急性髓性白血病bcl—2mRNA表达与抗癌药敏感性及临床疗效关系

目的：探讨急性髓性白血病bcl-2 mRNA表达与抗癌药物敏感性及临床疗效的关系。方法：用半定量RT－PCR方法检测29例急性髓性白血病bcl-2 mRNA表达，并用MTT法测定10种抗癌药的抑制率，以正常健康人bcl-2mRNA表达水平均值作为分界点，分成阴性组和阳性组，统计比较抗癌药在两组中的抑制率并观察临床疗效。结果：bcl-2mRNA表达阴性组抗癌药抑制率大于阳性组，其中氮芥，阿糖胞苷，鬼臼乙叉甙存在显差异（P＜0．05），甲氨蝶呤，环磷酰胺存在极显差异（P＜0．01）。bcl-2mRNA阴性组化疗缓解率高于阳性组，两组具统计学意义（P＜0．05）。结论:bcl-2 mRNA在急性髓性白血病表达可作为一种多药抵抗分子来预测体外药物的敏感性,bcl-2mRNA低表达化疗疗效好。     

bcl—2基因在乳腺癌中的表达及临床意义

目的研究bcl-2基因在乳腺癌癌灶、癌旁组织、转移淋巴结及正常组织中的表达,与肿瘤形成、发展的关系及临床意义.方法应用相对定量RT-PCR方法,检测bcl-2基因在16例乳腺癌癌灶、癌旁组织、转移淋巴结及正常乳腺组织中的表达.结果乳腺癌组织中bcl-2 mRNA表达相对值(0.264±0.137),明显高于正常乳腺组织表达相对值(0.033±0.031)(P<0.005);转移淋巴结bcl-2 mRNA表达相对值(0.272±0.076),癌旁组织bcl-2mRNA表达相对值(0.131±0.083),明显高于正常乳腺组织表达相对值(P<0.05);淋巴结转移之癌灶组bcl-2 mRNA表达相对值(0.381±0.096)明显高于非转移之癌灶组(0.186±0.102)(P<0.05).结论bcl-2基因的高表达可能在乳腺癌的发生、发展中起一定的作用.     

High expression of bcl-2 mRNA as a determinant of poor prognosis in acute myeloid leukemia

Background: The bcl-2 oncoprotein is suggested to be directly involved in the emergence of drug resistance by disrupting or delaying the apoptotic program and promoting tumor survival.Patients and methods: In order to define the clinical relevance of the bcl-2 mRNA expression in acute myeloid leukemia (AML) and its correlation to therapy outcome and prognosis, we analyzed 219 AML bone marrow (BM) samples, including 119 patients with de novo AML at presentation, 37 with AML following myelodysplastic syndrome (MDS), as well as 42 BM samples of AML in relapse and 21 in complete remission (CR) using RT-PCR. For performing quantitative measurements of bcl-2 mRNA, we developed a quantitative RT-PCR.Results: Bcl-2 mRNA was detectable in 133 of 156 (84%) patients at diagnosis and 40 of 42 (95%) at relapse. AML patients with high bcl-2 mRNA expression achieved lower CR rates than those with no or low expression. Concerning the long-term outcome, the overall (OS) and disease-free survival (DFS) was significantly worse in AML patients with high expression levels of bcl-2 mRNA. The three-year OS for all newly diagnosed AML patients was 49% and 10% (P = 0.028), respectively, and 71% and 15% (P = 0.0004) for patients <60 years. Comparable significant differences were observed for the DFS.In AML following MDS and patients >60 years, the bcl-2 expression was not associated with remission rate or survival.Conclusions: The expression of bcl-2 mRNA may serve as a prognostic factor predicting remission outcome and long-term prognosis in AML.     

