Insulin-like growth factor I expression by tumors of neuroectodermal origin with the t(11;22) chromosomal translocation. A potential autocrine growth factor.

Expression of insulin-like growth factor I (IGF-I) mRNA by some tumor cell lines of neuroectodermal origin has been described. To further explore the significance of IGF-I mRNA expression in these tumors, a more extensive analysis was performed. Most (9 of 10) neuroectodermal tumor cell lines with a t(11;22) translocation (primitive neuroectodermal tumor [PNET], Ewing's sarcoma, esthesioneuroblastoma) expressed IGF-I mRNA, whereas 0 of 15 cell lines without the translocation (PNET, neuroblastoma) expressed IGF-I. Furthermore, inasmuch as all neuroblastoma (12 of 12) cell lines examined expressed IGF-II RNA, the pattern of IGF expression could distinguish between these closely related tumors. CHP-100, a PNET cell line with the t(11;22) translocation, was shown to secrete both IGF-I protein and an IGF binding protein, IGFBP-2. This cell line also expressed the type I IGF receptor mRNA, and blockade of this receptor by a monoclonal antibody (alpha IR3) inhibited serum-free growth. These data demonstrate that IGF-I expression is a property of neuroectodermal tumors with a t(11;22) translocation and that interruption of an IGF-I autocrine loop inhibits the growth of these tumor cells.     

阴道尤文肉瘤/原始神经外胚层瘤临床病理观察

目的报道1例起源于阴道的骨骼外尤文肉瘤／原始神经外胚层瘤（ES/PNET），结合文献资料，复习其临床、病理、超微结构、免疫表型、细胞基因、鉴别诊断、治疗及预后等。方法该病例选白天津市病理会诊中心2001-06--2005-06间的1998例疑难病理标本，经常规制片，HE染色，又辅以免疫组化染色。结果阴道肿物呈带蒂息肉，由幼稚核深染小圆细胞组成，免疫组化呈神经及肌源性双向免疫表型，病理诊断阴道原发性ES/PNET。结论 ES/PNET诊断标准除原始小细胞外常向神经分化，必须具备免疫组化CD99及神经内分泌标记2项以上阳性，细胞遗传学分析有染色体易位t（11；22）（q24；q12）。     

皮肤原始神经外胚层肿瘤1例报道并文献复习

目的 报道1例罕见的皮肤原始神经外胚层肿瘤(PNET)并进行文献复习。方法 通过光镜及免疫组化对1例皮肤PNET的临床病理特征进行形态学观察及分析。结果 皮肤PNET：多发生于皮肤真皮及皮下组织内，由小圆细胞呈片状、巢状排列，核深染，有小核仁，胞质淡染或透明，核分裂象和坏死易见，可见假血管样腔隙及菊形团。CD99强( )，有一个以上神经标志物表达。结论 皮肤PNET属小圆细胞恶性肿瘤，临床上易与良性肿瘤混淆，组织形态与皮肤其他小圆细胞肿瘤相似，需经免疫组化，甚至电镜及基因检测方能明确诊断。     

两例急性髓系白血病伴t(6;21;8)(p22;q22;q22)复杂易位患者的临床与实验室研究

目的探讨2例急性髓系白血病（AML）伴t（6;21;8）（p22;q22;q22）复杂易位患者的临床及实验室特点.方法骨髓细胞经短期24 h培养后按常规方法制备染色体标本,R显带进行核型分析;双色双融合AML1/ETO探针进行丝裂间期及中期荧光原位杂交（FISH）检测AML1/ETO融合信号;逆转录-聚合酶链反应（RT-PCR）检测AML1/ETO融合基因转录本;综合分析临床特征.结果2例患者常规细胞遗传学分析显示均存在t（6;21;8）（p22;q22;q22）,间期和中期FISH证实了核型结果;RT-PCR检测到AML1/ETO融合基因转录本;尽管2例患者均诊断为AML-M2,但二者的免疫表型和治疗反应不同.结论t（6;21;8）（p22;q22;q22）是一种少见的t（8;21）（q22;q22）的复杂变异易位,还需要更多的病例以明确其临床特征和预后价值.     

原发性宫颈原始神经外胚层肿瘤3例临床病理分析

目的探讨原发性宫颈原始神经外胚层肿瘤(PNET)的临床病理特点、诊断与鉴别诊断及预后。方法回顾性分析3例宫颈PNET所有临床资料,对肿瘤组织进行HE切片、免疫组化染色,并进行随访。结果临床表现为阴道出血、下腹疼痛及宫颈肿物。镜下肿瘤主要由胞质少、分化幼稚的小圆细胞构成,细胞界限不清,细胞核圆形、深染,核仁不明显;其中1例可见Homer-Wright菊形团及神经胶质分化。免疫组化主要表现为CD99、vimentin、CD56、NSE和Syn(+)。3例均经手术治疗,其中2例行术前及术后辅助治疗。目前3例患者均存活,随访时间分别为15、11和10个月。结论宫颈PNET是一种罕见的高度恶性肿瘤,预后差。诊断较困难,需依靠组织形态和免疫组化,必要时行基因检测及电镜检查才能确诊。目前多采用手术加放、化疗综合治疗。     

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.     

