Clinical Significance of CD200 and CD56 Expression in Patients with Acute Myeloid Leukemia

Background: Acute myeloid leukemia (AML) escape from immunosurveillance by immunosuppression. CD200 and CD56 expression represented an independent prognostic factor in many hematological malignancies but its importance in AML patients remains to be identified. Methods: CD200 and CD56 expression were assessed in the bone marrow blasts for Fifty-two (52) newly diagnosed AML by flowcytometry before start of therapy. Results: CD200+ expression was reported in 28.8% of patients while 17.3% of patients showed CD56+ expression. M4 FAB revealed high frequency of both CD200+ and CD56+ expression. The overall survival of CD200+ patients was 19.2% compared to 35.3% in CD200- (P= 0.049). On the other hand, CD56+ patients had the lowest complete remission rate (22.2% vs. 53.4%). In addition, CD56+ population had significant bad influence on overall survival than those of CD56- population (11.1 % vs. 35.5 %, P= 0.047). Conclusions: CD200 and CD56 positive expression by myeloblasts at diagnosis denote poor prognostic indicator and correlated with poor cytogenetic findings. CD200 could be used as therapeutic target in AML.     

Smac/DIABLO与Livin在乳腺癌组织表达及其临床意义

目的：探讨凋亡促进因子Smac/DIABLO和凋亡抑制因子Livin在人乳腺癌组织中的表达及相关性,分析其与乳腺癌转移复发的关系。方法：应用免疫组织化学SP法检测76例乳腺癌组织和相应的癌旁组织中Livin和Smac蛋白的表达情况。结果：76例乳腺癌组织中Livin蛋白表达的阳性率为67.8%（51/76）,显著高于相应的癌旁组织的59.2%（36/76）,P〈0.05;乳腺癌组织中Smac表达为52.6%（40/76）,显著低于癌旁组织的75.0%（57/76）,P〈0.05。两者的表达与组织学分级、肿瘤大小、腋窝淋巴结转移以及术后复发密切相关,P〈0.05。Livin和Smac表达呈显著负相关,γ=-0.238,P〈0.05。结论：Livin的高表达及Smac/DIABLO的低表达或失活可能在乳腺癌的发生、发展中起重要作用。     

Smac/DIABLO与Livin在乳腺癌组织表达及其临床意义

目的:探讨凋亡促进因子Smac/DIABLO和凋亡抑制因子Livin在人乳腺癌组织中的表达及相关性,分析其与乳腺癌转移复发的关系。方法:应用免疫组织化学SP法检测76例乳腺癌组织和相应的癌旁组织中Livin和Smac蛋白的表达情况。结果:76例乳腺癌组织中Livin蛋白表达的阳性率为67.8%(51/76),显著高于相应的癌旁组织的59.2%(36/76),P<0.05;乳腺癌组织中Smac表达为52.6%(40/76),显著低于癌旁组织的75.0%(57/76),P<0.05。两者的表达与组织学分级、肿瘤大小、腋窝淋巴结转移以及术后复发密切相关,P<0.05。Livin和Smac表达呈显著负相关,γ=-0.238,P<0.05。结论:Livin的高表达及Smac/DIABLO的低表达或失活可能在乳腺癌的发生、发展中起重要作用。     

Smac/DIABLO和Survivin shRNA联合转染对大肠癌Lovo细胞凋亡的影响

背景与目的:Smac/DIABLO是一种新发现的促凋亡蛋白,通过阻滞凋亡抑制蛋白(包括Survivin),激活caspase-9和caspase-3而发挥作用.本实验研究线粒体促凋亡蛋白Smac/DIABLO和沉默凋亡抑制基因Survivin联合应用对大肠癌细胞凋亡的影响及其促凋亡机制.方法:构建Survivin的shRNA-EGFP质粒和Smac-pcDNA3.1质粒,将其用脂质体单、共转染至大肠癌Lovo细胞;Western blot检测survivin基因和Smac基因的蛋白表达;Ho-chest 33258染色观察凋亡核形态学变化,并粗略计数凋亡率;流式细胞仪PI单染测凋亡周期;caspase-3活性检测试剂盒检测caspase-3活性.结果:转染48 h后单、共转染组的Smac蛋白水平增加而Survivin蛋白水平下降;单、共转染组Hochest 33258染色可见明显的凋亡核形态学变化,凋亡率高于各对照组,且共转染组凋亡率(18.5±1.7)%高于单转染组(9.6±1.8)%、(15.0±0.3)%;Smac组G0/G1期增多为(51.0±6.2)%,而Survivin shRNA组S期细胞增多为(53.3±1.3)%(P＜0.05);caspase-3活性比Smac和Survivin shRNA共转组为(169±3)%,高于Smac组(143±5)%、Survivin shRNA组(152±6)%,且都高于各对照组(P＜0.05).结论:Smac/DIABLO和Sur-vivin shRNA协同作用激活caspase-3途径,抑制大肠癌Lovo细胞生长而促进细胞凋亡,Smac阻滞细胞周期于G0/G1期而RNA干扰Survivin后细胞周期阻滞于S期.     

