急性白血病细胞P-gp LRP的表达及其临床意义

目的：研究急性白血痛细胞P-糖蛋白（P—gp）、肺耐药蛋白（LRP）的表达情况，观察其表达率与临床症状及化疗缓解率的关系。方法：利用P—gp、LRP单克隆抗体，采用流式细胞技术分别测定15例正常对照和79例急性白血病（AL）患者P-gp、LRP的表达率，分析两种蛋白表达的临床意义。结果：初治急性淋巴细胞白血病（ALL）组P—gp、LRP的表达率均高于初治急性髓细胞白血病（AML）组（P〈0．01），复发／难治ALL组P—gp的表达率与复发／难治AML组相比无显著性差异（P〉0．05），而LRP的表达率较复发／难治AML组的表达率高（P〈0．01）。复发／难治组P—gp、LRP的表达率均高于相应的初治组（P〈0．05）。急性白血病患者P—gp、LRP的表达之间无相关性（L=0．0746，P〉0．05）。急性白血病细胞P—gp^＋／LRP^＋组缓解率低于P—gp^＋／LRP^-、P—gp^-／LRP^＋组（P〈0．05），并明显低于P—gp^-／LRP^-组（P〈0．01）。结论：复发／难治组P—gp、LRP的表达率高于初治组，而两者的表达率无相关性。P—gp、LRP表达阳性的患者缓解率低，且易出现髓外浸润。同时检测P—gp、LRP的表达较单独检测一种蛋白更具有临床意义。     

凋亡抑制蛋白XIAP在儿童急性白血病中的表达及其临床意义

目的探讨凋亡抑制蛋白XIAP在儿童急性白血病（AL）中的表达及其临床意义。方法54例AL患儿,其中初诊未治26例,治疗后完全缓解（CR）23例,复发5例。应用免疫组织化学方法检测其骨髓细胞XIAP蛋白表达。对照组为10例非恶性血液病且骨髓正常患儿。结果初诊未治AL患儿骨髓中XIAP蛋白表达水平高于缓解期（P〈0．05）和对照组（P〈0．05）,和复发组相比差异无统计学意义（P〉0．05）,且缓解期仍高于对照组（P〈0．05）;初诊未治AL患儿骨髓中XIAP蛋白水平在急性淋巴细胞白血病（ALL）和急性非淋巴细胞白血病（ANLL）组比较差异无统计学意义（P〉0．05）。结论XIAP蛋白在儿童AL患者中高表达,提示XIAP可能通过抑制肿瘤细胞的凋亡参与了儿童AL的发生、发展。     

血管内皮生长因子在急性白血病中的表达水平及其意义

目的 探讨急性白血病（AL）血清和骨髓单个核细胞培养上清液（CSBMMNC）中血管内皮生长因子（VEGF）表达水平及其临床意义。方法 采用酶联免疫吸附法定量检测71例急性白血病和20例正常对照血清及CSBMMNC中VEGF表达水平。对其中33例初发AL患者采用2个疗程化疗。分析比较VEGF表达水平的变化与疗效的关系。结果ANLL组与ALL组初发患者血清与CSBMMNC中VEGF水平相比，差异无显著性（P〉0．05）。AL初发组、复发难治组分别与完全缓解（CR）组、正常对照组血清及CSBMMNC中VEGF表达水平相比有非常显著性差异（P均〈0．01）。但复发难治组与初发组相比差异无显著性（P均〉0．05）。初发AL血清与CSBMMNC中VEGF水平与骨髓原始细胞数呈显著正相关（r值分别为0．521和0．849，P均〈0．01）。33例初发AL患者采用2个疗程化疗，血清及CSBMMNC中高表达组CR率（36．36％）较低表达组CR率（90．90％）低，有非常显著性差异（P均〈0．01）。结论 动态检测血清及CSBMMNC中VEGF水平可在一定程度上了解AL的状态和体内白血病细胞的总负荷量，对AL的疗效及预后判断有一定的指导意义。     