Overexpression of PRAD1 gene in B-cell malignancy with t(11;14)(q13;q32) translocation]

PRAD1 (parathyroid adenoma 1) gene at chromosome 11q13 has been cloned from parathyroid adenomas as a putative oncogene, activated by translocation with the parathyroid hormone gene. 4.5 kb and 1.7 kb mRNA are transcribed and both have the same open reading frame of 885 bp encoding 34 kd protein of a cyclin gene family, cyclin D1. Recently, overexpression of PRAD1 gene has been reported to be correlated closely with the rearrangement of bcl-1 locus, particularly in centrocytic lymphoma. In our study, overexpression of PRAD1 gene was shown in five B cell lines with t(11;14)(q13;q32) including one centrocytic lymphoma line and 4 myeloma lines, when compared with other hematopoietic cell lines without translocation. One of the cell lines, SP-49, demonstrated a truncated mRNA of 3.4 kb, in addition to 1.7 kb of normal size. Southern blot analysis demonstrated a rearrangement with PRAD1 cDNA probe, suggesting that the gene is altered in this particular cell line. By cloning analysis, we confirmed that 1.8 kb deletion in 3' region of PRAD1 gene eliminating the destabilizing signal of PRAD1 mRNA, gave rise to the aberrant mRNA of 3.4 kb. These findings suggest that PRAD1 gene is most likely the candidate oncogene for bcl-1 activated by t(11; 14)(q13;q32) translocation. The gene alteration found in one cell line, SP-49, might also play an important role for deregulation of the gene.     

Prenatal diagnosis of partial trisomy 22 derived from a maternal t(11; 22) (q23; q11)

A fetus with partial trisomy 22 was detected by amniocentesis in a pregnant woman with balanced translocation, 46, XX, t(11; 22) (q23; q11). The aborted fetus had multiple congenital anomalies consisting of microcephaly, cleft lip and palate. The extra acrocentric chromosome was identified as der (22), t(11; 22) (q23; q11). The aborted fetus is compared with other trisomic cases described in literatures, and perinatal diagnosis of this case was discussed.     

Establishment and characterization of a clonal human extraskeletal Ewing's sarcoma cell line, EES1

Ewing's sarcoma, a small round cell sarcoma arising in soft tissue as well as the bone, is one of the most malignant tumors in children and young adults. Few established cell lines of extraskeletal Ewing's sarcoma (EES) have been reported, which made it difficult to examine the biological features of EES. Therefore, we have established a new clonal cell line of EES. We report its morphological characters, results of chromosomal and immunohistochemical analysis. A piece of tumor obtained from the 18-year-old female patient with EES was xenografted in a nude mouse. In vitro subcultured cells were then obtained from this xenograft. A clonal cell line was subsequently established by limiting dilution and designated EES1. EES1 cells had a doubling time of 24 hours. In the xenografted tumor, the cells expressed vimentin, CD99 (MIC2), neuron specific enolase (NSE) and cytokeratin. The original tumor cells also expressed vimentin, CD 99, and NSE, but was negative for cytokeratin. The morphological and immunohistochemical features of this cell line established, except for cytokeratin expression, were consistent with those of the primary tumor. Cytogenetic analysis of EES1 revealed chromosomal translocation of t(11; 12)(q24;ql2). The chimeric fusion of the Ewing's sarcoma gene in band 22q12 with the Friend leukemia virus integration-1 gene in band 11q24 was also demonstrated. Fluorescence in situ hybridization further confirmed the presence of translocation involving the Ewing's sarcoma gene in both the primary tumor and EES1 cells. In conclusion, we have established a human EES cell line EES1, which will provide a useful model for studying various aspects of human EES.     

慢性粒细胞白血病急变伴t(3;21)(q26;q22)染色体易位受累基因的研究

目的探讨慢性粒细胞白血病（CML）急性髓系白血病（AML）变伴t（3;21）（q26;q22）的受累基因.方法对1例CML AML变伴t（3;21）（q26;q22）患者细胞间期和中期分裂相细胞采用荧光原位杂交技术（FISH）检测AML1和bcr/abl基因重排,RT-PCR联合序列分析检测t（3;21）（q26;q22）受累基因.结果der（3）和der（21）染色体上均检测到AML1基因杂交信号,AML1-MDS1-Evi1、AML1-MDS1、AML1-EAP及Evi1基因均表达,未见AML1-Evi1融合基因表达,AML1-MDS1-Evi1基因表达水平是AML1-MDS1、AML1-EAP表达水平的1.58和1.54倍,患者Evi1基因表达水平是HEL细胞系Evi1表达水平的2.71倍.结论t（3;21）（q26;q22）导致形成AML1-MDS1-Evi1、AML1-MDS1融合基因及Evi1基因激活,这些继发的分子遗传学异常是CML急性变伴t（3;21）（q26;q22）患者急变发生的分子基础.     