Prognostic relevance of circulating matrix metalloproteinase-2 in acute myeloid leukaemia patients

Matrix metalloproteinases (MMPs) were postulated to have important implication in progression and invasiveness of many malignant disorders. On the other hand the biological role of MMP-2 in acute myeloid leukaemia (AML) is not fully clear. Serum samples from 37 adult patients with AML had been taken before chemotherapy was administered. In addition 20 out of the 37 patients were analysed again after achieving complete remission (CR). Ten samples from healthy volunteers were evaluated as the control. Total MMP-2 levels were measured using ELISA Kit obtained from R&D system. MMP-2 serum levels were significantly lower in pretreatment AML patients than that in the normal controls (p = 0.000) and in CR (p = 0.007). No significant correlations were detected between pretreatment sMMP-2 levels and FAB subtypes, peripheral blood blast cell counts, peripheral blood WBCs, bone marrow blast cell counts or blast cell distribution ratio. The prognostic value of MMP-2 was evaluated by dividing AML patients into high and low MMP-2 groups using the pretreatment median MMP-2 level of the AML group as the cut-off. The authors found that patients in the high group survived for a significantly shorter time than those patients in the lower MMP-2 group. High pretreatment levels of sMMP-2 among AML patients were associated with poor survival. Prospective studies are recommended to establish the clinical value of longitudinal sMMP-2 measurement.     

急性髓性白血病患者LRP、MRP基因表达

目的：研究急性髓性白血病ＬＲＰ基因、ＭＲＰ基因表达与疗效的关系，方法：用ＲＴ－ＰＣＲ法检测３７例急性髓性白血病ＬＲＰ基因、ＭＲＰ基因表达，按此两种基因表达分成ＬＲＰ－／ＭＲＰ－、ＬＲＰ－ＭＲＰ＋、ＬＲＰ＋／ＭＲＰ－、ＬＲＰ＋／ＭＲＰ＋共４组，统计比较各组疗效。结果：与ＬＲＰ－／ＭＲＰ－组比较，其余３组疗效较低，其中ＬＲＰ＋／ＭＲＰ－组、ＬＲＰ＋／ＭＲＰ＋组存在显著差异，但ＬＲＰ＋／ＭＲＰ＋组与ＬＲ     

安吖啶诱导白血病细胞程序化死亡及bcl—2表达的研究

目的探讨安吖啶诱导不同分化阶段的人急性粒白血病细胞株ＨＬ－６０（ＡＭＬ２）及ＮＢ４细胞（ＡＭＬ３）程序化死亡的作用。方法应用光镜、ＤＮＡ电泳及流式细胞术检测程序化死亡细胞,应用免疫组化法检测ｂｃｌ－２基因表达。结果１～２０μｇ／ｍｌ的安吖啶作用６～１２小时后,ＮＢ４细胞的形态学改变较ＨＬ－６０细胞更明显;ＮＢ４细胞的ＤＮＡ梯带较ＨＬ－６０细胞明显;流式细胞术检测ＮＢ４程序化死亡细胞数较ＨＬ－６０细胞多;ＨＬ－６０细胞ｂｃｌ－２蛋白的平均水平明显高于ＮＢ４细胞。结论ＨＬ－６０细胞对安吖啶诱导程序化死亡作用的敏感性较低,可能与ｂｃｌ－２基因高表达有关。     

Auer rods in the acute myeloid leukemias: frequency and methods of demonstration

One hundred and twenty-five cases of acute myeloid leukemia (AML) were reviewed for the presence of Auer rods by two peroxidase methods. The percentage of Auer rods recognized by Wright-Giemsa (WG) staining was 20.8% but three times higher by peroxidase staining techniques using either benzidine base as a substrate (PO method) or 3,3'-diaminobenzidine as a substrate (DAB method). Both PO and DAB methods were equally sensitive in identifying Auer rods, the higher percentage being in FAB types M2 and M4 and lower in M1, M3, and M5B, with none in M5A. Cell counts of at least 100 cells were as accurate as counting cells up to a total of 1000 consecutive blasts.     