X染色体相关凋亡抑制蛋白在86例急性白血病中的表达

[目的]探讨急性白血病（AL）患者中X染色体相关凋亡抑制蛋白（XIAP）表达的临床意义。[方法]86例AL患者,其巾初诊未治患者56例,包括44例AML和12例ALL,治疗后完全缓解期（CR）患者22例,缓解后复发患者8例,应用RT～PCR方法检测骨髓细胞中XIAP的mRNA表达。对照组为20名门诊正常人。[结果]比较20例对照组、56例AL初治组、22例AL缓解组和8例AL复发组的XIAPmRNA平均表达水平,差异具有显著性（F=58．77,P〈0．01）;对照组的XIAPmRNA平均表达水平为0．29±0．13,明显低于AL初治患者（0．98±0．30）（P〈0．01）和缓解组（0．45±0．17）（P〈0．05）;缓解组的平均表达水平明显低于初治组（P〈0．01）;复发组的XIAPmRNA平均表达水平最高（1.23±0.30）．不仅明显高于对照组和缓解组（P〈0．01）,也高于初治组（P〈0．05）;XIAP mRNA表达在初治AML和ALL组比较无显著性差异（P=0.21）。[结论]XIAP在急性白血病发病和病情进展中起重要作用,XIAP高表达可能是AL患者预后不良的高危因素之一．     

急性白血病端粒酶活性与p53蛋白表达规律的研究

目的：探讨争性白血病（AL）患者中端粒酶活性与突变型P53蛋白的表达规律,方法：采用聚合酶链反应一酶联免疫吸附法（PCR－ELISA）对端粒酶活性进行定半量测定,用流式细胞仪检测突变型P53蛋白表达,结果：AL患者端粒酶性显著高于对照组＊（P＜0．001）,端粒酶活性在AL的不同病期有统计学差异,其中初治组,复发组均显著高于完全缓解CR组（P＜0．01）,在治疗敏感组与不敏感组间无统计学差异（P＞0．05）,AL患者中突变型P53蛋白阳性率为19．4％（7／36）,在AL的不同病期P53阳性率无统计学差别。在初治敏感组与不敏感组之间亦无统计学差别（P均＞0．05）,端粒酶活性与突变型P53蛋白的表达无关。结论：端粒酶活性与AL的病期和发展密切相关,突变型P53蛋白在AL中表达较低,对临床预后的判断价值较小,端粒酶的激活与P53蛋白表达无相关性。     

急性白血病患者骨髓间隙连接蛋白43的表达及其与耐药的相关性研究

 【摘要】　目的　探讨急性白血病（AL）患者骨髓细胞中间隙连接蛋白43（Cx43）、P-糖蛋白（P-gp）及环氧合酶2（COX-2）基因表达水平及其与AL病程、预后和耐药的关系。方法　77例不同病期AL患者,其中初治36例,完全缓解20例,复发20例,同时以20例异体造血干细胞移植供体及非血液系统恶性病患者为对照。采用SYBR Green实时定量反转录聚合酶链反应（SYBR-RT-PCR）技术,检测骨髓单个核细胞中Cx43、P-gp、COX-2 mRNA的表达,并对37例初治患者进行动态随访。结果　AL初治组Cx43、P-gp、COX-2 mRNA的表达分别为0.52±0.57、1.42±1.06、1.14±0.95,复发组分别为0.20±0.40、2.29±1.11、1.69±0.81,完全缓解组分别为0.95±0.37、0.93±0.73、0.79±0.58,对照组分别为1.16±0.67、0.86±0.63、0.61±0.57。初治、复发AL患者Cx43 mRNA表达水平较对照组及完全缓解组低,差异均有统计学意义（初治组分别P＝0.001、0.005;复发组均P<0.001）;完全缓解组Cx43 mRNA表水平与对照组相比,差异无统计学意义（P＝0.185）。AL患者骨髓细胞液中Cx43 mRNA与P-gp和COX-2 mRNA表达呈负相关,且初治组、完全缓解组及复发组表达均呈负相关（与P-gp r值分别为-0.471、-0.362、-0.526;与COX-2 r值分别为-0.479、-0.344、-0.471）。36例AL初治患者随访4个月,死亡8例,生存28例,死亡患者Cx43 mRNA表达低于生存患者,差异有统计学意义（t＝2.16,P＝0.042）。结论　初治、复发难治AL患者骨髓中Cx43 mRNA的表达下调,同时多药耐药基因P-gp、COX-2 mRNA的表达上调;Cx43过度表达是预后良好因素,Cx43与AL的疗效、预后及化疗耐药密切相关。     

