肿瘤多药耐药性及其逆转策略

 肿瘤细胞多药耐药性（multidrug resistance，MDR）的产生是导致肿瘤化疗失败的主要原因之一。MDR是指肿瘤细胞对一种抗肿瘤药物产生耐药性的同时，对结构和作用机制完全不同的其他抗肿瘤药物产生交叉耐药性的现象。^[1]      

肿瘤多药耐药性的研究进展

肿瘤的多药耐药性(multidrug resistance,MDR)是指肿瘤细胞接触一种抗肿瘤药物并产生耐药后,同时对结构和作用机理不同的多种天然来源的抗肿瘤药物具有交叉耐药性。MDR是肿瘤细胞耐药的常见方式,也是肿瘤化疗失败的主要原因。因此,MDR是当前肿瘤化疗中亟待解决的难题。MDR形成的机理十分复杂,肿瘤细胞可以通过不同途径导致MDR的产生。本文介绍MDR的研究进展。     

NF-κB与肿瘤细胞多药耐药性

肿瘤细胞对抗癌药物产生耐药性，尤其是多药耐药性是肿瘤化疗失败的主要原因之一。所谓多药耐药性(muhidrug resistance，MDR)是指肿瘤细胞对一种抗肿瘤药物产生耐药性，同时对结构和作用机理不同的多种天然来源的抗肿瘤药物产生交叉耐药性。目前，研究MDR的机理成为肿瘤研究中的热点。近年来随着研究的深入，发现耐药的发生与肿瘤细胞的凋亡(apoptosis)有关。     

肿瘤的多药耐药性

 耐药性是肿瘤化疗失败的主要原因之一，它可以存在于化疗之前(即内源性耐药)，也可以在化疗中产生(即获得性耐药)^[1].多药耐药性(Multidrug Resistance，简称MDR)是指肿瘤细胞在接触一种抗癌药物后产生了对其它多种结构与功能迥异的天熊杭痛药物的耐药^[2]。     

多药抗药性及其扭转剂的进展

多药抗药性（multidrugresistanceMDR）是指肿瘤细胞对一种抗肿瘤药物产生抗药性的同时，对结构和作用不同的天然肿瘤药物产生交叉抗药性。多药抗药性是肿瘤化疗失败的主要原因，也是肿瘤化疗急需解决的难题。多药抗药性的形成机理复杂。肿瘤细胞可通过不同途径导致MDR的产生。同时，单个MDR细胞可同时存在多种抗药性的机制。任何一种或多种机制联合作用均可导致MDR产生。一、细胞膜蛋白质与MDR在MDR细胞膜上最常见的变化是P-糖蛋白（PermeabilityglycoproteinP－gp）的过度表达，此外尚有其他膜蛋白变化的报道。（一）P-gp与MDR…     

逆转胃肠道肿瘤多药耐药性的研究进展

化疗是治疗人体恶性肿瘤的重要手段之一 ,然而肿瘤细胞耐药性常常限制了疗效的提高。自从发现多药耐药 (ＭＤＲ )现象以来 ,国内外对ＭＤＲ进行了广泛、深入的实验和临床研究 ,证明其机制包括 :多药耐药基因 (ＭＤＲ 1基因 )及其编码的Ｐ -糖蛋白 (Ｐ ｇｐ)过度表达 ;多药耐药相关蛋白 (ＭＲＰ)表达增加 ;谷胱甘肽 (ＧＳＨ )依赖性解毒酶系统活性增加 ;蛋白激酶Ｃ变化 ;ＤＮＡ拓朴异构酶含量减少或性质发生改变等。ＭＤＲ逆转治疗是克服肿瘤耐药、提高化疗效果的 1种重要手段。近年来寻求拮抗或逆转肿瘤细胞ＭＤＲ的研究颇多 ,从报道异搏定…     

caveolae与肿瘤细胞的多药耐药性

 caveolae是特化的细胞膜微结构域，它由其特异性的被覆蛋白caveolin及多种脂类分子和膜蛋白组成，在细胞外分子的内化、信号的跨膜转导和胆固醇的转运过程中起着重要的作用。新近的研究表明，caveolae及其某些组成成分在肿瘤多药耐药细胞中表达上调，并有可能参与了肿瘤细胞多药耐药性的形成。对caveolae与肿瘤细胞多药耐药性的研究进展进行了综述。     