Modulation by bexarotene of mRNA expression of genes in mouse lung tumors

Bexarotene has demonstrated chemopreventive and therapeutic efficacy towards mouse lung tumors. Using specimens from our published study that demonstrated the efficacy of bexarotene, we report herein its ability to modulate mRNA expression of genes in both lung and lung tumors. Strain A/J mice were administered vinyl carbamate to induce lung tumors. This was followed by 200 mg/kg body weight of bexarotene administered by oral gavage during Wks 4-25 or 23-25. The mice were sacrificed at Wk 25. The expression of 26 genes was decreased in lung tumors, whereas only two genes, Apolipoprotein D and CYP26b, had their mRNA expression increased by bexarotene. Genes with increased mRNA expression in untreated lung tumors include: epiregulin and kininogen-1 (increased by more than 40-fold) and Caspase-3, Cyclin D1, DNA methyltransferase 3a (Dnmt-3a), E-prostanoid 3 receptor (EP3), c-myc, surfactant protein-C, and survivin (increased by 1.7- to 3.6-fold). Bexarotene decreased the mRNA expression of Caspase-3, Dnmt-3a, EP3, and survivin, as well as the expression of the Cyclin E1, estrogen receptor-alpha, and iNOS genes. Bexarotene had a greater effect in decreasing the expression of Caspase-3, Cyclin E1, Dnmt-3a, EP3, iNOS, and survivin, when administered to mice with established tumors than when administered to mice while tumors were emerging. In summary, bexarotene modulated mRNA expression of genes in mouse lung tumors, being more effective in established tumors than in emerging tumors, suggesting that modulation of expression could be useful as a biomarker for the therapeutic and chemopreventive activity of the drug, especially in established tumors.     

Monitoring of minimal residual disease in acute myeloid leukemia

Two highly sensitive methods, multiparameter flow cytometry (MFC) and real-time quantitative PCR (RQ-PCR), are increasingly used to monitor minimal residual disease (MRD) and to guide risk-adapted management in acute myeloid leukemia (AML). An independent prognostic impact has been demonstrated for MRD levels obtained by both methods. MFC has been found particularly useful for assessment of early clearance of malignant cells and after consolidation therapy. At the latter checkpoint, MRD levels quantified by RQ-PCR in AML with fusion genes also have the strongest prognostic power. In addition, highly predictive initial expression levels have been identified by RQ-PCR. Both methods are capable of early detection of relapse. Through the use of all available markers including NPM1 mutations and FLT3 mutations in addition to fusion genes, RQ-PCR-based MRD assessment is possible in more than half of patients, whereas MFC is applicable to most AML cases. With a sensitivity of 10(-4) (PML-RARA) to 10(-7) (patient-specific primers, FLT3 and NPM1 mutations), RQ-PCR is more sensitive in most cases. Large clinical trials will determine the exact role and place of immunologic and RQ-PCR-based monitoring of MRD in the therapy of patients with AML.     

CD34抗原、bcl-2蛋白表达与急性髓性白血病化疗耐药

目的　检测急性髓性白血病 (AML)中CD34 ,bcl 2的表达 ,并评价其临床意义及CD34 ,bcl 2间可能存在的关系。方法　免疫细胞化学APAAP法。结果　CD34 、bcl 2表达与AML外周血WBC数。骨髓原始细胞比例、CUF L等临床特征无关 ;而继发性AMLCD34 水平显著高于原发性AML患者 (P <0 .0 5 ) ;但bcl 2水平在原发 ,继发AML中无差别。原发性AML中CD34 水平与化疗疗效无关 (P >0 .0 5 ) ;无论原发AML或AML中 ,bcl 2表达均与化疗耐药有关。结论　CD34 并非一个能独立耐药相关的因素 ,而bcl 2可以独立决定化疗反应 ;CD34 和bcl 2无相关性。     

鼻咽癌细胞bcl—2基因异常与EB病毒感染的关系

目的 研究鼻咽癌细胞bcl－2基因异常与EB病毒感染的关系。方法 采用半套式原位聚合酶链反应和免疫组织化学技术对41例鼻咽癌细胞中bcl－2基因重排、bcl－2蛋白表达率为82．92％（34／41）、LMP－1的阳必率为51．2％（21／41）,bcl－2基因异常（重排和蛋白异常表达）与LMP－1的阳性率无显著性差异（P＜0．05）,bcl－2／JH融合基因形成与bcl－2蛋白表达无显著性差异（P＜0．05）。结论 bcl－2基因在鼻咽癌发生过程中可能起一定作用。mbr和mcr区域同为bcl－2基因在鼻咽癌的2个重要断裂区;bcl－2基因重排可能不是引起bcl－2蛋白异常表达惟一原因;EBV感染对bcl－2基因重排和bcl－2蛋白过表达可能无影响。     

