前列腺癌组织中TMPRSS2基因与ETS家族基因的多种融合亚型及意义

目的：了解前列腺癌组织标本中跨膜丝氨酸蛋白酶2（TMPRSS2）基因与ETS转录因子家族成员ETS相关基因（ERG）、ETS变异体1（ETV1）及ETS变异体4（ETV4）基因之间的融合情况及意义。方法：采用巢式逆转录聚合酶链反应（RT—PCR）琼脂糖凝胶电泳法，检测32例前列腺癌患者及34例前列腺良性增生患者，前列腺组织中TMPRSS2基因与ETS家族基因融合的TMPRSS2／ERG、TMPRSS2／ETV1、TMPRSS2／ETV4转录体；琼脂糖凝胶电泳阳性者，纯化PCR产物进行直接测序，用BLAST在线软件比对确定融合位点；分析融合基因与Gleason分级关系。结果：在32例前列腺癌患者组织标本中，检测到TMPRSS2／ERG融合基因17例（53．1％），含5种不同融合基因亚型，其中1种为新发现融合基因亚型（Genbank登录号：EU090248），单一标本中可检测到一种以上TMPRSS2／ERG融合基因亚型；检测到TMPRSS2／ETV1融合基因2例（6．3％），为新发现融合基因亚型（Genbank登录号：EU090249）；未检到TMPRSS2／ETV4融合基因型；34例前列腺良性增生组织标本中均未检测到TMPRSS2／ERG、TMPRSS2／ETV1和TMPRSS2／ETV4融合基因型；按Gleason评分值分为中分化与低分化的两组前列腺癌组织标本之间融合基因阳性率无统计学差异（P=0．169）。结论：前列腺癌组织中存在TMPRSS2／ERG和TMPRSS2／ETV1融合基因及多种亚型；前列腺癌融合基因的发现有望为前列腺癌的发病机制研究提供新的思路。     

RT-PCR法检测EWS-FLI1融合基因诊断尤文肉瘤/外周原始神经外胚层瘤

目的探讨尤文肉瘤/外周原始神经外胚层瘤(EWS/pPNET)石蜡包埋组织中EWS-FLI1融合基因表达的临床病理意义。方法采用免疫组化对1例纵隔尤文肉瘤/外周原始神经外胚层瘤进行观察,应用反转录-聚合酶链反应(RT-PCR)检测融合基因EWS-FLI1的表达。结果肿瘤由小圆细胞、卵圆形细胞及短梭形细胞构成,巢状、片状或列兵样排列,间质显著增生,未见典型菊形团结构。免疫组化显示CK、EMA和CD99弥漫强( )。RT-PCR检测出EWS-FLIl融合基因的表达。结论EWS/pPNET与促结缔组织增生性小圆细胞肿瘤(DSRCT)具有重叠的形态学和免疫组化特点,石蜡包埋组织中检测EWS-FLI1融合基因的表达可作为诊断EWS/pPNET的可靠指标。     

A novel type of EWS-CHOP fusion gene in two cases of myxoid liposarcoma

Fusion genes consisting of TLS/FUS and CHOP or EWS and CHOP are characteristic markers for myxoid/round cell liposarcomas (MLS/RCLS). Several different structures of the fusion genes were reported in the case of the TLS/FUS-CHOP form, whereas only one type of structure has so far been found for the EWS-CHOP form, which consisted of exons 1 to 7 of the EWS and exons 2 to 4 of the CHOP gene. Here we describe a novel type of EWS-CHOP fusion gene in two cases of MLS/RCLS, which were found in a consecutive analysis of 21 cases. This fusion gene consisted of exons 1 to 10 of the EWS and exons 2 to 4 of the CHOP gene. The two cases with this fusion gene shared several clinical features, such as a large tumor mass, rapid and invasive growth, and local recurrence within 12 months after surgical resection. Histopathological findings also showed common features characterized by the diffuse proliferation of small spindle cells with a primitive mesenchymal appearance. The association of these clinical and histopathological features suggests a distinct biological property for this rare type of fusion product.     

