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Trail-induced apoptosis and interaction with cytotoxic agents in soft tissue sarcoma cell lines.

Five human soft tissue sarcoma (STS) cell lines (HTB-82 rhabdomyosarcoma, HTB-91 fibrosarcoma, HTB-92 liposarcoma, HTB-93 synovial sarcoma and HTB-94 chondrosarcoma) were analysed for their sensitivity to tumour necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and the function of the TRAIL apoptotic pathway in these cells. TRAIL induced significant apoptosis (>90%) in HTB-92 and HTB-93 cells, whereas no effect was observed in HTB-82, HTB-91 and HTB-94 cells. TRAIL-Receptor 1 (TRAIL-R1) was expressed in TRAIL-sensitive HTB-92 and HTB-93 cell lines, but not in TRAIL-resistant HTB-91 and HTB-94 cells. HTB-82 cells, which expressed the long (c-FLIP(L)) and short (c-FLIP(S)) splice variants of the FLICE-like inhibitory protein (FLIP), were resistant to TRAIL in spite of the presence of TRAIL-R1. TRAIL-R2,-R3,-R4 and osteoprotegerin (OPG) expression did not correlate with TRAIL sensitivity. Coincubation of TRAIL and doxorubicin led to the overexpression of TRAIL-R2 resulting in a synergistic effect of doxorubicin and TRAIL in TRAIL-sensitive cell lines and in the overcoming of TRAIL-resistance in all of the TRAIL-resistant cell lines, except HTB-91, which lacked caspase 8 expression. These data suggest that TRAIL, either as a single agent or in combination with cytotoxic agents, might represent a new treatment option for advanced STS, which constitutes a largely chemotherapy-resistant disease.

Antineoplastic Combined Chemotherapy Protocols↗

Inhibition of proliferation and induction of apoptosis in soft tissue sarcoma cells by interferon-alpha and retinoids.

Uncontrolled proliferation and a defect of apoptosis constitute crucial elements in the development and progression of tumours. Among many other biological response modifiers known to influence these mechanisms, the efficacy of retinoids and interferons in the treatment of various malignant entities is currently matter of discussion. In the present study, we have investigated the effects of 9-cis-retinoic acid (9cRA), 13-cis-retinoic acid (13cRA), all-trans-retinoic acid (tRA) and interferon-alpha on proliferation and apoptosis of human soft tissue sarcoma (STS) cell lines HTB-82 (rhabdomyosarcoma), HTB-91 (fibrosarcoma), HTB-92 (liposarcoma), HTB-93 (synovial sarcoma) and HTB-94 (chondrosarcoma) in relation to p53 genotype as well as p53 expression. HTB-91, HTB-92 and HTB-94 STS cells exhibited mutant p53, whereas wild-type p53 was found in HTB-93 STS cells, and a normal p53 status in HTB-82 STS cells, carrying a silent point mutation only. Interferon-alpha, irrespective of p53 status, inhibited the proliferation of all five cell lines dose- and time-dependently. Similarly, 9cRA, 13cRA and tRA decreased the proliferation of HTB-82 and HTB-93 STS cells, whereas the proliferation of p53-mutated HTB-91, HTB-92 and HTB-94 STS cells remained unchanged. Furthermore, only 9cRA and tRA were capable of inducing apoptosis in HTB-82 and HTB-93 STS cells, whereas HTB-91, HTB-92 and HTB-94 STS cells did not undergo apoptosis under the influence of 9cRA or tRA. Retinoic acid receptor (RAR)-alpha and RAR-beta mRNA were not detectable by Northern blot analysis in the five STS cell lines, whereas mRNA for the universal retinoic acid receptor, RAR-gamma, was expressed in all STS cell lines indicating that retinoid resistance was not associated with a lack of RAR expression. Apoptosis was not induced by interferon-alpha or 13cRA in any of the five STS cell lines tested. Our results indicate that within the panel of tested STS cell lines, inhibition of proliferation and induction of apoptosis result from different mechanisms which differ in their dependence upon the presence of intact p53.

Apoptosis↗

Intracellular localization of the radiation enhancer motexafin gadolinium using interferometric Fourier fluorescence microscopy.