EphA2/EphrinA1在胃癌中的表达及其意义

目的：探计EphA2及其配体EphrinA1在胃癌组织中的表达及与胃癌临床病理特征的关系。方法：利用免疫组织化学法检测82例胃癌手术切除标本的癌组织、癌旁组织及正常胃粘膜内EphA2、EphrinA1和E-cadherin的表达；并采用流式细胞术对其中随机选取的45例标本进行上述三种蛋白的定量分析，采用SPSS软件进行统计学处理。结果：胃癌组织中EphA2、EphrinA1的表达明显高于癌旁组织及正常胃粘膜（P〈0．01），而癌旁组织与正常胃粘膜之间无显著性差异（P〉0．05）；E—cadherin在胃癌组织中的表达明显低于癌旁组织和正常胃粘膜（P〈0．01），而癌旁组织与正常胃粘膜间无显著性差异（P〉0．05）。在胃癌组织中，随分化程度的降低，EphA2、EphrinA1蛋白的表达均显著升高（P〈0．05），而E-cadherin蛋白的表达则显著降低（P〈0．05）；随浸润深度的增加，EphA2、EphrinA1蛋白的表达均显著升高（P〈0．05），而E-cadherin蛋白的表达显著降低（P〈0．05）；淋巴结转移组中EphA2、EphrinA1蛋白的表达显著高于无淋巴结转移组（P〈0．05），而E-cadherin蛋白的表达则显著低于无淋巴结转移组（P〈0．05）；EphA2、EphrinA1和E-cadherin蛋白的表达与肿瘤直径、患者的性别和年龄无关（P〉0．05）EphA2与E—cadherin蛋白的表达呈显著负相关（r=-0．231，P〈0．01），EphrinA1与E-cadherin蛋白的表达呈显著负相关（r=-0．403，P〈0．01），EphA2与EphrinA1蛋白的表达呈显著正相关（r=0．707，P〈0．01）。结论：EphA2／EphrinA1与E-cadherin蛋白表达异常可能共同参与了胃癌的发生、发展与转移；联合捡测三种蛋白对于评价胃癌的恶性程度、判断其转移潜能具有一定的参考价值。     

儿童急性白血病生存素和细胞周期素B1基因的表达

目的：探讨生存素、细胞周期素B1的表达与儿童急性白血病（AL）的关系。方法：采用RT—PCR法对初治或复发AL患儿42例和完全缓解AL（ALCR）患儿13例及非恶性疾病患儿15例骨髓单个核细胞进行基因生存素、细胞周期素B1表达的检测。结果：初治和复发AL组生存素mRNA表达水平分别为0．472和0．810；细胞周期素B1mRNA为0．609和0．739，均高于ALCR组和对照组（均为0）（P〈0．01）；复发AL组生存素mRNA表达水平高于初治组，差异有统计学意义（P=0．025）。生存素基因的表达与白细胞总数及临床分型有关，而与性别、年龄、免疫分型无明显相关性；细胞周期素B1表达与性别、年龄、免疫分型、白细胞总数及临床分型无明显相关关系。儿童AL中生存素与细胞周期素B1的表达呈正相关。生存素mRNA表达水平高者缓解率低。结论：儿童AL中存在生存素、细胞周期素B1异常表达，两者表达存在相关性。生存素是AL复发的高危因素；AL生存素高表达者可以降低AL的化疗敏感性，生存素水平对AL有判断疗效和预后的价值。     

