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PubMed2002

RNA splicing mediated by YB-1 is inhibited by TLS/CHOP in human myxoid liposarcoma cells.

Human myxoid liposarcoma contains a characteristic t(12;16) chromosomal translocation that results in fusion of the N-terminal domain of the translocated in liposarcoma (TLS) protein to the C/EBP homologous protein (CHOP). TLS possesses structural motifs that suggest it may participate in RNA processing. We demonstrate that in human myxoid liposarcoma cells, wild-type TLS binds to RNA polymerase II (Pol II) via its N-terminal domain and to the transcription and translation factor Y-box binding protein-1 (YB-1) through its C-terminal domain. The liposarcoma fusion protein TLS/CHOP retains the ability to bind RNA Pol II but lacks the ability to recruit YB-1 due to replacement of the C-terminal domain of TLS by CHOP. In an in vivo splicing assay, YB-1 promotes splicing of adenovirus EIA pre-mRNA predominantly to the 13S isoform. The oncogenic TLS/CHOP fusion protein inhibits this splicing function of YB-1 in a dominant negative manner. When considered in conjunction with studies on other sarcoma fusion proteins, these data suggest that aberrant RNA splicing may be a common feature of human sarcomas.

Adenovirus E1A Proteins↗
PubMed2005

Endoscopic approach to orbital apex lesions.

PURPOSE: To examine the role of transnasal and transantral endoscopic surgical approaches in the management of apical orbital lesions. METHODS: Retrospective case series. RESULTS: Three patients underwent combined orbital and endoscopic surgery for treatment of apical orbital lesions. In two of these patients, the orbital apex was decompressed, which resulted in visual recovery. Using endoscopic approaches to the posterior orbit and orbital apex allowed better visualization, with minimal manipulation of tissues in an area where surgical access is limited. CONCLUSIONS: Apical orbital lesion management benefits from a multidisciplinary and individualized plan. Endoscopic approaches can be used to improve visualization and access to a difficult surgical area. They also allow extra instrumentation to be used through the relatively capacious paranasal sinuses. These advantages may obviate lateral orbitotomy in some situations. Although endoscopic techniques are useful for biopsy, debulking, and removal of orbital apex lesions, they may be more difficult to apply to an intraconal lesion at the orbital apex.

Biopsy↗
PubMedDate not supplied

[Primary retroperitoneal tumors. Our experience].

Primary retroperitoneal tumors are rare (0.05-0.2% of all tumors), often malignant and characterized by a poor and non-specific symptomatology and by a late diagnosis. Complete resection is possible in only a few patients, while recurrence is very common. The records of 29 adult patients who underwent operative treatment at Surgical Oncology Institute-University of Cagliari between November 1973 and June 1992 were reviewed; 9 were males, 20 females, median age 46.4 years (range 12-82). There were 4 benign tumors (13.8%) and 25 malignant (86.2%). Fibrosarcoma (9 cases, 31%) and liposarcoma (3 cases, 10.3%) were the most frequent histologic types. There were also two fibroleiomyomas, leiomyosarcomas, malignant fibrous histiocytomas and neuroblastomas, one case of fibroma, neurofibroma, rhabdomyosarcoma and schwannoma. Five sarcoma were not otherwise specified. Abdominal mass (25 cases, 86.2%), flank or abdominal pain (15 cases, 51.7%) and weight loss (8 cases, 27.6%) were most common symptoms; change in bowel habit and constipation (6 cases, 20.7%), fever (5 cases, 17.2%), urinary disorders (4 cases, 13.8%) nausea and vomiting (2 cases, 6.7%) were less common. Diagnosis was made by ultrasonography, computed tomography and traditional radiographic studies. median interval between first symptoms and diagnosis was 11 months. Complete surgical resection was possible in only 13 cases (46.4%): 10 of the 25 malignant tumors (40%) and 3 of the 4 benign tumors (75%).(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMedDate not supplied

Amplification of the CDK4 gene in sarcomas: tumor specificity and relationship with the RB gene mutation.

Amplification of the CDK4 gene, which encodes a key molecule in the cell cycle, has been shown in some types of human neoplasms, including bone and soft tissue tumors. It is also reported that the CDK4 gene is coamplified with other sequences in the 12q13-15 region, including the MDM2 and SAS genes. Using 146 DNA samples derived from a variety of bone and soft tissue tumors, we have studied the pattern of amplification of these three genes, CDK4, MDM2, and SAS, to investigate whether there are any tumor type specific patterns of amplification. Amplification of at least one of these three genes was found in 18 tumors, and five different patterns of amplification were observed. Amplification of all of these three genes was detected in 9 cases. Amplification of the CDK4 gene without MDM2 amplification was observed in osteosarcomas and a chondrosarcoma but not in soft tissue tumors, whereas amplification of MDM2 gene alone was observed in malignant fibrous histiocytomas (MFHs), liposarcomas, and lipomas, but not in bone tumors. These results suggested that the CDK4 region is the primary target for amplification in bone tumors, whereas the MDM2 region is in soft tissue tumors. We also investigated the relationship of CDK4 amplification with retinoblastoma (RB) gene mutations in osteosarcomas, for which we have already performed the mutation analyses in detail. Interestingly, contrary to the prevailing theory that CDK4 amplification is an alternative mechanism for RB gene mutation, we found that three of four cases with amplification of the CDK4 gene showed loss of expression of the RB protein, one of which was proved to have an gross DNA alteration in the RB locus. This redundancy of mutations may indicate that the amplification of CDK4 may have some roles other than the inactivation of the RB protein in the development of osteosarcomas.