Motexafin gadolinium (MGd) is a unique therapeutic agent that localizes in cancer cells and increases tumor response to ionizing radiation and certain chemotherapeutics. The in vitro intracellular localization, accumulation, and retention of MGd in murine EMT6 mammary sarcoma and Rif-1 fibrosarcoma cell lines were studied using interferometric Fourier fluorescence microscopy. MGd cellular uptake was semiquantified using its characteristic fluorescence emission band centered at 758 nm. Colocalization studies were performed using mitochondrial, endoplasmic reticulum, Golgi apparatus, nuclear, and lysosomal fluorescent organelle probes, and verified using interferometric Fourier spectroscopy. Cellular uptake was gradual and increased significantly with incubation time. MGd localized primarily within the lysosomes and endoplasmic reticulum, and to a lesser extent within the Golgi apparatus and mitochondria. Mitochondrial staining was increased in media without serum. No nuclear uptake was detected in the Rif-1 cells, but after 48 h nuclear uptake was observed in 15% of EMT6 cells. These results indicated that MGd accumulates within cytoplasmic compartments. The sustained intracellular localization of MGd may, in part, account for its unique radiation and chemotherapy enhancement properties. Interferometric Fourier fluorescence microscopy is a potentially powerful tool in delineating and verifying localization sites of therapeutic agents.

Animals↗

Soft tissue sarcoma of the vulva: A clinical study.

The purpose of this study is to report clinical aspects and treatment results of patients seen at Johns Hopkins. A search of the tumor registry of the Sidney Kimmel Comprehensive Cancer Center found 453 patients with malignancies of the vulva registered between 1977 and 1997. Patient and tumor characteristics, treatment methods, and follow-up were obtained from charts. Seven patients were identified with sarcoma of the vulva. Of these, one was removed from analysis due to histology. Three patients had leiomyosarcoma, two had fibrosarcoma, and one had epithelioid sarcoma. The mean age was 41. Mean time to diagnosis was 6 months. All but one of the tumors was located on the labia majora. Median tumor size was 3.5 cm. Surgery varied from wide local excision to radical vulvectomy with inguinal lymph node dissection. Surgical margins were microscopically negative in five of the six cases. Two patients had received adjuvant external beam radiation. One of them had a tumor greater than 5 cm and close surgical margins and the other had high-grade tumor, which recurred after previous surgery. Mean follow-up was 127.8 months. There have been no recurrences to date.

Adolescent↗

Myxomas of the head and neck.

Myxomas are benign mesenchymal tumors that occur rarely in the head and neck. When they do occur, they are prevalent in the jaws and occur less frequently in the subcutaneous tissues. We report ten new cases and review the clinical, structural, and behavioral characteristics of myxomas arising in various head and neck locations. All ages are affected. The most common initial complaint is the presence of a slow-growing mass. The sarcomas, especially liposarcomas and fibrosarcomas, rank high in the differential diagnosis of myxomas. These tumors are stubborn infiltrators. When a capsule is present, it is usually incomplete and insufficient to contain the tumor. Treatment is by wide surgical excision. The high recurrence rate previously reported is probably due to incomplete excision and is higher when enucleation or curettage are used. Long-term follow-up is mandatory since recurrences may develop several years after treatment.

Adolescent↗

Studies on adenovirus type 9-induced mammary fibroadenomas in rats and their malignant transformation.

After inoculating newborn W/Fu rats with adenovirus type 9, 27 of 27 females developed mammary fibroadenomas with a latency period of 14-25 weeks. No tumors were observed after inoculation with adenovirus type 5 or in males with the type 9 inoculation. After persistence of the tumors for 3-14 months, malignant transformation of the stroma resulted in different types of sarcoma in three rats: fibrosarcoma, round-cell liposarcoma, osteosarcoma and malignant mesenchymoma. In another animal the stroma of a fibroadenoma was highly cellular, suggesting a transition into fibrosarcoma. Malignant transformation of the epithelial component was not observed. Tumor cells contained adenovirus type 9-specific T-antigen, and rats with transplanted tumors were immunized to T-antigen. Mammary fibroadenomas without signs of malignant transformation developed in eight of nine female rats inoculated with adenovirus type 9 at an adult age. Neonatal thymectomy and total body x-irradiation neither significantly shortened the induction time of adenovirus 9-induced fibroadenomas nor increased the frequency of malignant transformation in females. One lipoma and one highly differentiated liposarcoma, however, appeared in two male rats. The results provide an example of the progression of a virus-induced benign tumor into a malignant neoplasm.

Adenofibroma↗

Fine-needle aspiration biopsy in a pediatric population. Report of 64 consecutive cases.