NF-κB与肿瘤细胞多药耐药性

肿瘤细胞对抗癌药物产生耐药性 ,尤其是多药耐药性是肿瘤化疗失败的主要原因之一。所谓多药耐药性 (multidrugresistance ,MDR)是指肿瘤细胞对一种抗肿瘤药物产生耐药性 ,同时对结构和作用机理不同的多种天然来源的抗肿瘤药物产生交叉耐药性。目前 ,研究MDR的机理成为肿瘤研究中的热点[1] 。近年来随着研究的深入 ,发现耐药的发生与肿瘤细胞的凋亡(apoptosis)有关[2 ] 。经过近几年的研究已发现p5 3突变 ,bcl 2基因 ,Fas/FasL ,caspase蛋白酶家族等均与多药耐药性的发生过程有关[2 ] 。核转录因子kappaB(NF κB)是一组重要的转录调节…     

多药耐药性的研究进展

多药耐药性的研究进展上海医科大学肿瘤研究所（２０００３２）韩军综述许良中审阅化学药物治疗作为一种全身性治疗手段，在恶性肿瘤这一全身性疾病的综合治疗中有着很重要的地位和发展前景。耐药性是临床肿瘤化疗失败最常见和最难以克服的问题之一。肿瘤细胞耐药的方式较...     

逆转肿瘤多药耐药性的系列研究

多药耐药性（ｍｕｌｔｉｄｒｕｇ ｒｅｓｉｓｔａｎｃｅ, ＭＤＲ）是指肿瘤细胞接触一种来源于天然的抗肿瘤药物并产生耐药性的同时,对结构和作用机理不同的多种来源于天然的抗肿瘤药物产生交叉耐药性。多药耐药性是肿瘤化疗失败的主要原因。ＭＤＲ产生的原因复杂,但ｍｄｒｌ基因过度表达产生的Ｐ－糖蛋白（Ｐｅｒｍｅａｂｉｌｉｔｙ　 ｇｌｙｃｏｐｒｏｔｅｉｎ, Ｐ－ｇｐ）是 ＭＤＲ产生的最重要和最常见的原因。实际上,Ｐ－ｇｐ是一种能量依赖性药物转出泵,能将化疗药物从癌细胞内泵出细胞外,从而使药物在细胞内积蓄浓度降低,使…     

乳腺癌干细胞多药耐药基因的表达及意义

Objective:To approach the expressions of MDR1 and BCRP in breast cancer stem cells and differentiated cells.Methods:The breast cancer stem calls were separated from human breast cancer primary tissues and MCF-7 by flow cytometry.Then we measured the expressions of MDR1 and BCRP with different subset cells by Realtime-PCR.Results:Contrasted with breast cancer differentiated cells,the expressions of MDR1 and BCRP in breast cancer stem calls were higher (P＜0.01),and the proportion of stem cells rose after chemotherapy (P＜0.01).Conclusion:Contrasted with breast cancer differentiated cells,breast cancer stem cells have stronger ability of clrug-resistanca with higher level of multi-drug resistance genes,and it is one of key points for chemotherapy failure of breast cancer.     