急性髓系白血病患者HOX11基因的表达及意义

目的 探讨HOX11基因在急性髓系白血病(acute myeloid leukemia,AML)中的表达及对预后的影响,为个体化治疗提供依据.方法 应用多重巢式RT-PCR方法对73例初诊AML患者的融合基因进行检测,对HOX11基因表达阳性和阴性的患者进行标准治疗后的疗效进行分析.结果 在预后良好组、预后中等组及预后不良组AML患者的标准治疗中,HOX11基因阳性表达患者的第一疗程完全缓解率(complete remission rate)并不高于阴性表达的患者(P＞0.05),而复发或死亡率(relapse or mortality rate)与HOX11基因阴性表达的患者比较差异有统计学意义(P＜0.05).结论 HOX11基因的表达可能影响AML患者的预后.     

Expression of N-CAM (CD56) on Acute Leukemia Cells: Relationship with Disease Chracteristics and Outcome

Expression of CD56 was analyzed by indirect immunofluorescence method on bone marrow samples from 94 newly diagnosed patients with acute leukemia (AL), including 59 acute myelogenous leukemias (AML) and 35 acute lymphoblastic leukemias (ALL). CD56 was expressed on 17 f 18% (range: 0-728) of AML cells and 24 f 248 (range: 0-988) of ALL cells. without significant diffcrences between FAB subtypes in AML, nor immunologic subtypes in ALL. Expression of CD56 was not associated with any clinical or biological characteristic at diagnosis, nor with prognosis in AML or ALL. We do not confirm previously described relationships between CDS6 expression and initial characteristics and evolution of acute leukemia.     

Osteopontin b and c isoforms: Probably Unique Molecular Candidates Associated with Leukemic Stem Cells Chemoresistance in Acute Myeloid Leukemia

Despite impressive advances in the therapeutic approches, the long-term survival rate of acute myeloid leukemia (AML) is considered to be significantly low as a result of resistance to the treatment and desease relapse. Among multitude oncogenic proteins invovlved in aquisition of chemo-resistance phenotype, osteopontin (OPN) recently attracted tremendous attentions. In spite of the well-defined association between OPN expression and cure rate in solid tumors, there is a scarcity of analysis on the role of this protein in AML. Based on the critical role of OPN in cell survival, it seems reasonable to hypothesize that the high expression of OPN isoforms may participate in disrupting the regulation of apoptosis in AML cells and it s relapse. To investigate the association between induction of apoptosis and OPN isoform expression, two distinct AML cell lines (KG-1 as a leukemic stem cell model and U937) were treated with chemotherapy drugs and the cell viability and apoptosis were evaluated by MTT and annexin/PI assay. After determination suitable drugs doses, the mRNA expression level of OPN and OPN-related genes were investigated. Our results demonstrated for the first time that the acquired up-regulation of OPN-b and c isoforms might prevent conventional chemotherapy regimen-induced apoptosis in AML cells. Moreover, the resulting data revealed that up-pregulation of OPN-b and c in AML cells was concurrently associated with the up-regulation of AKT, VEGF, CXCR4, STAT3 and IL-6 expression. To sum up, this study suggest that OPN-b and c isoforms could be considered as a unique beneficial molecular biomarker which is associated with chemoresistance. Hence, this isoforms are considerable as a potential moleculare candidate for detection of minimal residual disease (MRD) and determination of remission in AML patients. Furthure evaluation with quantative Real tim PCR on patient sample to comfim this finding seems to be be nessesary.     

