The Establishment And Optimization Of Molecular Diagnostic Techniques In Soft Tissue Small Round Cell Sarcoma

Objective: Establishing and optimizing molecular diagnostic techniques in soft tissue small round cell sarcomas, detected fusion genes and investigating RNA extracted methods from paraffin-embedded and formalin-fixation (PEFF)tissues by using multiplex RT-PCR.Methods: 43 cases of soft tissue small round cell tumor were collected from archives of Department of pathology in Shihezi University School of Medicine and the Department of Pathology of People′s Hospital of Xinjiang,including twenty Ewing's sarcoma/Peripheral primitive neuroectodermal tumors (ES/PNET), nine Alveolar rhabdomyosarcomas(ARMS), three myxoid liposarcomas(MLS),six embryonal rhabdomyosarcoma (ERMS), five poorly differentiated and epithelial synovial sarcoma(SS). 22 cases of control tumors,including ten malignant lymphomas(ML),one neuroblastoma,eleven biphasic and monophasic type SS. Combined PrimerPrimier5 with Oligo6.0 software to design specific multi-primers for detecting EWS-FLI,PAX3-FKHR,SYT-SSX fusion genes in one reaction system by Multiplex RT-PCR,and compare with traditional RT-PCR. At the same time 50 cases PEFF soft tissue tumor and ML were collected, compared the content and quality of total RNA with two extracted methods (AGPC and Trizol). To investigate a good methods make tissue keep the integrity of histomorphology and the quality of nucleic acid,fixed 8 cases fresh tissue of soft tissue sarcoma with 10% neutral formalin in PBS buffer solution with 6,12,24 hours respectively,and compared the integrity of histomorphology and the content of nucleic acid in different fixed time.Results: Sequencing indicated 4 Of the 5 cases of SS expressed SSX-SYT chimeric...