急性白血病患者多药耐药基团mdrl的表达及其临床意义

探讨白血病细胞的抗药性与化疗效果及预后的关系。采用逆转录－聚合酶链反应的方法检测了８５例不同时期急性白血病患者多药耐药基因ｍｄｒｌ ｍＲＮＡ的表达,同时检测了２０例正常人作为对照。     

急性白血病患者多药耐药基因mdrl的表达及其临床意义

目的　探讨白血病细胞的抗药性与化疗效果及预后的关系。方法　采用逆转录 -聚合酶链反应 ( RT- PCR)的方法检测了 85例不同时期急性白血病 ( AL)患者多药耐药基因 mdrl m RNA的表达 ,同时检测了 2 0例正常人作为对照。结果　初治组 mdrl的阳性率为 4 4 .7% ,mdrl 与 mdrl-患者的完全缓解 ( CR)率分别为 2 3.9%和 88.5% ( P<0 .0 0 5) ,复发及难治组患者 mdrl的阳性率明显高于CR组 ( P<0 .0 0 1) ,复发及难治组 mdrl m RNA的表达水平也高于 CR组 ( P<0 .0 0 1)。CR患者 mdrlm RNA的表达水平逐渐升高可导致临床复发。正常人和长期生存患者 mdrl的阳性率很低。另外发现mdrl的表达与法、美、英协作组 ( FAB)分型有关。结论　mdrl的表达与 AL的化疗效果及预后密切相关 ,它是 AL患者一个重要的不利预后因素。     

Expression and clinical significance of multidrug resistance gene and multidrug resistance-associated protein gene in acute leukemia

Multidrugresistance(MDR)remainsamaj0rcauseoffailureinthechemotherapeutictreatment0facuteleukemia(AL).ClassicalMDRphenotypeisduet0overexpressionofmembrane-b0undglycoprotein(Pl7O)encodedbymultidrugresistancegene(mdrl).However,lowP-glycoproteinlevelswerefrequentlyfoundinc1inicallydrug-resistantleukemia,andl0r2indicatedthatoverexpression0fthemdrlgenecann0tbeacc0untedforallcasesofdrugresistanceinleukenda.Recently,multidrugresistance-ass0ciatedprotein(MRP)genewasfoundt0beassociatedwithdrug-res…     

Sorcin基因和mdr1基因的相关性及临床意义

目的　探讨急性髓细胞系白血病 (AML)患者可溶性耐药相关钙结合蛋白基因 (sorcin)和多药耐药基因 (mdr1)的相关性表达及其与临床耐药和疗效的关系。方法　Northernblot检测K5 6 2 /A0 2细胞株sorcin和mdr1基因的表达 ,用半定量逆转录聚合酶链反应 (RT PCR)检测 6 5例AML患者、2 7例非白血病患者和健康人的sorcin基因和mdr1基因表达 ,并分析sorcin基因和mdr1基因表达的相关性及其临床意义。结果　AML患者中 ,sorcin和mdr1基因表达明显高于正常对照组 ,差异有非常显著性 (P <0 .0 0 1) ;在AML和对照组中 ,sorcin和mdr1基因的表达有较好的相关性 ;sorcin和mdr1基因的共表达与AML患者的临床耐药和预后具有较高的符合率。Sorcin+ 和mdr1+ 组患者临床耐药发生率为 92 .9% ,完全缓解 (CR)率为 7.1% ;sorcin-和mdr1-组患者临床耐药发生率为 8.6 % ,CR率为91.4 %。结论　白血病患者的sorcin和mdr1基因共表达与其临床耐药和预后密切相关 ,可作为检测临床耐药和判断预后的有效手段之一。     