Prospective Monitoring of Minimal Residual Disease in Acute Myeloid Leukemia with Inversion(16) by CBFβ/MYH11 RT-PCR: Implications for a Monitoring Schedule and for Treatment Decisions

Minimal residual disease in patients with acute myeloid leukemia (AML) with inversion(16) can be monitored by CBFβ/MYH11 RT-PCR. While the association between molecular remission (MR) in bone marrow (BM) and peripheral blood (PB) and long-term clinical remission (CR) seems to be established, there are insufficient data on the kinetics of CBFβ/MYH11. We have performed a prospective study in order to generate a reasonable and sufficient schedule for PCR-monitoring. 11 patients with AML and inversion (16) in complete hematological remission have been prospectively monitored by CBFβ/MYH11 RT-PCR in their BM and PB during an observation period of 7 to 67 months (median 32 months). Patients were followed during consolidation chemotherapy with repetitive cycles of high-dose Ara-C and after autologous or allogeneic stem cell transplantation in 2^nd CR or refractory AML. MR never coincided with achievement of CR but occurred between 2 and 8 months after hematological remission. All patients in continuous CR were PCR-negative after 1-8 (median 4) months. Two patients relapsed despite MR for 10 to 15 months. Molecular relapse preceded hematological relapse by 3 to 5 months. Three out of four patients who were not in MR after 8 months relapsed. Allogeneic stem cell transplantation was able to eradicate minimal residual disease in 4/4 patients. In 2 patients a temporary reconversion to PCR-positivity was reversed by reduction of immunosuppression. 1 patient did not become PCR-negative until compete withdrawal of immunosuppression. We suggest that BM and PB should be examined after the last consolidation treatment. In case of MR, PB should be examined every 1 to 2 months and BM examination should be done only in case of PCR-positivity in PB in order to confirm the molecular relapse and to identify an impending cytogenetic and/or hematological relapse. CBFβ/MYH11 RT-PCR monitoring is able to predict relapse 3 to 5 months prior to overt hematological relapse, offers a window of opportunity for preemptive therapy of molecular relapse and confers implications for immunotherapy in the setting of allografting.     

全反式维甲酸与亚砷酸联合柔红霉素对NB4细胞CD11b表达的影响

 目的　观察全反式维甲酸（ATRA）与亚砷酸（ATO）单独以及二药联合柔红霉素（DNR）诱导分化治疗过程中对NB4细胞CD11b表达的影响,及其对高白细胞血症、急性早幼粒细胞白血病（APL）分化综合征的作用。方法　采用荧光素标记的CD11b单克隆抗体,流式细胞术动态检测各组药物作用于NB4细胞24、48、72、168 h后细胞CD11b的表达状况。结果　1 μmol／L ATRA作用于NB4细胞后,CD11b表达随作用时间的延长逐渐增加,呈时间依赖性。24、48、72、168 h CD11b表达分别为（33.34±3.15）%、（55.59±5.13）%、（86.08±5.12）%、（90.69±2.69）%,在同一时间点均高于空白对照组（P＜0.01）。1 μmol／L ATO作用于NB4细胞后,随时间延长CD11b表达亦逐渐增加,但各时间点之间CD11b表达差异无统计学意义;在同一时间点CD11b表达低于ATRA组（P＜0.01）,但与空白对照组比较差异无统计学意义（P＞0.05）。1 μmol／L ATRA+1 μmol／L ATO作用于NB4细胞后,24、48 h CD11b表达分别为（16.92±1.05）%、（17.01±0.22）%,与空白对照组比较差异无统计学意义（P＞0.05）,72、168 h CD11b表达高于空白对照组（P＜0.05）,分别为（18.81±1.40）%、（25.61±4.54）%;但在同一时间点CD11b表达均低于ATRA组（P＜0.01）。1 μmol／L ATRA+1 μmol／L ATO+1 μmol／L DNR作用于NB4细胞后,在同一时间点CD11b表达低于ATRA组（P＜0.01）;与空白对照组比较差异无统计学意义（P＞0.05）。结论　ATRA联合ATO或加用DNR诱导治疗APL,可能由于避免了APL细胞诱导分化过程中CD11b表达的增高,从而在一定程度上减少了高白细胞血症、APL分化综合征的发生。     

CD56+表达对急性单核细胞白血病儿童的临床特征、疗效及预后的影响

目的探讨CD56⁺表达对急性单核细胞白血病(AML-M5)儿童临床特征、疗效及预后的影响。方法对84例14岁以下AML-M5患儿临床资料进行回顾性分析,分析CD56的表达与临床特征、疗效及预后的关系。结果CD56⁺表达率为32.14%,白细胞计数(73.21±12.93)×10⁹/l,高白细胞血症占51.85%,髓外浸润占59.26%;CD56^-表达率为67.86%,白细胞计数(54.21±10.64)×10⁹/l,高白细胞血症占15.79%,髓外浸润占29.82%,比较差异具有统计学意义(P<0.05)。CD56+患儿治疗完全缓解率为62.96%、死亡率为7.41%,CD56^-患儿完全缓解率66.67%、死亡率8.77%,比较差异无统计学意义(P>0.05);CD56⁺达完全缓解的患儿随访期间复发率76.47%、持续缓解率为23.53%,平均生存时间(10.92±4.71)个月,CD56^-患儿复发率28.95%、持续缓解率为71.05%、平均生存时间(15.15±8.12)个月,比较差异具有统计学意义(P<0.05)。结论CD56⁺表达的14岁以下AML-M5患儿高白细胞血症、髓外浸润发生及复发率较高,平均生存时间较短。CD56表达能够为患儿的治疗及预后提供指导信息。     