Prognostic value of ERBB family mRNA expression in breast carcinomas

The ErbB-driven autocrine growth pathway has been implicated in the development and progression of most common human epithelial malignancies; its blockade is therefore a promising therapeutic strategy, and several candidate drugs are currently undergoing clinical trials. Paradoxically, little is known of the expression pattern of these 4 genes in human tumors, and the clinical significance of the 2 most recently discovered ERBB genes, ERBB3 and ERBB4, is unclear. We used a real-time quantitative RT-PCR assay to quantify ERBB family mRNA copy numbers in a large series of breast tumors from patients with known long-term outcome. ERBB gene expression varied widely, by more than 2 orders of magnitude for ERBB1 and ERBB3, more than 3 orders for ERBB2 and more than 4 orders for ERBB4. We found a positive correlation between ERBB3 and ERBB4 mRNA levels, and a negative correlation between the expression of these 2 latter genes and that of ERBB1. Compared to normal breast tissue, ERBB1 was underexpressed (82.3% of tumors), ERBB2 (16.9%) and ERBB3 (46.2%) were overexpressed and ERBB4 was both underexpressed (24.6%) and overexpressed (29.2%). Links were also found between ERBB status on the one hand and Scarff-Bloom-Richardson (SBR) histopathological grade and estrogen receptor alpha (ERa) status on the other hand. Relapse-free survival (RFS) was shorter among patients with ERBB3-overexpressing tumors (p=0.0092) and longer among those with ERBB4-underexpressing tumors (p=0.0085) relative to patients with normal expression of the respective genes; in contrast, RFS was not significantly influenced by ERBB1 or ERBB2 mRNA status. Only ERBB4 status retained prognostic significance in Cox multivariate regression analysis (p=0.015). Our results point to the involvement of several ErbB-specific ligands (amphiregulin and neuregulin 1) and enzymes or adaptor molecules (PI3K, Src, Shc and Grb7) in the ErbB pathway dysregulation associated with breast cancer. These findings reveal a complex expression pattern of ERBB gene family members in breast tumors and suggest that it is this pattern of expression, rather than the expression of individual family members, that should be taken into account when evaluating antitumoral drugs designed to target these receptors.     

Expression in bladder transitional cell carcinoma by real-time quantitative reverse transcription polymerase chain reaction array of 65 genes at the tumor suppressor locus 9q34.1-2: identification of 5 candidates tumor suppressor genes

Frequent deletions on 9q34.1-2 were reported in bladder transitional cell carcinoma. High deletion mapping studies delimited a critical interval between markers D9S61 and D9S66, which is highly susceptible to contain a tumor suppressor gene. Expression level of the 65 genes localized in this region was analyzed by real-time quantitative RT-PCR, comparing tumor to normal urothelium. Five genes exhibited a significantly reduced expression level: C9orf9, KIAA0625, ABL1, LAMC3 and KIAA1857-netrin-G2, which exhibited the most significant downregulation (p=0.0007). KIAA1857-netrin-G2 belongs to the netrins and might then be a tumor suppressor gene in bladder cancer, as netrin1 receptor DCC has been implicated in tumorigenesis.     

Relevance of renin expression by real-time PCR in acute myeloid leukemia

The search for useful molecular markers in the diagnosis of AML and in the follow-up of minimal residual disease (MRD) has been the focus of many recent studies. Previous research showed that, while normal bone marrow cells lack expression of renin, myeloid blasts have been reported to do so. The aim was to study the expression of the renin gene by the use of real-time quantitative PCR (RQ-PCR) at diagnosis in acute myeloid leukemia patients (AML) and to assess its possible relevance in the prognosis and outcome of such patients. This study analysed 76 samples from patients with AML, with follow-up of positive patients. Thirty-one patients (41%) were positive for renin gene expression at diagnosis. All renin-positive patients at diagnosis showed no expression during complete remission (CR), but expression recurred in those experiencing relapse and persisted when the disease was refractory to treatment. Although the results suggest that the sub-group of renin-positive AML patients might have a worse outcome and a higher relapse rate (at 5 years, the projected rate of disease-free survival was 18.5 ± 9.8% for renin-positive and 23.5 ± 8.8% for renin-negative patients), no significant differences were found. It is believed that further studies should aim to validate whether such a difference exists, using a much larger and homogeneus group of patients.