急性白血病中MDR1 MRP和bc1-2基因表达及其临床意义

目的:研究多药耐药基因(MDR1)、多药耐药相关蛋白基因(MRP)和调控细胞凋亡有重要作用的基因(bcl-2)在急性白血病(AL)中的表达以及它们与临床耐药之间的关系。方法:采用地高辛掺入的半定量逆转录聚合酶链反应(RT-PCR)及常规RT-PCR方法检测了54例急性白血病患者和10例正常人骨髓单核细胞中基因的表达情况。结果:54例AL中MDR1/MRP/bcl-2三基因表达阳性率为16.7%(9/54),MDR1/MRP/bcl-2三基因表达均阴性,发生频率为46.3%(25/54)。46例资料完整的白血病患者中三基因表达均阴性者80.0%缓解(CR),MDR1/MRP或MDR1/MRP/bcl-2共表达者无1人CR(P<0.01)。单基因分析表明MDR1、MRP、bcl-2基因表达阳性率分别为28.3%、41.3%和47.8%,其中MDR1阳性者CR率7.7%,明显低于MDR1阴性者CR率72.7%(P<0.01);MRP阳性CR率21.1%,明显低于阴性CR率77.8%(P<0.01);bcl-2阳性CR率36.4%,亦低于阴性CR率70.8%(P<0.05)。结论:MDR1/MRP或MDR1/MRP/bcl-2共表达的患者不易获得CR,白血病患者的耐药除了与MDR1高表达密切相关外,还与非P-糖蛋白(P-gp)介导的MRP及bcl-2表达等因素相关。     

Relevance of mdr1 gene expression in acute myeloid leukemia and comparison of different diagnostic methods.

In an attempt to determine the incidence and clinical relevance of mdr1 gene expression in acute myeloid leukemia (AML), we examined 126 specimens obtained from adult patients with de novo AML by slot blot and immunocytochemistry. We found a high incidence of mdr1 gene expression in newly diagnosed patients (27% by immunocytochemistry and 43% by slot blot). No difference was observed between newly diagnosed patients and relapsed patients. However, patients with resistant disease showed statistically higher incidence of mdr1 gene expression compared to the untreated and relapsing patients (60% versus 27% by immunocytochemistry, p 0.005; and 73% versus 45% by slot blot, p less than 0.05). The expression of mdr1 gene correlated significantly with clinical drug resistance: 62% of patients positive for mdr1-mRNA and 68% of patients positive for P-glycoprotein (P-gp) eventually developed resistance to chemotherapy, while this was the case for a lower percentage of patients who did not express mdr1 gene (only 23% by slot blot analysis, p = 0.0052, or 24% by immunocytochemistry, p = 0.0009). A combined parameter, mdr1-mRNA/P-gp, had a very high prognostic value in terms of specificity and sensitivity. All nine patients (100%) who were mdr1-mRNA+/P-gp+ progressed to clinical drug resistance afterward, whereas 11 of 13 (85%) patients who were mdr1-mRNA-1 P-gp- entered complete remission and only two patients later developed drug resistance (p = 0.0005). It could thus be used as a reliable parameter in clinical settings.     

白血病患者WT1基因表达及其临床意义

目的:探讨WT1基因在白血病中表达情况与临床意义。方法:采用筑巢式逆转录聚合酶链反应(NestRT-PCR)检测K562白血病细胞系、78例急性白血病(AL)患者、10例慢性粒细胞白血病(CML)患者和22例正常对照骨髓或外周血细胞的WT1基因表达。结果:60例初治或复发AL中有44例患者高表达WT1基因,阳性率73.3%。在急性非淋巴细胞白血病(ANLL)和急性淋巴细胞白血病(ALL)阳性率分别为70.8%和83.3%,两组无显著性差异。随着WT1基因表达水平的升高,完全缓解(CR)率有明显下降的趋势,CR后WT1基因表达水平明显下降,复发后又显著升高。结论:用NestRT-PCR测定WT1基因的表达可用作检测白血病微小残留病(MRD)的一项指标。     

Overexpression of glucosylceramide synthase in associated with multidrug resistance of leukemia cells

Ceramide, as a second messenger, initiates one of the major signal transduction pathways in tumor apoptosis. Glucosylceramide synthase (GCS) catalyzes glycosylation of ceramide and produces glucosylceramide. Through GCS, ceramide glycosylation allows cellular escape from ceramide-induced programmed cell death. Here we investigated the expression of GCS in human leukemia cells and an association between GCS and multidrug resistance of leukemia cells. Using RT-PCR technique the level of GCS gene was detected in 65 clinical multidrug resistance/non-resistance cases with leukemia, and in K562 and K562/A02 cell lines. AlamarBlue Assay was applied to confirm the multidrug resistant of K562/A02 cells. PPMP, which is a chemical inhibitor for GCS, was used to determine the relationship between GCS and drug-resistance in K562/A02 cells. In addition, multidrug resistance gene (mdr1), Bcl-2 and Bax mRNA was also analyzed by RT-PCR. The expression of GCS and mdr1 mRNA in clinic multidrug resistance samples exhibited significantly increased compared with clinic drug sensitive group (P<0.05). There was the positive correlation both the expression of GCS and mdr1 genes in leukemia samples (P<0.01, gamma=0.7). AlamarBlue Assay showed that the K562/A02 cell line was 115-fold more resistant to adriamycin and 36-fold more resistant to vincristine compared with drug-sensitive K562 cell line. There also was significant expression difference of GCS and mdr1 genes between K562 and K562/A02 cells. Bcl-2 gene exhibited higher expressions whatever in clinic drug-resistance samples or K562/A02 cells, whereas the expressions of Bax gene were higher in drug-sensitive samples and K562 cells. PPMP increased sensitivity to adriamycin toxicity by inhibiting GCS in K562/A02 cells. Therefore, it is suggested that a high level of GCS in leukemia is possible contributed to multidrug resistance of leukemia cells. Abnormally expressions of the genes in associated with cell apoptosis might be one of the main molecular pathology mechanisms of multidrug resistance caused by GCS gene.     