复发/难治急性髓系白血病患者P-糖蛋白的表达及功能分析

 目的 研究复发／难治急性髓性白血病（AML）患者P-糖蛋白表达及其功能。方法 用免疫组化方法检测CD34表达,RT-PCR方法检测P-gp的表达,用流式细胞术检测P-gp的功能。结果 CD34表达与P-gp表达、功能呈正相关（r =0.621,P＜0.001;r =0.496,P＜0.001）,P-gp表达与功能呈正相关（r = 0.574,P＜0.01）;复发／难治组CD34阳性率（66.7 %）高于初治组（32.6 %）（P＜0.05）;复发／难治组P-gp功能阳性率（66.7 %）高于初治组（25.6 %）（P＜0.01）。结论 CD34表达、P-gp表达和P-gp功能三者间呈正相关;复发／难治AL组 CD34阳性率、P-gp功能阳性率高于初治组;P-gp功能的检测更具有临床指导意义。     

食管癌及癌前病变组织中p16蛋白表达的定性和定量分析

目的：定性和定量分析P16蛋白在食管癌及癌前病变组织中的表达，探讨其与食管癌发生发展的关系。方法：应用免疫组化及流式细胞术（FCM），检测P16蛋白在食管癌及癌前病变组织中的表达。结果：1）从鳞状上皮→不典型增生（癌前病变）→原位癌→浸润癌，随着病变加重．P16蛋白表达呈逐渐降低趋势。2）免疫组化结果显示：正常组与不典型增生Ⅰ级组p16蛋白表达差异无显著性（P〉0．05），与其余各组差异有显著性（P〈0．05）；浸润癌组与正常组、不典型增生Ⅰ级组差异均有显著性（P〈0．05）．与其余组差异无显著性（P〉0．05）。3）FCM结果：不典型增生Ⅱ级组与正常组P16蛋白表达量差异有显著性（P〈0．001）．不典型增生Ⅱ级、Ⅲ级组间差异有显著性（P〈0．05）．不典型增生Ⅲ级与食管癌组间差异无显著性（P〉0．05）结论：P16蛋白表达缺失在食管癌变过程早期阶段就已经发生．随着病变发展P16蛋白阳性表达率逐渐降低．其表达缺失可能在食管癌发生、发展及转归过程中起重要作用。     

Irinotecan and radiation in vitro and in vivo

Loss of chromosome sequences at 13q14 (Rbl) and 17p13 (p53) associated with squamous cell carcinoma of head and neck (SCCHN) was evaluated in 12 recurrent tumors and 51 primary tumors from 63 patients. The incidence of LOH at 17p13 was 19 of 50 (38%) tumors, and at 13q14 was 21 of 57 (37%). LOH affecting Rbl and/or p53 was observed in 30 of 63 (48%) SCCHN. Coincident LOH at Rbl and p53 was detected in 10 of 46 (22%) tumors. There were nine cases in which primary and metastatic tumors were obtained from the same patient. Of these, seven were informative and five of these (71%) manifested LOH at p53 in both primary and metastatic sites. Examination of Rbl in these same tumors showed LOH in six of the nine metastases, and of these six, only three revealed LOH in the primary tumor. LOH at p53 or Rbl alone showed no correlation with clinical outcome. However, tumors that manifested LOH at both loci were associated with poorer patient outcome and poorer histological differentiation.     

TCP and NTCP in preclinical and clinical research in Europe

European research in radiation oncology has a long and successful tradition. The aim of this research is to increase the therapeutic window of radiotherapy by increasing the tumor control probability (TCP) and/or by decreasing the normal tissue complication probability (NTCP). This paper summarizes the basic radiobiological concept underlying treatment optimization by TCP-NTCP data and discusses some of the limitations of currently used models. These are controversial in many aspects and cannot be recommended for clinical routine practice but should rather be considered as a research tool.     

