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目的:探讨在透明细胞肉瘤(CCS)石蜡包埋组织中检测EWS-ATF1融合基因的可行性及EWS-ATF1融合基因表达在透明细胞肉瘤诊断中的意义。方法:收集1992年～2006年期间确诊的26例CCS石蜡包埋组织标本,20例非CCS石蜡包埋组织标本为对照组,β-肌动蛋白和β2微球蛋白作为内参照,用逆转录聚合酶链反应(RT-PCR)和巢式PCR检测EWS-ATF1融合基因的表达;过氧化物酶染色(SP)方法对26例CCS石蜡组织标本进行免疫组化染色,检测S-100、HMB-45蛋白的表达。结果:26例标本中,有22例β-肌动蛋白阳性;24例β2微球蛋白阳性,其中20例EWS-ATF1融合基因阳性(16例EWS-ATF1Ⅰ型,4为EWS-ATF1Ⅲ型),余4例EWS-ATF1融合基因阴性。对照组均未检测到EWS-ATF1融合基因的表达。26例CCS石蜡组织标本中S-100及HMB-45的总阳性率分别为79.2%和70.8%。结论:在CCS石蜡包埋组织中运用RT-PCR检测EWS-ATF1融合基因是可行的,可作为辅助诊断和鉴别诊断的有力工具。
Objective: To investigate the feasibility of detecting EWS-ATF1 fusion gene in paraffin-embedded tissue of clear cell sarcoma (CCS) and the significance of EWS-ATF1 fusion gene in the diagnosis of clear cell sarcoma. Methods: Totally 26 cases of CCS paraffin-embedded tissue samples were collected from 1992 to 2006. Twenty non-CCS paraffin-embedded tissue samples were taken as control group, β-actin and β2 microglobulin as internal reference, The expression of EWS-ATF1 fusion gene was detected by polymerase chain reaction (RT-PCR) and nested PCR. Immunohistochemical staining was performed on 26 cases of CCS paraffin tissues by peroxidase staining (SP) -45 protein expression. RESULTS: Of the 26 specimens, 22 were β-actin positive and 24 β2 microglobulin was positive. EWS-ATF1 fusion gene was positive in 20 of 20 (EWS-ATF1 type 16 and EWS-ATF1 Ⅲ type 4) 4 cases of EWS-ATF1 fusion gene negative. EWS-ATF1 fusion gene expression was not detected in the control group. The total positive rates of S-100 and HMB-45 in 26 CCS paraffin tissues were 79.2% and 70.8% respectively. Conclusion: It is feasible to detect EWS-ATF1 fusion gene in CCS paraffin-embedded tissue by RT-PCR, which can be used as a powerful tool to diagnose and differentiate EWS-ATF1.