Head and neck cancer relapse after chemoradiotherapy correlates with CD163+ macrophages in primary tumour and CD11b+ myeloid cells in recurrences

Background:

We investigated the prognostic role of tumour-associated macrophages (TAMs) in patients with head and neck squamous cell carcinoma (HNSCC) treated with definitive chemoradiotherapy (CRT).

Methods:

The expression of CD68+, CD163+ and CD11b+ cells was assessed using immunohistochemistry in n=106 pre-treatment tumour biopsy samples and was correlated with clinicopathological characteristics, including T-stage, N-stage, grading, tumour localisation, age and sex as well as local failure-free survival (LFFS), distant metastases-free survival (DMFS), progression-free (PFS), and overall survival (OS). Finally, TAMs expression and vessel density (CD31) were examined in n=12 available early local recurrence samples and compared with their matched primary tumours . The diagnostic images and radiotherapy plans of these 12 patients were also analysed. All local recurrences occurred in the high radiation dose region (⩾70 Gy).

Results:

With a median follow-up of 40 months, OS at 2 years was 60.5%. High CD163 expression in primary tumours was associated with decreased OS (P=0.010), PFS (P=0.033), LFFS (P=0.036) and DMFS (P=0.038) in multivariate analysis. CD163 demonstrated a strong prognostic value only in human papillomavirus (p16^INK4)-negative patients. Early local recurrence specimens demonstrated a significantly increased infiltration of CD11b+ myeloid cells (P=0.0097) but decreased CD31-positive vessel density (P=0.0004) compared with their matched primary samples.

Conclusions:

Altogether, baseline CD163 expression predicts for an unfavourable clinical outcome in HNSCC after definitive CRT. Early local recurrences showed increased infiltration by CD11b+ cells. These data provide important insight on the role of TAMs in mediating response to CRT in patients with HNSCC.     

以CD56+小肠粒细胞肉瘤为首发表现的急性单核细胞白血病1例并文献复习

目的 探讨粒细胞肉瘤的发病机制、诊治与转归。方法分析1例以CD56+小肠粒细胞肉瘤为首发表现的急性单核细胞白血病患者的临床病理特征及实验室检查结果,并复习相关文献。结果 收治1例男性患者,39岁。小肠系膜切除物免疫组化检测示：粒细胞肉瘤,CD56阳性;骨髓细胞学检查示：急性单核细胞白血病,诊断为急性单核细胞白血病伴小肠粒细胞肉瘤。经IA方案诱导化疗后疾病达完全缓解,继续IA方案巩固化疗,拟行异基因造血干细胞移植。结论以小肠粒细胞肉瘤为首发表现的急性白血病较为少见。粒细胞肉瘤的预后差,CD56抗原阳性提示预后不良,经诱导化疗获疾病缓解的患者应尽快行异基因造血干细胞移植以期达到长期生存。     

Prognostic relevance of CD56 expression in multiple myeloma: a study including 107 cases treated with high-dose melphalan-based chemotherapy and autologous stem cell transplant

CD56 is a neural adhesion molecule and expressed in 70-80% cases of multiple myeloma (MM). Lack of CD56 expression has shown to be a poor prognosis in MM patients treated with conventional chemotherapy, but its prognostic relevance in MM treated with high dose chemotherapy and autologous stem cell transplant (ASCT) is not known. CD56 expression was evaluated by immunohistochemistry on bone marrow paraffin embed specimens from 107 MM cases undergoing Melphalan-based high dose therapy and ASCT. CD56 was expressed by the myeloma cells in 71% of the patients. CD56 negative myeloma was associated with bone lesions ( p = 0.032), but there was no association with any other biological or genetic risk factors including deletions 13q, p53 and IgH translocations, as evaluated by fluorescence in situ hybridization (FISH). There was no significant difference between CD56 positive and CD56 negative myeloma for progression free or overall survival ( p = 0.28 and p = 0.67, respectively). In contrast to reports of CD56 in myeloma treated with conventional chemotherapy, CD56 negativity was not found to confer a poor prognosis in these patients, suggesting Melphalan-based high-dose chemotherapy and ASCT may overcome the adverse influence of CD56 negative myeloma.     