Advances in molecular diagnostics and therapeutics in head and neck cancer

Opinion statement Extensive treatment-related morbidities and stagnant survival rates over the past few decades for patients with squamous cell cancer of the head and neck (SCCHN) emphasize the need for novel diagnostics and therapeutics based on the molecular characteristics of the tumor. The development of an early detection test remains largely preliminary. Much attention has recently been given to saliva-based early detection assays that use accepted tumor markers such as p53 and DNA methylation. Most of these studies have focused on feasibility as opposed to prospective clinical trials. To date, early detection saliva assays have failed to yield a high enough sensitivity and specificity for broad population-based screening. The use of saliva as a noninvasive, inexpensive, and accessible diagnostic substrate remains desirable. Unlike SCCHN diagnostics, molecular-targeted therapies for SCCHN will soon be a reality, with many more compounds in the pipeline. The most promising of these drugs target the epidermal growth factor receptor (EGFR), which is known to be overexpressed in squamous cell carcinomas. Cetuximab, a monoclonal EGFR antibody, has shown efficacy in combination with radiotherapy in advanced SCCHN in a recent phase III trial and is currently being petitioned for US Food and Drug Administration approval. Likewise, erlotinib, an EGFR tyrosine kinase inhibitor, has shown favorable results in phase II trials as monotherapy and in combination with chemotherapy. Gefitinib, another EGFR tyrosine kinase inhibitor, has shown efficacy as monotherapy, in combination with chemotherapy, and with chemoradiotherapy. At least two phase III trials of gefitinib in patients with advanced SCCHN are ongoing. Such low-toxicity, tumor-specific targeting strategies will soon be available for patients with head and neck cancer. The challenge is to establish assays to determine which patients are most likely to benefit from these agents.     

Estrogen and Progesterone in Normal Mammary Gland Development and in Cancer

There is emerging evidence that the mammary epithelium in both mice and humans is arranged as a hierarchy that spans from stem cells to differentiated hormone-sensing, milk-producing and myoepithelial cells. It is well established that estrogen is an important mediator of mammary gland morphogenesis and exposure to this hormone is associated with increased breast cancer risk. Yet surprisingly, the primitive cells of the mammary epithelium do not express the estrogen receptor-α (ERα) or the progesterone receptor. This article will review the mammary epithelial cell hierarchy, possible cells of origin of different types of breast tumors, and the potential mechanisms on how estrogen and progesterone may influence the different subcomponents in normal development and in cancer. Also presented are some hypothetical scenarios on how this underlying biology may be reflected in the behavior of ERα(+) and ERα(-) breast tumors.     

Daunorubicin and daunorubicinol pharmacokinetics in plasma and tissues in the rat

Recent evidence suggests that 13-hydroxy metabolites of anthracyclines may contribute to cardiotoxicity. This study was designed to determine the pharmacokinetics of daunorubicin and the 13-hydroxy metabolite daunorubicinol in plasma and tissues, including the heart. Fisher 344 rats received 5 mg kg^–1 daunorubicin i.v. by bolus injection. Rats were killed at selected intervals for up to 1 week after daunorubicin administration for determination of concentrations of daunorubicin and daunorubicinol in the plasma, heart, liver, kidney, lung, and skeletal muscle. Peak concentrations of daunorubicin were higher than those of daunorubicinol in the plasma (133±7 versus 36±2 ng ml^–1;P<0.05), heart (15.2±1.4 versus 3.4±0.4 g g^–1;P<0.05), and other tissues. However, the apparent elimination half-life of daunorubicinol was longer than that of daunorubicin in most tissues, including the plasma (23.1 versus 14.5 h) and heart (38.5 versus 19.3 h). In addition, areas under the concentration/time curves (AUC) obtained for daunorubicinol exceeded those found for daunorubicin in almost all tissues, with the ratios being 1.9 in plasma and 1.7 in the heart. The ratio of daunorubicinol to daunorubicin concentrations increased dramatically with time from <1 at up to 1 h to 87 at 168 h in cardiac tissue. Thus, following daunorubicin injection, cumulative exposure (AUC) to daunorubicinol was greater than that to daunorubicin in the plasma and heart. If daunorubicinol has equivalent or greater potency than daunorubicin in causing impairment of myocardial function, it may make an important contribution to the pathogenesis of cardiotoxicity.