Bone Neoplasms↗
PubMed2003

Inconspicuous insertion 22;12 in myxoid/round cell liposarcoma accompanied by the secondary structural abnormality der(16)t(1;16).

In myxoid/round cell liposarcoma, the t(12;16)(q13;p11) and its associated fusion transcript, FUS-CHOP, characterize greater than 95% of cases. The variant translocation t(12;22)(q13;q12) and associated EWS-CHOP fusion transcript are rare. A second non-random aberration observed in roughly 20% of Ewing's sarcomas, and to a lesser extent other select sarcomas, is the unbalanced 1;16 translocation. Recognition of this secondary aberration in the absence of an obvious primary karyotypic abnormality strongly suggests that the use of other genetic approaches will be informative in uncovering a clinically suspected primary anomaly. The following case illustrates the utility of molecular cytogenetic and reverse transcriptase-polymerase chain reaction techniques in diagnosing an ins(22;12)(q12;q13q14) and associated EWS-CHOP fusion transcript in a myxoid/round cell liposarcoma exhibiting a der(16)t(1;16)(q11;q11).

Adult↗
PubMed1983

Immunoregulatory factors derived from human tumors. II. Partial purification and further immunobiochemical characterization of a human sarcoma-derived immunosuppressive factor expressing HLA-DR and immunoglobulin-related determinants.

An immunoregulatory factor (IRF) that suppresses Con A-mediated peripheral blood mononuclear cell (PBM) proliferative responses was partially purified by DEAE anion exchange chromatography and affinity chromatography from a 3 M KCl extract of a human liposarcoma. The factor (m.w. = 70K) co-purified with albumin, monitored by two-dimensional gel electrophoresis, and demonstrated a heterogeneous isoelectric point (pI 7.6-7.8). Xenoantisera produced against the DEAE-purified fraction and coupled to Affigel 10 removed suppressive activity that could subsequently be eluted by glycine-HCl, pH 3.5. An anti-albumin column partially removed activity, but the unbound 70K factor could still be detected in the column effluent. A xenoantiserum to this 70K effluent coupled to acrylamide beads completely removed the immunosuppressive activity in immunodepletion experiments. Further direct binding enzyme-linked immunoassays (ELISA) and solid-phase immunoabsorption experiments with monoclonal antibodies to human anti-HLA-DR framework determinants and a constant region of the IgM mu-chain demonstrated determinants on the 70K factor recognized by these antibodies.

Cells, Cultured↗
PubMed1994

Fusion of the FUS gene with ERG in acute myeloid leukemia with t(16;21)(p11;q22).

It has been shown that the gene ERG in 21q22 is rearranged in the t(16;21)(p11;q22) associated with acute myeloid leukemia (AML). ERG is a member of the ETS gene family and is fused with EWS in a subset of Ewing's sarcomas. EWS in 22q12 has a very high homology with FUS (also called TLS) in 16p11; the latter gene is rearranged in the t(12;16)(q13;p11) that characterizes myxoid liposarcoma. To investigate whether FUS is involved in the t(16;21) of AML, we used the Southern blot technique and polymerase chain reaction (PCR) to examine the bone marrow of a 3-year-old boy with a t(16;21)(p11;q22)-positive AML. Hybridization of Southern blot filters containing digested DNA with probes for FUS and ERG showed both germline and aberrant fragments. Using specific primers for the 5' part of FUS and the 3' part of ERG, we amplified a 4.4 kb genomic FUS/ERG DNA fragment from the leukemic sample. In a second PCR experiment, in which we used primers upstream of the 5' part of ERG and downstream of the 3' part of FUS, a 5.6 kb fragment was amplified. Blotting and hybridization with specific probes for FUS and ERG revealed that the amplified fragments consisted of FUS/ERG and ERG/FUS hybrid DNA. Both PCR fragments, when used as probes, detected germline ERG and FUS as well as aberrant fragments on Southern blot filters. The results suggest that the t(16;21) in AML leads to rearrangement and fusion of the FUS and ERG genes.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗
PubMed2000

Genetic characterization of angiomatoid fibrous histiocytoma identifies fusion of the FUS and ATF-1 genes induced by a chromosomal translocation involving bands 12q13 and 16p11.