microRNAs与ABC转运蛋白介导的肿瘤多药耐药研究进展

目的回顾ABC转运蛋白与肿瘤耐药的研究现状,探讨miRNA在逆转肿瘤耐药过程中的作用机制。方法应用PubMed和CNKI期刊全文数据库检索系统,检索2010-01-01-2014-05-20的相关文献,以"ABC转运蛋白、miRNA和多药耐药"为关键词。纳入标准:1)ABC转运蛋白的表达水平与肿瘤耐药;2)miRNA对ABC转运蛋白表达水平的调控;3)miRNA对肿瘤细胞药物敏感性的影响,根据纳入标准符合分析的文献34篇。结果大多数癌症患者使用一种化疗药物治疗后,肿瘤细胞可能因为种种原因,不仅对该药产生耐药,而且对多种结构不同和作用机制完全不同的其他药物也产生交叉耐药。研究表明,在多药耐药导致肿瘤化疗失败的众多原因中,ABC转运蛋白过表达是导致肿瘤多药耐药的主要原因之一。在耐药肿瘤细胞当中高表达的ABC转运蛋白主要有乳腺癌耐药蛋白ABCG2,多药耐药相关蛋白ABCC1,P-糖蛋白ABCB1,这些蛋白采用ATP水解的能量将细胞内药物泵出细胞外,从而降低细胞内药物的浓度,使细胞产生耐药性。miRNA能与ABC转运蛋白mRNA的3′UTR结合,使mRNA降解或抑制其翻译,导致目标蛋白的表达受到抑制,从而增加肿瘤细胞的药物敏感性,逆转由ABC转运蛋白过表达引起的肿瘤多药耐药。结论 miRNA可以逆转由ABC转运蛋白家族高表达所引起的肿瘤耐药,这为肿瘤多药耐药的研究提供了新的思路。     

肿瘤多药耐药机制的研究进展

多药耐药 (Multi drugresistance ,MDR)是肿瘤化疗失败的主要原因之一。从药物转运、药物代谢、药物靶、细胞凋亡及凋亡相关基因 4个方面 ,全面复习近年来有关MDR的机制 ,为肿瘤临床化疗及临床MDR研究提供重要的理论依据     

淀粉样变治疗学研究的最新进展

既往10-15年中系统性淀粉样变新治疗的发展取得了实质性进展。这些进展改善了患者的转归,但亦带来新的临床问题。例如,由于淀粉样轻链(或)(AL)淀粉样变对自体造血干细胞移植的作用不太肯定,为此正在开发广泛途径的治疗研究和提供最佳支持治疗。现认为淀粉样前体和成熟原纤维促使淀粉样变相关的器官病变,显示完全清除AL淀粉样变患者促淀粉样(amyloidogenic)轻链生成的重要性,同时治疗前后需作淀粉样前体蛋白和器官病变标记的监测。     

淀粉样变治疗学研究的最新进展

既往10—15年中系统性淀粉样变新治疗的发展取得了实质性进展。这些进展改善了患者的转归，但亦带来新的临床问题。例如，由于淀粉样轻链（K或入）（AL）淀粉样变对自体造血干细胞移植的作用不太肯定，为此正在开发广泛途径的治疗研究和提供最佳支持治疗。现认为淀粉样前体和成熟原纤维促使淀粉样变相关的器官病变，显示完全清除AL淀粉样变患者促淀粉样（amyloidogenic）轻链生成的重要性，同时治疗前后需作淀粉样前体蛋白和器官病变标记的监测。     

树突状细胞肿瘤疫苗研究进展

树突状细胞(DC)是具有最强抗原提呈能力的专职性抗原提呈细胞.近年来,以DC为基础的肿瘤疫苗研究已成为肿瘤免疫治疗的热点之一.本文综述各种DC肿瘤疫苗的制备方法和目前临床应用现状.     