伴t(16;21)(p11;q22)的恶性血液病的临床和实验分析

目的探讨伴t(16;21)(p11;q22)的恶性血液病的临床及实验室特征。方法骨髓细胞24 h培养后按常规方法制备染色体,用RHG显带技术进行细胞遗传学分析。结果 1例M2的患者其核型分析结果有t(16;21)(p11;q22)的异常,临床和血液学改变符合急性髓细胞白血病-M2a诊断,化疗后未获得完全缓解,中位生存期为6个月。结论 t(16;21)(p11;q22)是一类很独特的白血病亚型有关的易位,为少见的非随机的染色体易位,其临床预后差。     

Desmoplastic small round cell tumor with atypical immunohistochemical profile and rhabdoid-like differentiation

Desmoplastic small round cell tumor (DSRCT) is a rare, aggressive malignant neoplasm of unknown origin, and is comprised of small round cells with a characteristic desmoplastic stroma. DSRCT typically expresses epithelial, mesenchymal and neural markers simultaneously. We describe a case of DSRCT with an atypical immunohistochemical profile and rhabdoid-like tumor cells on electron microscopy. In the present case, the neoplastic cells were positive only for vimentin, desmin (cytoplasmic membranous pattern) and CD56, and negative for smooth muscle actin, synaptophysin, CD117, CD45, myogenin, CAM5.2, pancytokeratin, WT1, EMA, CD99, neurofilament, CD34 and p53. Ki67 showed a low proliferative activity. Electron microscopy showed focal rhabdoid differentiation. However, INI-1 (SNF-5/BAF47) demonstrated preservation of nuclear positivity in the neoplastic cells. Cytogenetic studies showed translocation t(11;22)(p13;q12) confirming an EWSR1-WT1 translocation characteristic for DSRCT, and t(1;15)(q11;p11.2) of unknown significance. This case is a diagnostic challenge because of atypical immunohistochemical profile and cytogenetic study is crucial in rendering the correct diagnosis.     

外周原始神经外胚叶瘤8例临床病理分析

目的 探讨外周原始神经外胚叶瘤的临床病理特征。方法 收集8例外周原始神经外胚叶瘤观察其临床病理特点并进行免疫组织化学分析。结果 所有病例在活检前均未能明确诊断，组织学检查该肿瘤具有一定的形态学特点如由大小一致的小圆形细胞组成，核圆形，点彩状，肿瘤细胞免疫组化表达Vimentin及CD90，结论 外周原始神经外胚叶瘤是一种极少见的肿瘤，临床表现缺乏特异性，早期诊断十分困难。根据其组织学改变和免疫组化染色结果，可与其他肿瘤鉴别。     

外周原始神经外胚层瘤和尤文肉瘤的临床病理与免疫组化研究

目的 探讨外周性原始神经外胚层瘤（ＰＰＮＥＴ）和尤文肉瘤（ＥＳ）的临床病理、免疫组化特点及诊断标准。方法 用ＨＥ染色、ＰＡＳ染色和免疫组化方法对１９例ＰＰＮＥＴ和ＥＳ进行观察,３例标本做电镜检查。结果 １２例ＰＰＮＥＴ,年龄２～１１岁,平均６．５岁;男８例,女４例,病变位于股骨３例,肩胛骨３例,左小腿１例,镜检：ＰＰＮＥＴ瘤细胞百列呈实性片状、分叶状、腺泡状和索条状,１２例均见Ｈｏｍｅｒ－Ｗｒｉｇｈｔ菊形团;ＥＳ瘤细胞形态与排列和ＰＰＮＥＴ相似,但胞浆透明,未见Ｈｏｍｅｒ－Ｗｒｉｇｈｔ菊形团,免疫组化：ＰＰＮＥＴ和ＥＳ均表达１２Ｅ７,每例ＰＰＮＥＴ都表达２项以上神经标志,而ＥＳ仅表达１项,结论 ＰＰＮＥＴ和ＥＳ为儿童常见的小细胞恶性肿瘤。Ｈｏｍｅｒ－Ｗｒｉｇｈｔ菊形团和神经标志至少２项阳性为诊断ＰＰＮＥＴ的条件     