WT1基因在急性白血病中的表达及其临床意义

 【摘要】　目的　探讨急性白血病（AL）患者体内WT1基因的表达情况及其临床意义。方法　采用荧光定量聚合酶链反应对急性髓系、淋巴细胞白血病66例患者WT1基因进行检测,并比较髓系各亚型之间的基因表达量差异,观察基因表达水平与病程及预后之间的相关性,同时分析造血干细胞移植患者的病程与WT1基因表达之间的关系。结果　66例患者中,87.5 %（14／16）的急性淋巴细胞白血病及76.0 %（38／50）的急性髓系白血病患者WT1基因表达阳性;髓系各亚型中,M3的表达水平低于其他各亚型（与M1、M2、M4、M5相比,P值分别为0.040、0.007、0.006、0.010）。WT1基因的表达水平与疾病状态密切相关,完全缓解组表达水平明显低于未缓解组（P＝0.018）及复发组（P＝0.003）,其滴度的再次升高可提前1.5个月预测复发。WT1基因与造血干细胞移植患者的预后相关,WT1基因转阴的患者较持续表达及一度下降后再次上升的患者预后好。结论　WT1基因在AL患者中的表达可代表微小残留病灶,急性髓系白血病各亚型中M3表达水平相对较低,WT1表达水平与临床病程及预后密切相关。     

急性白血病患者MUC1与WT1基因表达的临床意义

目的：探讨MUC1基因及WT1基因在急性白血病的表达及临床意义。方法：采用逆转录-聚合酶链反应（RT-PCR）方法，检测54例初发或复发急性白血病（AL）患者MUC1基因以及WT1基因的表达．观察MUC1基因及WT1基因的表达与急性白血病患者疗效关系。结果：54例初治或复发AL中MUC1基因表达阳性率50.0％．WT1基因表达阳性率74．1％。MUC1基因阴性表达的初治非M3型AL 19例治疗完全缓解率达94．7％，MUC1阳性表达的初治非M3型AL 18例治疗完全缓解率55.6％；两组比较有显著性差异（P〈0.05）。WT1基因表达阳性与否与化疗疗效无关。结论：MUC1和WT1基因在急性白血病中有一定的表达，MUC1基因阳性表达可作为急性白血病患者预后不良的一项分子生物学指标。     

非M3型急性白血病患者中MUC1基因和MDR1基因表达及其与临床疗效的关系

背景与目的:MUC1基因在胃癌、卵巢癌、多发性骨髓瘤、恶性淋巴瘤等肿瘤中有表达,在急性白血病患者中有较高的表达。但MUC1基因和多药耐药基因(MDR1)相互关系以及两者的表达与急性白血病治疗效果的关系尚有待探讨。本研究拟探讨MUC1基因与MDR1基因表达及其与非M3型急性白血病患者治疗效果的关系。方法:应用逆转录鄄聚合酶链反应(RT鄄PCR)法检测34例初治非M3型急性白血病患者MUC1和MDR1的表达,并观察两种基因表达及其与临床疗效的关系。结果:34例初治非M3型急性白血病患者中MUC1基因阳性率为50%,MDR1基因阳性率为29.4%。MUC1基因阳性患者的MDR1阳性率为52.9%,明显高于MUC1阴性者的5.9%(P=0.003)。MUC1基因阴性者完全缓解(CR)率达94.1%,阳性患者CR率52.9%,两组有显著性差异(P<0.05);MDR1基因阴性者CR率为91.7%,明显高于阳性患者的50.0%(P<0.05)。MUC1基因和MDR1基因均阳性者CR率为55.6%,MUC1基因和MDR1基因均阴性者16例,全部获得CR。结论:非M3型急性白血病MUC1基因阳性者MDR1基因表达率较高,MUC1基因及MDR1基因均为阴性者治疗缓解率高。提示联合检测MUC1基因和MDR1基因对判断初治非M3型急性白血病的疗效有良好的预测作用,可作为临床判断疗效的一项有意义的指标。     