EWS/ATF1 expression induces sarcomas from neural crest–derived cells in mice

Clear cell sarcoma (CCS) is an aggressive soft tissue malignant tumor characterized by a unique t(12;22) translocation that leads to the expression of a chimeric EWS/ATF1 fusion gene. However, little is known about the mechanisms underlying the involvement of EWS/ATF1 in CCS development. In addition, the cellular origins of CCS have not been determined. Here, we generated EWS/ATF1-inducible mice and examined the effects of EWS/ATF1 expression in adult somatic cells. We found that forced expression of EWS/ATF1 resulted in the development of EWS/ATF1-dependent sarcomas in mice. The histology of EWS/ATF1-induced sarcomas resembled that of CCS, and EWS/ATF1-induced tumor cells expressed CCS markers, including S100, SOX10, and MITF. Lineage-tracing experiments indicated that neural crest–derived cells were subject to EWS/ATF1-driven transformation. EWS/ATF1 directly induced Fos in an ERK-independent manner. Treatment of human and EWS/ATF1-induced CCS tumor cells with FOS-targeted siRNA attenuated proliferation. These findings demonstrated that FOS mediates the growth of EWS/ATF1-associated sarcomas and suggest that FOS is a potential therapeutic target in human CCS.     

ETV1、FOXF1表达与胃肠间质瘤

胃肠间质瘤(gastrointestinal stromal tumors,GISTs)是消化道最常见的间叶组织肿瘤,起源于消化道肌层Cajal细胞(interstitial cells of Cajal, ICC)或其同源干细胞,其发生多数与KIT或PDGFRA基因突变有关。GISTs的发病率、靶向治疗耐药率及术后复发率逐年升高,极大影响了患者预后,故寻找新的治疗方法已成为当前GISTs的研究热点。ETV1是转录因子ETS家族成员,可刺激KIT基因转录,KIT蛋白通过MEK-MAPK信号通路增强ETV1的表达,ETV1和KIT正反馈协同调节导致ICC/GISTs细胞内信号通路持续激活,从而促进肿瘤增殖。FOXF1在GISTs中呈特异性高表达, 可能与KIT和ETV1上游调节因子促进ICC/GISTs谱系特有基因表达有关。ETV1、FOXF1可能为GISTs的治疗提供新思路、新方向。本文就ETV1、FOXF1表达与GISTs的关系作一综述。     

EWS-Fli1 antisense oligodeoxynucleotide inhibits proliferation of human Ewing's sarcoma and primitive neuroectodermal tumor cells.

The translocation t(11;22) is a common chromosomal abnormality detected both in Ewing's sarcoma and in primitive neuroectodermal tumor cells. The translocation results in an EWS-Fli1 fusion gene, made up of the 5' half of the EWS gene on chromosome 22 fused to the 3' half of the Fli1 gene on chromosome 11. Recent studies have evaluated possible roles of the fusion gene products. However, the biological significance of EWS-Fli1 is still unknown. Using a competitive polymerase chain reaction technique, we show here that there might be a correlation between the expression levels of the EWS-Fli1 fusion gene and the proliferative activities of Ewing's sarcoma and primitive neuroectodermal tumor cells. When the EWS-Fli1 expression is inhibited by antisense oligodeoxynucleotides against the fusion RNA, the growth of the tumor cells is significantly reduced both in vitro and in vivo. The data further indicate the growth inhibition of the cells by the antisense sequence might be mediated by G0/G1 block in the cell cycle progression. These results suggest that EWS-Fli1 may play an important role in the proliferation of the tumor cells, and the EWS-Fli1 fusion RNA could be used as a target to inhibit the growth of Ewing's sarcoma and primitive neuroectodermal tumor with the specific antisense oligonucleotide.     

Bone graft gel: autologous growth factors used with autograft bone for lumbar spine fusions

A method to combine autologous growth factors (AGF) with autologous bone graft in a bone graft gel for spine fusions is described. The bone graft gel can be inserted into cages for interbody fusions or used directly for posterolateral intertransverse fusions. Sixty patients have undergone spinal fusion surgery under this technique. No equipment problems have been encountered and no adverse effects observed that could be attributed to AGF. Early clinical outcomes indicated solid or maturing fusions in 58 of 60 patients. AGFs to enhance bone healing represent an economical and readily available autologous source of growth factors.     

婴儿急性白血病的临床和分子生物学特点

目的　对 2岁以下的婴儿急性白血病作临床和分子生物学特点的研究。方法　用R带和 (或 )G带分带技术进行核型分析 ,DNA印迹法检测HRX重排 ,聚合酶链反应和逆转录 聚合酶链反应进行融合基因检测。结果　 2 0例患者经检测后其中 10例有HRX基因重排 ,该 10例作融合基因检测 ,5例是AF 4 HRX ,2例是AF 9 HRX ,1例是HRX ENL ,1例是HRX自我融合 ,1例是未曾报告过的命名为HRX EEN的融合基因。结论　婴儿急性白血病有高发的HRX基因重排 ,尤其以形成AF 4 HRX融合基因更多见 (5 0 % ) ,这对指导临床治疗和进一步的病因学研究有重要意义。     