吉西他滨耐药细胞系H460/Gem及其亲代细胞NCI-H460之间的不同

Objective: To discuss the difference between multi-drug resistant cell line H460/Gem and its parental cell NCI-H460 on the basis of establishment of human gemcitabine-resistant cell line H460/Gem so as to elaborate the possible mech-anisms of gemcitabine resistance. Methods: Human gemcitabine-resistant non-small cell lung cancer cell line H460/Gem was established by 2/3 clinical serous peak concentration gemcitabine intermittent selection from its parental cell human large cell lung carcinoma cell line NCI-H460 which was sensitive to gemcitabine. During the course of inducement, we had monitored their morphology, checked their resistance indexes and resistant pedigree by MTT method, gathered their growth curves and calculated their doubling time, examined their DNA contents and cell cycles by FCM; at the same time, we had measured its expressions of P53, EGFR, c-erb-B-2, PTEN, PCNA, c-myc, VEGF, MDR-1, Bcl-2, nm23, MMP-9, TIMP-1, CD44v6 proteins via immunocytochemistry staining, RRM1 and ERCC1 mRNA by real-time fluorescent quantitative-PCR. Results: The resis-tance index of H460/Gem' cells (the deputy of cells in the process of inducement) to gemcitabine was 1.201, and the cell line also exhibited cross-resistance to paclitaxol, fluorouraci, etoposide, cisplatin and oxaliplatin, but kept sensitivity to vinorelbine and taxotere. The doubling time of H460/Gem' cells was longer and figures in G0-G1 phase was decreased than that of NCI-H460 cells. Compared with NCI-H460 cells, H460/Gem' cells had achieved TIMP-1 protein expression emerged, nm23 protein expression enhanced, VEGF and MMP-9 protein expressions reduced, and CD44v6, P53 protein expressions van-ished, but expressions of EGFR, c-erb-B-2, PTEN, PCNA, c-myc, MDR-1, Bcl-2 proteins and RRM1, ERCC1 mRNA changed trivially. The resistance index of H460/Gem cells to gemcitabine was 1.644, and the cell line also exhibited cross-resistance to fluorouraci, cisplatin and oxaliplatin, but kept sensitivity to paclitaxol, vinorelbine, taxotere, and etoposide. The doubling time of H460/Gem cells was longer and figures in G0-G1 phase was decreased than those of NCI-H460 cells. The farther studies indicated that, compared with NCI-H460 cells, the expressions of MDR-1, nm23 and Bcl-2 proteins in H460/Gem cells had been enhanced, c-erb-B-2 protein expression emerged, P53, MMP-9 and VEGR protein expression had been weakened, but the changes of PTEN, PCNA, c-myc, TIMP-1, EGFR, CD44v6 protein, RRM1 mRNA and ERCC1 mRNA expressions were trivial. Furthermore, compared with its parental cells, H460/Gem cells were mixed with giant cells of different sizes that were larger and more irregular. Conclusion: The human gemcitabine-resistant non-small cell lung cancer cell line H460/Gem had achieved multi-drug resistance and great changes of biological characters compared with its parental cells. And these changes possibly participated in the formation of multidrug resistance.     

Detection and characterisation of multi-drug resistance protein 1 (MRP-1) in human mitochondria

Background:

Overexpression of plasma membrane multi-drug resistance protein 1 (MRP-1) can lead to multidrug resistance. In this study, we describe for the first time the expression of mitochondrial MRP-1 in untreated human normal and cancer cells and tissues.

Methods:

MRP-1 expression and subcellular localisation in normal and cancer cells and tissues was examined by differential centrifugation and western blotting, and immunofluorescence microscopy. Viable mitochondria were isolated and MRP-1 efflux activity measured using the calcein-AM functional assay. MRP-1 expression was increased using retroviral infection and specific overexpression confirmed by RNA array. Cell viability was determined by trypan blue exclusion and annexin V-propidium iodide labelling of cells.

Results:

MRP-1 was detected in the mitochondria of cancer and normal cells and tissues. The efflux activity of mitochondrial MRP-1 was more efficient (55–64%) than that of plasma membrane MRP-1 (11–22% P<0.001). Induced MRP-1 expression resulted in a preferential increase in mitochondrial MRP-1, suggesting selective targeting to this organelle. Treatment with a non-lethal concentration of doxorubicin (0.85 n, 8 h) increased mitochondrial and plasma membrane MRP-1, increasing resistance to MRP-1 substrates. For the first time, we have identified MRP-1 with efflux activity in human mitochondria.

Conclusion:

Mitochondrial MRP-1 may be an exciting new therapeutic target where historically MRP-1 inhibitor strategies have limited clinical success.     