抗凋亡基因bcl-xL与白血病细胞耐药的相关性研究

目的 研究ｂｃｌ－ＸＬ基因高表达的白血病细胞耐药形成中的作用,观察急性髓细胞白血病病人的ｂｃｌ－ＸＬ表达与临床疗效间的相关性。方法 采用脂质体法将ｂｃｌ－ＸＬ ｃＤＮＡ转导入ＨＬ－６０细胞;流式细胞仪定量检测凋亡细胞：ＭＴＴ法测定足叶乙甙、柔红霉素、阿霉素对转染细胞的细胞毒性;ＲＴ－ＰＣＲ法检测２０例急性髓细胞白血病病人的ｂｃｌ－ＸＬ表达。结果 ｂｃｌ－ＸＬ高表达可抑制足叶乙甙诱发的凋亡,抑制率为     

Expression of mdr1, mrp and lrp genes in gastric carcinoma and their clinical significance

Objective: To explore the expression of mdr1, multidrug resistance-associated protein (MRP) and lung resistance protein (LRP) genes in human gastric cancer and their clinical significance. Methods: The mdr1 mRNA was assayed by RT-PCR, the MRP and LRP were detected by flow cytometry. Results: The positive rate of mdr1 mRNA was 44.4% (12/27), and the mean MRP and LRP expression were independent upon patient histologic type, nodal involvement, and TNM stage. The mdr1 mRNA expression in patients with serosa invasion was 30.0% (6/20), much lower than that without serosa invasion (85.7%). Conclusion: The multidrug resistance cells are present in primary gastric carcinomas prior to chemotherapy, and analysis of mdr1 gene, MRP, LRP may have guiding significance in the treatment of gastric carcinoma.     

急性白血病耐药蛋白表达与临床疗效及预后的关系

目的:研究急性白血病(AL)耐药蛋白表达与临床疗效及预后的关系。方法:采用免疫组化法对40例初发AL耐药蛋白PGP、MRP1的表达进行检测,并分析其与病人预后的关系。结果:PGP表达阳性的AL中CR率36.36%,PGP表达阴性的AL中CR率92.85%(P<0.01);MRP1表达阳性的AL中CR率44.44%,MRP1表达阴性的AL中CR率为95.45%(P<0.01);PGP、MRP1同时表达阳性,CR率42.85%,全阴性的AL,CR率100%(P<0.01)。结论:PGP和MRP1的表达与白血病疗效及预后之间存在着一定的相关性。     

儿童急性白血病初治患者LRP和MRP的表达及其临床意义

背景与目的:有研究发现肺耐药相关蛋白(lungresistanceprotein,LRP)和多药耐药相关蛋白(multidrugresistance鄄associatedporotein,MRP)的表达与白血病患儿耐药有关,而且患儿耐药不是由于一种耐药机制引起的。本研究旨在观察LRP和MRP在不同类型小儿白血病初治时的表达情况及其与临床疗效的关系。方法:应用逆转录鄄聚合酶链反应法(RT鄄PCR)检测38例不同类型儿童急性白血病[27例急性淋巴细胞白血病(acutelymphoblastic,ALL),11例急性非淋巴细胞白血病(acutenonlymphocyticleukemia,ANLL)]及6名正常儿童LRP、MRP基因的表达,结合患儿化疗后的完全缓解(CR)率分析两种基因表达的意义。结果:38例患儿化疗后CR26例(68.4%)。38例患儿中LRP基因表达阳性11例(28.9%),表达阴性27例(71.1%),6例正常对照儿童LRP基因阴性。LRP基因阳性患儿的CR率低于阴性者(分别为27.3%和85.2%,P<0.05)。MRP阳性者21例,6例正常对照儿童MRP基因阴性。MRP基因阳性者的CR率低于阴性者(分别为47.6%和94.1%,P<0.05)。27例ALL患儿中LRP阳性5例,11例ANLL患儿中LRP阳性6例(分别为18.5%和54.5%,P<0.05)。27例ALL患儿中MRP阳性16例;11例ANLL患儿MRP阳性5例(分别为59.3%和45.5%,P>0.05),ALL和ANLL中MRP的表达无显著性差异。LRP基因阳性患者的MRP阳性率与LRP阴性者MRP阳性率无显著性差异(分别为28.6%和29.4%,P>0.05),说明LRP基因与MRP基因之间无相关性。结论:LRP基因及MRP基因阳性儿童急性白血病者病情重、化疗缓解率低,儿童ANLL相对于ALL化疗缓解率低可能与LRP的表达有关。     

