1990,40(1):1631-1635.
[15]CAFFREY P B,ZHUM,ZHANG Y,et al.Rapid development ofglutathione S-transferase dependent drug resistancein vitroand its prevention by ethacrynic acid[J].Cancer Lett,1999,136(1):47-52.
[16]ZHAO G,YUT,WANG R,et al.Synthesis and structure-activity relationship of ethacrynic acid analogues on glutathione-S-transferase Pl-1 activity inhibition[J].BioorgMed Chem,2005,13(12):4056-4062.
[17]FLATGAARD J E,BAUERKE,KAUVARLM.Isozyme specificity of novel glutathione-S-tranferase inhibitors[J].Cancer Chemother Pharmacol,1993,33(1):63-70.
[18]BURG D,FILIPPOV D V,HERMANNS R,et al.Peptidomimetic glutathione analogues as novelγGT stable GST inhibitors[J].Bioorg Med Chem,2002,10(1):195-205.
[19]MORGAN A S,CIACCIO P J,TEW K D,et al.Isozyme-specific glutathione S-transferase inhibitors potentiate drug sensitivity in cul-tured human tumor cell lines[J].Cancer Chemother Pharmacol,1996,37(4):363-370.
[20]O'BRIEN M L,VULEVIC B,FREER S,et al.Glutathione pep-tidomimetic drugmodulator ofmultidrug resistance-associated protein 1[J].JPharmacol ExpTher,1999,291(3):1348-1355.
[21]GATE L,LUNK A,TEW K D.Resistance to phorbol 12-myristate 13-acetate-induced cell growth arrest in an HL60 cell line chronically exposed to a glutathioneS-transferase p inhibitor[J].BiochemPhar-macol,2003,65(10):1611-1622.
[22]NAKAJIMAT,TAKAYAMAT,MIYANISHI K,et al.Reversal of multiple drug resistance in cholangiocarcinoma by the glutathione S-transferase-π-specific inhibitor O1-hexadecyl-γ-glutamyl-S-benzylcys-teinyl-D-phenylglycine ethyleste[J].J Pharmacol Exp Ther,2003,306(3):861-869.
[23]VAN ZZNDEN J J,BENHAMMAN O,VAN IERSEL M L,et al.Inhibition of human glutathione S-transferase p1-1 by the flavonoid quercetin[J].Chem Biol Interact,2003,145(2):139-148.
[24]VAN ZDNDEN J J,GERAETS L,WORTELBOER HM,et al.Struc-tural requirements for the flavonoid-mediated modulation of glutathione S-transferase P1-1 andGS-Xpump activity inMCF7 breast cancer cells[J].BiochemPharmacol,2004,67(8):1607-1617.
[25]LYON R P,HILL J J,ATK1NS W M.Novel class of bivalent glu-tathione S-transferase inhibitors[J].Biochemistry,2003,42(35):10418-10428.
[26]MUKANGANYAMA S,WIDERSTEN M,NAIK Y S,et al.Inhibi-tion of glutathione S-transferases by antimalarial drugs possible impli-cations for circumventing anticancer drugresistance[J].IntJCancer,2002,97(5):700-705.
[27]VANHAAFTEN R I,EVELO C T,HAENEN G R,et al.α-Toco-pherol inhibits human glutathione S-transferase p[J].Biochem Bio-phys Res Commun,2001,280(3):631-633.
[28]Lyttle MH,Satyam A,Hocker MD,et al.Glutathione-S-transferase activaties novel alkylating agents[J].J Med Chem,1994,37(10):1501-1507.
[29]Rosario LA,Brien ML,Henderson CJ,et al.Cellular response to a glutathione-S-transferaseπactivated prodrug[J].Mol Pharmacol,2000,58(1):167-174.
[30]Hongxie S,Alexandra LS.Efficacy of a glutathione-S-transferaseπactivated prodrug in platinum-resistantovarian cancer cells[J].Mol Cancer Ther,2002,1(12):1089-1095.
[31]Saavedra JE,Shami PJ,Wang LY,et al.Esterase-sensi-tive nitric oxide donors of the diazeniumdiolate familyinvitroantileukemic activity[J].J Med Chem,2000,43(2):261-269.
[32]Shami PJ,Joseph ES,Lai YW,et al.JS-K,a glu-tathione/glutathione-S-transferase -activated nitric ox-ide donor of the diazeniumdiolate class with potent antineoplastic activity[J].Mol Cancer Ther,2003,2(4):409-417.
[33]Victoria JF,Danyelle MT,Joseph ES,et al.Tumor cell responses to a novel glutathione-S-transferase ac-tivated nitric oxide-releasing prodrug[J].Mol Phar- macol,2004,65(5):1070-1078.
[34]Takaharu N,Tetsuji T,Koji M,et al.Reversal of multi-pie drug resistance in cholangiocarcinoma by the glu-tathione-S-transferase-nspecific inhibitor Ol-hexadecyl-γ-glutamyl-S-benzylcysteinyl-D-phenylglycine ethylester[J].J Pharm Exp Ther,2003,306(11):861-869.
[35]Duvoix A,Morceau F,Delhalle S,et al.Induction of apoptosis by curcumin:mediation by glutathione S-transferase P1-1 inhibition[J].Biochem Pharmacol,2003,66(11):1475-1483.
[36]Goto S,Kamada K,Soh Y,et al.Signifieance of nu-clear glutathione-S-transferase pi in resistance to anti- cancer drugs[J].Jpn J Cancer Res,2002,93(10):1047-1056.
[37]Enokida H,Shiina H,Urakami S,et al.Muhigene methylation analysis for detection and staging of prostate cancer[J].Clin Canc—er Res,2005,11(18):6582-6588.
[38]Kim J,Lee HS,Bae SI,et al.Silencing and CpG island methyla—tion of GSTP1 is rare in ordinary gastric carcinomas but corn mon in Epstein-Barr virus—associated gastric carcinomas[J].Anticancer Res,2005,25(6B):4013-4019.
[39]张强,陈梓甫,陈文榜,等.谷胱甘肽S转移酶 在肾细胞癌的表达及临床意义[J].福建医科大学学报,1999,33(4):372-375.
[40]Cullen KJ,Newkirk KA,Schumaker LM,et al.Glutathione Stransferase pi amplification is associated with cisplatin resistance in head and neck squamous cell carcinoma cell lines and primary tumors[J].Cancer Res,2003,63(23):8097-8102.
[41]Procopio A,Alcaro S,Cundari S,et al.Molecular modeling,synthesis,and preliminary biological evaluation of glutathione-Stransferase inhibitors as potential therapeutic agents[J].J Med Chem,2005,48(19):6084-6089.