多药耐药细胞株A549/Gem及其亲代细胞A549之间的区别研究

Objective: To discuss the difference between multi-drug resistant cell line A549/Gem and its parental cell A549 on the basis of establishment of human gemcitabine-resistant cell line A549/Gem so as to elaborate the possible mechanisms of gemcitabine resistance. Methods: Human gemcitabine-resistant non-small cell lung cancer cell line A549/Gem was estab-lished by the method of repeated clinical serous peak concentration plus gradually increasing concentration of gemcitabine from its parental cell human lung adenocaroinoma cell line A549 which was sensitive to gemcitabine. During the course of inducement, we had monitored their morphology, checked their resistance indexes and resistant pedigree by MTT method, gathered their growth curves and calculated their doubling time, examined their DNA contents and cell cycles by FCM; at the same time, we had measured their expressions of P53, EGFR, Cerb-B-2, PTEN, PCNA, c-myc, VEGF, MDR-1, Bcl-2, nm23, MMP-9, TIMP-1, and CD44v6 proteins via immunocytochemistry staining, RRM1 and ERCC1 mRNA by real-time fluorescent quantitative-PCR. Results: The resistance index of A549/Gem' cells (the deputy of cells in the process of inducement) to gemcitabine was 163.228, and the cell line also exhibited cross-resistance to vinorelbine, taxotere, fluorouraci, etoposide and cisplatin, but kept sensitivity to paclitaxol and oxaliplatin. The doubling time of A549/Gem' was shorter and figures in G0-G1 phases were increased than A549 cells. Compared with A549 cells, A549/Gem' cells achieved EGFR and c-myc proteins expressions, nm23 protein expression enhanced, P53, Cerb-B-2 and Bcl-2 proteins expressions reduced, PTEN ,PCNA and MDR-1 proteins expressions vanished, but those of MMP-9, VEGF, CD44v6 and TIMP-1 proteins changed trivially. Meanwhile, expressions of RRM1 and ERCC1 mRNA were augmented markedly. The resistance index of A549/Gem cells to gemcitabine was 129.783, and the cell line also held cross-resistance to vinorelbine, taxotere, etoposide, cisplatin and sensitivity to paclitaxol. But the resistance to fiuorouracil and sensitivity to oxaliplatin vanished. And the expression of RRM1 and ERCC1 mRNA decreased visibly. The doubling time of A549/Gem cells was longer and figures in G0-G1 phases were decreased than A549/Gem' cells. In A549/Gem cells, expressions of P53, EGFR, PCNA and MDR-1 proteins was same to those of A549/Gem' cells. A549/Gem cells achieved TIMP-1 and PTEN proteins expressions, Cerb-B-2, MMP-9, c-myc and Bcl-2 proteins expressions enhanced, nm23 protein expressions vanished, but the expressions of VEGF and CD44v6 proteins changed trivially. Furthermore, Compared with its parental cell A549, A549/Gem cell was mixed with giant cells of different sizes and was larger and more irregular. Conclusion: The human gemcitabine-resistant non-small cell lung cancer cell line A549/Gem had achieved multi-drug resistance and great changes of biological characters compared with its parental cells A549. And these changes possibly participated in the formation of multidrug resistance.     

人大细胞肺癌多药耐药株H460/cDDP的建立及其生物学特性

目的：建立人大细胞肺癌多药耐药株H460/cDDP．并对其生物学特性进行测定。方法：以人大细胞肺癌细胞株H460为亲本细胞．采用顺铂大剂量问歇诱导法建立多药耐药细胞株H460／cDDP。光镜下观察其形态学变化；MTT法测定药物敏感性；锥虫蓝拒染实验测定细胞生长曲线、倍增时间；进行染色体核型分析；棵鼠成癌实验测定其体内成瘤活性。结果：历时近6个月建成人大细胞肺癌多药耐药株H460／cDDP．经鉴定其对顺铂的耐药指数为10．21，对5-氟尿嘧啶，多柔比星(阿霉素)、依托泊苷(足叶乙甙)、甲氨蝶呤有不同程度的交叉耐药。H460／cDDP较亲本细胞分裂高峰延迟，倍增时间由20．78小时延长至3646小时．细胞形态改变，染色体变化不突出。经过反复冻存．复苏．上述特性保持稳定．并具有体内成瘤活性，成瘤时间较亲本细胞有所延迟。结论：新建大细胞肺癌多药耐药株H460／cDDP呈中等度耐药、多药耐药表型稳定且具有体内成瘤活性，可用于下游实验。