A cytogenetic and molecular study for improving the quality of the pathologic diagnosis of soft tissue tumors

本研究的目的是获得有助于提高软组织肿瘤诊断和鉴别诊断的细胞遗传学和分子数据。(1)利用荧光原位杂交（FISH）和比较基因组杂交（CGH）技术，证实了Bednar肿瘤的环状染色体是由17号和22号染色体的扩增物质组成的。(2)尽管皮肤纤维肉瘤隆突（DFSP）和皮肤纤维瘤（DF）之间的组织病理学区分通常是困难的，但我们的CGH分析表明，17q和22q序列的过度代表在DFSP中是常见的，而在DF中则不是。因此，CGH似乎有助于区分DFSP和DF。(3)对滑膜肉瘤石蜡包埋组织进行微束显微解剖和巢式RT-PCR检测发现，双相滑膜肉瘤上皮细胞和梭形细胞成分中均检测到SYT-SSX融合转录物，而对照组织中未检测到。我们的结果进一步证实滑膜肉瘤是单克隆性的。(4)应用CGH检测50例周围神经鞘肿瘤（PNSTs）的相对染色体拷贝数变化。在nf1相关的神经纤维瘤中，最常见的缺失发生在染色体17 [17p11.2-p13]， 9例（60%）；17q24-25 6例（40%），19例(19p13.2 8例（53%）；而在散发性神经纤维瘤和神经鞘瘤中，不到50%的病例检测到17号和19号染色体的缺失。nf1相关的MPNSTs表现为染色体17q和X的增加（各2/4例），而散发性MPNSTs表现为染色体4q的增加（3/5例）。(5)在比较基因组杂交分析的27例弹性纤维瘤中，最常见的复发增加发生在染色体位置Xq12-q22。染色体区域可能包含参与至少一些弹性纤维瘤发展的基因。(6)我们建立了一种新的人类细胞系JN-DSRCT-1，该细胞系来自于一名患有结扎性小圆细胞瘤（DSRCT）的7岁男孩。培养的细胞表现出病理特征t(11;22)（p13;p12）和Ewing肉瘤基因外显子10与Wilms肿瘤基因外显子8融合的嵌合转录信息。该细胞系将为人类DSRCT的发病机制、生物学行为和治疗模型等多种重要研究提供依据。少

The purpose of this study is to obtain cytogenetic and molecular data useful for the improvement of the diagnosis and differential diagnosis of soft tissue tumors.Results. (1) By using fluorescence in situ hybridization (FISH) and comparative genomic hybridization (CGH), we demonstrated that the ring chromosome in Bednar tumor is composed of amplified material from chromosomes 17 and 22. (2) Although histopathological differentiation between dermatofibrosarcoma protuberans (DFSP) and dermatofibroma (DF) is often difficult, our CGH analysis demonstrated that the overrepresentation of 17q and 22q sequences was a common finding in DFSP but not in DF. Thus, CGH seems to be useful for distinguishing DFSP from DF. (3) A microbeam microdissection and nested RT-PCR applied on paraffin-embedded tissue of synovial sarcomas showed that SYT-SSX fusion transcript was detected in both epithelial and spindle cell components of biphasic synovial sarcomas, but not in the control tissue. Our results con … More firm that the synovial sarcoma is of monoclonal origin. (4) CGH was used to detect changes in relative chromosome copy number in 50 cases of peripheral nerve sheath tumors (PNSTs). In NF1-associated neurofibromas, most frequent losses were found in chromosomes 17 [17p11.2-p13 in 9 cases (60%); 17q24-25 in 6 cases (40%)] and 19 [19p13.2 in 8 cases (53%); 19q13.2-qter in 8 cases (53%)], whereas in sporadic neurofibromas and schwannomas, losses of chromosomes 17 and 19 were detected in less than 50% of cases. NF1-associated MPNSTs exhibited gains of chromosomes 17q and X (2/4 cases each), whereas sporadic MPNSTs showed gains of chromosome 4q (3/5 cases). (5) In 27 elastofibromas analyzed by comparative genomic hybridization, the most common recurrent gains were found at chromosomal locations Xq12-q22. The chromosomal region possibly contain genes involved in the development of at least some elastofibromas. (6) we established a new human cell line, JN-DSRCT-1, from a 7-year-old boy with desmoplastic small round cell tumor (DSRCT). The cultured cells exhibited a pathognomonic t(11;22)(p13;p12) and a chimeric transcriptional message of the Ewing's sarcoma gene exon 10 fused to the Wilms' tumor gene exon 8. This cell line will be useful for a variety of important studies such as the pathogenic mechanism, biologic behavior, and therapeutic model of human DSRCT. Less