MLL基因重排急性白血病患儿的临床和生物学特点研究

目的对儿童急性白血病进行混合谱系白血病(mixed lineage leukemia，MLL)基因重排的临床和实验研究：方法应用MLL双色荧光原位杂交(FISH)基因探针进行双色FISH，以13对引物行多重RT—PCR检测融合基冈，并联合染色体R带核型分析及流式细胞仪免疫表型检测，对298例儿童急性白血病中16例含有MLL基因重排患儿进行分析：结果MLL基因重排的白血病在儿童急性白血病中占5.4％，而在婴幼儿白血病中占56．3％；对106例患进行多重RT—PCR，检测到涉及MLL基因重排11例，其中MLL／AF42例，MLL／AF61例，MLL／AF6、MLI／ELL合并MLL／AFX或HOX11各1例，MLL／AF92例，MLL／AF101例，MLI／ELL2例。串联重复dup11q231例，HOX活化1例；27例进行了FISH检测，发现9例有MLL重排，其阳性率为36．0％；16例MLL基因重排患儿中14例(87．5％)有克隆性染色体异常，其中11例累及11号染色体[t(4；11)2例，t(6；11)、t(8；11)、t(7；8；11)、t(9；11)各1例， 112例，11q-3例]；16例MLL基因重排患儿中11例为B系ALL，以B祖细胞或前B细胞白血病为主，5例为急性单核细胞白血病，其中3例有CD7和CD2淋系抗原表达；16例MLL基因重排患儿中8例接受治疗，7例获得完全缓解。结论多重RT—PCR和FISH联合是检测MLL重排最精确和灵敏的方法，MLL基冈重排中包括易位、缺失和重复。MLL基因重排的检测对儿童急性白血病预后判断和治疗方案的选择具有重要意义，并且对WHO分型将其单独列为11q23／MLL白血病提供了有力的依据。     

Extraskeletal and skeletal myxoid chondrosarcoma: A multiparameter analysis of three cases including cytogenetic analysis and fluorescence in situ hybridization

Background: Myxoid chondrosarcoma (MCS) is a rare, low-grade, indolent tumor that can occur in soft tissue and bone. It is, however, capable of distant metastases. Previous cytogenetic data include a translocation, t(9;22)(q22-31;q12), occurring in 6 of 14 cases of the extraskeletal variant of the disease. Recently, rearrangement of the EWS gene has been reported in MCS.Methods and Results: Three cases of MCS, two skeletal and one extraskeletal, were examined to identify primary cytogenetic changes and correlate these with immunohistochemical, ultrastructural, and flow-cytometric analysis. The extraskeletal variant of MCS revealed a clonal translocation, t(9;22)(q22;q12), and trisomy for chromosomes 5, 7, 8, 12, 18, and 19. Our two cases of skeletal MCS showed complex karyotypes. In one skeletal tumor, a cryptic translocation involving chromosome 6p21.3 was identified by fluorescence in situ hybridization analysis, using chromosome-specific libraries.Conclusions: Thus far, 50% of cases of extraskeletal MCS, including our cases, have demonstrated a specific translocation, t(9;22)(q22-31;q12). Identifying this translocation is useful in confirming the diagnosis of MCS. Additional cytogenetic and molecular analysis is useful for detecting this translocation, and is also essential to determine other regions of possible diagnostic importance, such as the 6p21.3 breakpoint demonstrated in the present study. These techniques may be most useful for the skeletal lesions, in light of their heterogeneous cell populations and karyotypic variability. (Mol Diagn 1996 Jun;1(2):99-107)     

骨髓增生异常综合征患者ETV6基因重排分析及其临床意义

目的 用信号分离原位杂交（Split-signal FISH）技术分析骨髓增生异常综合征（MDS）患者ETV6基因重排情况，并初步探讨其与MDS分期和预后的关系。方法 用常规细胞遗传学和Splitsignal FISH技术分析58例MDS患者ETV6基因重排，对涉及9p24、12p13平衡易位的患者以ETV6F1、ETV6F2和JAK2R1、JAK2R2为引物进行RT—PCR。用χ^2检验和时序检验对患者的ETV6重排与MDS分期、预后之间关系进行分析。结果 检出ETV6基因重排4例（6．9％），其中1例（1．7％）经RT-PCR证实形成了ETV6／JAK2融合基因。平均随访12个月，4例（100％）重排患者全部转为急性白血病，中位生存时间7个月，另外54例患者10例（17％）转为急性白血病，中位生存时间28个月。4例重排患者均为MDS难治性贫血伴原始细胞增多（RAEB）阶段，另外54例有17例（31．5％）为MDS晚期阶段。结论 ETV6基因雷排在MDS中有较高表达率（6．9％）．并与MDS的分期和预后密切相关。