This case report documents the first karyotypic, fluorescence in situ hybridization, and genetic analysis of an angiomatoid fibrous histiocytoma that arose and recurred in the arm of a 5.5-year-old girl. Complex rearrangements between chromosomes 2, 12, 16, and 17 were noted, as well as deletion in the long arm of chromosome 11. Flow cytometry revealed a normal cell population. The t(12;16) site was further investigated using reverse transcriptase-polymerase chain reaction. We found that the FUS (also known as TLS) gene from 16p11 combined with the ATF-1 gene from 12q13 to generate a chimeric FUS/ATF-1. The FUS gene is rearranged in the t(12;16)(q13;p11) that characterizes myxoid liposarcoma and in acute myeloid leukemia with t(16;21)(p11;q22), while the ATF-1 gene is rearranged in the t(12;22)(q13;q12) found recurrently in clear cell sarcomas (malignant melanoma of soft parts). Thus, the FUS/ATF-1 gene in angiomatoid fibrous histiocytoma is predicted to code for a protein that is very similar to the chimeric EWS/ATF-1 found in clear cell sarcoma.

Activating Transcription Factor 1↗
PubMed1988

Monoclonal antibodies that detect different antigenic determinants of the same human osteosarcoma-associated antigen.

Monoclonal antibodies (MoAbs) against human osteosarcoma cells were obtained by the fusion of NS/1 mouse myeloma cells with spleen cells from the human osteosarcoma cell line-immunized BALB/c mice. Two hybrid clones were established and designated as 2H10 and 2D3. Both MoAbs reacted strongly with all osteosarcoma tissues but not with other bone and soft tissue tumors such as chondrosarcoma, malignant fibrous histiocytoma, liposarcoma, leiomyosarcoma, and rhabdomyosarcoma. In addition, neither MoAb reacted with tumor cell lines and tissues obtained from other cancers. Immunohistochemical analysis demonstrated that 2H10 and 2D3 reacted with endothelial cells in sarcoma tissues, but not with those of other tumors and normal tissues. 2H10 also reacted with cells on the basal layer of epidermis of the skin. 2H10- and 2D3-defined antigen has an approximate molecular weight of 75,000 under nonreducing and reducing conditions, indicating that the antigen has a single chain structure and there is no intramolecular disulfide bond. 2H10- and 2D3-defined antigen has a pI value between 5.5 and 6.2. Sequential immunoprecipitation analysis clearly demonstrated that 2H10 and 2D3 recognized the same antigen molecule. However, further analysis suggested the possibility that 2H10 and 2D3 MoAbs recognized the different antigenic determinants on the same antigen molecule.

Animals↗
PubMed2000

CD34 immunohistochemistry in female genital tract carcinosarcoma (malignant mixed müllerian tumors) supports a dominant role of the carcinomatous component.

Female genital carcinosarcomas (FGTCSs) are aggressive biphasic malignant neoplasms with histologic features of both carcinomas and sarcomas. However, their behavior is dominated by the carcinomatous component. CD34 is useful in distinguishing carcinomas with sarcomatoid features from epithelioid sarcomas, as it is never expressed in carcinomas although often expressed in epithelioid sarcomas. This investigation hypothesizes that CD34 expression in FGTCSs will contribute useful histogenetic and clinical information. Paraffin sections from a surgical series of 43 FGTCSs were stained using anti-CD34 with an automated immunohistochemical stainer, antigen retrieval, and appropriate external/internal controls. Reactions were graded as follows: negative (N) or S1-3, strong reactivity in <10% = S1, 10-50% = S2, or >50% = S3 of cells, respectively. The series included 31 endometrial, 6 ovarian, 5 cervical, and 1 fallopian tubal FGTCSs, 70% exhibiting heterologous elements (22 rhabdomyosarcomas, 11 chondrosarcomas, 1 osteosarcoma, and 1 liposarcoma). The carcinomatous component included 19 endometrioid, 12 serous, 6 adenosquamous, 4 clear cell, and 2 poorly differentiated carcinomas: modified International Federation of Gynecologic Oncology grade I, 9%; II, 21%; and III, 70%. CD34 reactivity was 100% N in the carcinomatous components. CD 34 stained 21 % of sarcomatous components as follows: 2S1, 6S2, and 1S3 (three homologous and six heterologous). Of the heterologous components, only rhabdomyosarcoma stained with the following patterns: 1S1, 2S2, 1S3. Two of six heterologous FGTCSs exhibited staining of only the homologous sarcomatous component with the following pattern: 2S2, as the heterologous sarcomatous component (pure rhabdomyosarcoma in both cases) showed no staining. The staining pattern was unrelated to primary site of the FGTCSs. The pattern of CD34 expression (N in the carcinomatous component like carcinomas) supports the generally accepted opinion of dominance of FGTCS by the carcinomatous component. The staining of the sarcomatous component was rare (21%) and focal (only one of nine was S3). CD34 staining may help distinguish FGTCSs from epithelioid sarcomas, which strongly express CD34.

Aged↗