CD82/KAI1基因蛋白在62例胃癌中的表达

[目的]探讨胃癌组织中抑癌基因CD82/KAI1蛋白的表达与胃癌临床生物学行为及预后的关系。[方法]采用免疫组织化学方法检测62例胃癌原发灶、阴性组织切缘中CD82/KAI1基因蛋白产物的表达。[结果]胃癌组织中KAI1蛋白阳性表达率为72.5%,显著低于阴性组织切缘的95.2%（P〈0.01）。KAI1蛋白在浸润深度的T3＋T4组（65.1%）较T1＋T2组（89.5%）;低分化组（20.5%）较高、中分化组（88.9%）;淋巴结转移组（59.1%）较无淋巴结转移组（94.4%）;TNM分期中的Ⅲ～Ⅳ期组（63.4%）较Ⅰ～Ⅱ期组表达（90.5%）均降低（P〈0.05）。胃癌组织中的KAI1蛋白表达与患者性别、年龄、肿瘤大小、Lauren分型无关,胃癌患者生存期5年以下组的胃癌组织中KAI1蛋白的表达水平明显低于生存期5年以上组（P〈0.05）。[结论]胃癌组织中KAI1基因蛋白表达降低与胃癌的侵袭、转移及临床病理进展有关。     

Importance of FISH combined with Morphology,Immunophenotype and Cytogenetic Analysis of Childhood/ Adult Acute Lymphoblastic Leukemia in Omani Patients

Genetic changes associated with acute lymphoblastic leukemia (ALL) provide very important diagnostic andprognostic information with a direct impact on patient management. Detection of chromosome abnormalities byconventional cytogenetics combined with fluorescence in situ hybridization (FISH) play a very significant rolein assessing risk stratification. Identification of specific chromosome abnormalities has led to the recognition ofgenetic subgroups based on reciprocal translocations, deletions and modal number in B or T-cell ALL. In the lasttwelve years 102 newly diagnosed childhood/adult ALL bone marrow samples were analysed for chromosomalabnormalities with conventional G-banding, and FISH (selected cases) using specific probes in our hospital.G-banded karyotype analysis found clonal numerical and/or structural chromosomal aberrations in 74.2% ofcases. Patients with pseudodiploidy represented the most frequent group (38.7%) followed by high hyperdiploidygroup (12.9%), low hyperdiploidy group (9.7%), hypodiploidy (<46) group (9.7%) and high hypertriploidygroup (3.2%). The highest observed numerical chromosomal alteration was high hyperdiploidy (12.9%) withabnormal karyotypes while abnormal 12p (7.5%) was the highest observed structural abnormality followed byt(12;21)(p13.3;q22) resulting in ETV6/RUNX1 fusion (5.4%) and t(9;22)(q34.1;q11.2) resulting in BCR/ABL1fusion (4.3%). Interestingly, we identified 16 cases with rare and complex structural aberrations. Application ofthe FISH technique produced major improvements in the sensitivity and accuracy of cytogenetic analysis withALL patients. In conclusion it confirmed heterogeneity of ALL by identifying various recurrent chromosomalaberrations along with non-specific rearrangements and their association with specific immunophenotypes. Thisstudy pool is representative of paediatric/adult ALL patients in Oman.     

CD34 is not Expressed by Blasts in a Third of B-ALL Patients and its Negativity is associated with Aberrant Marker Expression: A Retrospective Analysis

Background: CD34 antigen is expressed by early hematopoietic progenitor cells and acute leukemia cells. Its expression is associated with good prognosis in acute myeloid leukemia. Literature is sparse on its prognostic significance in B- acute lymphoblastic leukemia (B-ALL) especially from India. Hence the present study was undertaken to analyse the frequency of CD34 expression in B-ALL in Indian patients and determine its prognostic significance by associating with other prognostic markers and aberrant antigen expression. Methods: Seventy-five B-ALL patients diagnosed by flow cytometry over a period of 3½ year were studied. Correlation of CD34 expression was studied with gender, age, total leucocyte count (TLC), French-American-British (FAB) morphological type, immuno-phenotypic markers, cytogenetics and minimal residual disease. Differences between groups were evaluated using Student’s T-test for quantitative data and Chi-square test/Fishers exact T-test for qualitative variables. P value