急性白血病GST-π、MRP1表达的临床研究

 目的 检测急性白血病（AL ）患者谷胱甘肽硫转移酶-π（GST-π）、多药耐药相关蛋白1（MRP1）的表达,探讨二者与AL多药耐药性（MDR）及临床预后的关系。方法 采用免疫组织化学SP法检测80例AL患者及30名正常人外周血单个核细胞GST-π、MRP1 表达。结果 GST-π、MRP1在难治组的阳性率均高于初治组和完全缓解组;初治组的阳性率高于对照组。GST-π、MRP1表达阳性组与阴性组之间的完全缓解率差异有统计学意义。结论 GST-π和MRP1的高表达与AL临床耐药有关,GST-π和MRP1表达阳性的AL患者完全缓解率明显降低,GST-π和MRP1表达时白血病患者预后不良。     

Experimental Study on the Mechanism of Reversal of Leukemia Multidrug Resistance by Proteasome Inhibitor Bortezomib

OBJECTIVE In this study, we applied multidrug resistant leukemia cell line expressing mdr1-mRNA to observe changes in mdr1-mRNA, the P-gp, cell cycle and apoptosis before and after bortezomib was used, in order to explore the mechanism of reversal of leukemia multidrug resistance by the proteasome inhibitor bortezomib.METHODS Flow cytometry (FCM) was used to detect the intracellular drug concentration, expression of P-gp, cell apoptosis and cell cycle status of K562/DNR cells before and a er treatment with different concentrations of bortezomib. Fluorescence quantitative PCR was applied to detect the mdr1-mRNA expression in K562/DNR and K562/S cells.RESULTS Bortezomib could increase the intracellular DNR content in K562/DNR cells, but showed no e. ect in K562/S cells.5-100 nmol/L bortezomib could significantly reduce the P-gp/mdr1-mRNA expression in K562/DNR cells in vitro, and showed a dose-dependent effect. There was a statistically significant di. erence (P < 0.05) between di. erent concentration groups and the control group. P-gp/mdr1-mRNA expression was negatively correlated with cell apoptosis (r = -0.912 and P < 0.01). After treatment with different concentrations of bortezomib for 24 h,K562/DNR cells in G2 + M phases were significantly increased,while cells in G0 + G1 phases and S phase were significantly decreased, accompanied by an increased apoptotic rate.CONCLUSION Bortezomib can induce G0 + G1 phase to G2 + M phase, and thereby enhance the chemosensitivity of leukemia, and may also reverse the multidrug resistance in leukemia mediated by P-gp overexpression encoded by mdr1 gene. This confi rms that bortezomib can reverse leukemia multidrug resistance at the levels of nucleic acid and protein molecules.     

Antitumor effects of cytosine deaminase and thymidine kinase fusion suicide gene under the control of mdr1 promoter in mdr1 positive leukemia cells

The multidrug resistance (mdr) mediated by P-glycoprotein (P-gp), the mdr1 gene product, is one of the major obstacles in leukemia treatment. The present study was designed to explore a suicide gene therapy approach targeting mdr1 for reversal of P-gp-mediated mdr in the mdr positive K562/A02 cells. To study targeted killing effects of cytosine deaminase (CD)-thymidine kinase (TK) fusion suicide gene on multi-drug resistant leukemia, the CD-TK fusion suicide gene expression vector driven by mdr1 promoter was constructed and transferred into K562 and K562/A02 cells using lipofectintrade mark 2000. RT-PCR was used to demonstrate that there were CD and TK genes expression in K562/A02 cells, but not in K562 cells. MTT analysis showed that, compared with that in K562/CDTK, the survival rate of K562/A02-CDTK cells decreased and at the same time the apoptotic rate increased after treatment with GCV and 5-FC (P < 0.05). In vivo studies showed that the tumor volume in the prodrug treated K562/A02-CDTK groups was significantly less than that in the NS-control and K562-CDTK groups (P < 0.05). These findings show that the CD and TK fusion suicide gene expression driven by mdr1 promoter is effective in killing multidrug resistant K562/A02 cells.