The chromosome translocation (11;14)(p13;q11) associated with T cell acute leukemia. Asymmetric diversification of the translocational junctions

The t(11;14)(p13;q13) translocation associated with T cell acute lymphocytic leukemia generates two abnormal chromosomes, designated 11p+ and 14q-. To investigate the mechanism of t(11;14)(p13;q11) formation, we analyzed the translocation junctions of 11p+ and 14q- from two patients. The 11p+ junctions consisted of precise fusions of a pseudo recombination signal from chromosome 11 and the downstream recombination signal of the TCR D delta 2 gene segment from chromosome 14. In contrast, the 14q- junctions from both patients were diversified by random loss and addition of nucleotides at the translocation site. This asymmetric pattern of junctional diversification is typical of normal Ig/TCR gene rearrangement, and therefore implies that the t(11;14)(p13;q11) translocation arose due to aberrant activity of the Ig/TCR recombinase.     

二例伴有dic(9;20)(p11-13;q11)急性淋巴细胞白血病患者的临床和实验研究

目的探讨伴有dic（9;20）（p11-13;q11）的急性淋巴细胞白血病（ALL）的细胞形态学、免疫学、细胞遗传学特征和临床特点.方法骨髓细胞经直接法和24h短期培养后按常规方法制备染色体,采用R显带技术进行细胞遗传学分析.分别以9号和20号染色体着丝粒探针进行双色荧光原位杂交（FISH）检测.结果2例患者的临床和血液学改变符合ALL诊断,免疫表型分析B淋系标志阳性（CD10＋、HLA-DR＋）;染色体核型分析显示2例患者均为dic（9;20）：例1为45,XY,der（9）t（9;20）（p11;q11）,-20[20],例2为45,XX,der（9）t（9;20）（p13;q11）,t（9;22）（q34;q11）,-20[10]/46,idem,＋8[16]/47,idem,＋8,＋21[14];其中1例经双色FISH检测证实9号和20号染色体之间发生了相互易位,且形成双着丝粒染色体.结论dic（9;20）（p11-13;q11）是一种少见的重现性核型异常,可能和ALL有特殊的联系.FISH技术是检测该易位的可靠手段.     

儿童肿瘤及急性白血病中ICAM-1(CD54)的表达及在CIK细胞免疫治疗中的临床意义

本研究旨在探讨细胞间黏附分子1(ICAM-1,CD54)在初诊儿童肿瘤及急性白血病细胞上的阳性表达率,以了解其分布规律及其临床意义。采用免疫组织化学方法检测46例儿童实体瘤的病理组织切片ICAM-1的阳性率,通过流式细胞仪检测60例儿童急性白血病细胞上ICAM-1的阳性率。儿童实体瘤包括淋巴瘤10例,肝母细胞瘤3例,神经母细胞瘤6例,横纹肌肉瘤2例,尤文氏肉瘤6例,纤维肉瘤2例,原始神经外胚层肿瘤5例,肾母细胞瘤11例,骨肉瘤1例;急性白血病包括急性淋巴细胞白血病(ALL)20例,急性非淋巴细胞白血病(ANLL)40例(M1 6例、M2 7例、M3 7例、M4 15例、M5 5例)。结果表明,儿童肿瘤组中3例肝母细胞瘤ICAM-1全部阳性,而在淋巴瘤、横纹肌肉瘤、神经母细胞瘤及尤文氏肉瘤阳性率不高,纤维肉瘤、肾母细胞瘤及原始神经外胚层肿瘤患儿中未见ICAM-1表达。急性白血病组中ALL的ICAM-1阳性率为55%,ANLL的M1、M2、M3型ICAM-1的阳性率为65%,M4、M5型为50%。结论:ICAM-1在儿童肿瘤、急性白血病细胞上的表达呈一定变异性,其在肝母细胞瘤及ANLL(M1、M2和M3)上阳性率高,而在纤维肉瘤、肾母细胞瘤及原始神经外胚层肿瘤等中不表达。     

Myeloid cell differentiation arrest by miR-125b-1 in myelodysplastic syndrome and acute myeloid leukemia with the t(2;11)(p21;q23) translocation

Most chromosomal translocations in myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) involve oncogenes that are either up-regulated or form part of new chimeric genes. The t(2;11)(p21;q23) translocation has been cloned in 19 cases of MDS and AML. In addition to this, we have shown that this translocation is associated with a strong up-regulation of miR-125b (from 6- to 90-fold). In vitro experiments revealed that miR-125b was able to interfere with primary human CD34⁺ cell differentiation, and also inhibited terminal (monocytic and granulocytic) differentiation in HL60 and NB4 leukemic cell lines. Therefore, miR-125b up-regulation may represent a new mechanism of myeloid cell transformation, and myeloid neoplasms carrying the t(2;11) translocation define a new clinicopathological entity.