Fine-needle aspiration biopsy for diagnosis of mass lesions has seldom been used in pediatric patients. Sixty-four aspirations from pediatric patients have been reviewed in order to determine the adequacy of this technique in the pediatric age group. There were 38 benign diagnoses, 19 malignant diagnoses, and 7 unsatisfactory specimens. The benign diagnoses included inflammatory lesions, benign tumors, cystic lesions, and miscellaneous lesions. The malignant diagnoses included neuroblastomas, leukemias or lymphomas, small cell or poorly differentiated tumors, Wilms' tumors, rhabdomyosarcomas, a Ewing's sarcoma, and a postirradiation fibrosarcoma. Correlation of histologic and cytologic diagnoses was possible in 53 cases. The diagnostic sensitivity and specificity for the detection of malignant disease were 76% and 100%, respectively. These results suggest that fine-needle aspiration biopsy is a useful technique in the diagnosis of pediatric mass lesions.

Abdominal Neoplasms↗

Carcinosarcoma of the urinary bladder.

The clinical and pathologic features of three new and 35 previously reported carcinosarcomas of the urinary bladder were reviewed. This type of tumor is three times more common in men than women, and has occurred in patients from 33 to 82 years of age (average, 62 years), most of whom presented with hematuria of short duration. The tumors are usually large and polypoid, and on microscopic examination contain an intimate admixture of malignant epithelial and mesenchymal elements. The former may resemble transitional cell carcinoma, squamous cell carcinoma, adenocarcinoma or undifferentiated carcinoma; and the latter, fibrosarcoma, leiomyosarcoma, or undifferentiated sarcoma: skeletal muscle, osseous, and/or cartilaginous differentiation has occurred in from 25% to 50% of the tumors. Sixteen patients died as a result of their tumors; four others had metastatic disease and three were lost to follow-up. Twelve patients were alive and free of disease from 5 months to 7 years postoperatively. Most of the 12 underwent radical cystectomy, preceded in some cases by radiation therapy, but four underwent a more conservative operation.

Aged↗

Pleomorphic Liposarcoma: Comprehensive Genomic Analysis of 39 Cases With Comparison to Other Genomically Complex Sarcomas.

Pleomorphic liposarcoma (PLPS) is an aggressive high-grade sarcoma that often shows diverse morphological features and can mimic high-grade undifferentiated pleomorphic sarcoma (UPS)/spindle cell sarcoma or myxofibrosarcoma (MFS), especially when pleomorphic lipoblasts are sparse. The molecular profile of PLPS is distinct from well differentiated/dedifferentiated liposarcoma and myxoid liposarcoma. In this study, we investigate 39 cases of PLPS by comprehensive genomic profiling, occurring in 32 patients with available molecular data. Cases were reviewed and morphologic parameters-lipoblastic component, UPS-like, and MFS-like areas were estimated. The genomic findings were collected and compared to UPS and MFS groups studied using the same platform. The cohort included 15 females and 17 males, with a median age of 56.5 (range, 34-78). The lower extremity (n = 17) was the most common site involved, followed by upper extremity (n = 5) and pelvis (n = 5). UPS-like and MFS-like patterns were the most common morphologic variants, ranging from 15% to 95% and 20% to 90%, respectively. TP53 (87%) and RB1 (51%) mutations and copy number alterations were the most common alterations seen, followed by ATRX (36%). Compared to UPS and MFS, TP53 and RB1 gene alterations were significantly more common in PLPS. Conversely, CDKN2A/B deletions were infrequent in PLPS. Survival analysis showed that MYC amplification was associated with significantly shorter overall survival in PLPS. Among histologic variants, CYSLTR2 alterations were found to be highest in cases with predominantly pleomorphic lipoblasts; additionally, strong correlations were found between gene alteration frequencies of MFS and MFS-like PLPS, and between UPS and UPS-like PLPS. RB1 allele-specific copy number analysis showed loss of heterozygosity in 82% of cases. Our cohort of PLPS showed a complex molecular landscape with distinct genetic alterations, histologic correlations, and clinical outcomes, highlighting its unique position among genomically complex sarcomas and providing insights that may inform future diagnostic and therapeutic approaches.

Humans↗

Studies on a fractionated murine fibrosarcoma: proliferative potential of the separated cells.

A transplantable methylcholanthrene-induced fibrosarcoma of female BALB/c mice (the MC-2 fibrosarcoma) was dissociated by combined mechanical and enzymatic means, then fractionated by isopycnic centrifugation in linear albumin gradients. In some experiments recovered cells were both cultured in soft nutrietn agar and inoculated subcutaneously into syngeneic recipients. In these experiments a highly significant correlation was observed between subsequtnt colony number and rapid growth phase tumor size suggesting identity of clonigenic and tumorigenic cells. It was consistently found that clonigenic cells were markedly depleted from the low density extremes of the cell density distribution profiles suggesting that the low density neoplastic cells had irreversibly left the growth fraction. With increasing tumor age, sequential studies showed that both total and clonigenic cell density distribution profiles were variable, showing no obvious trend, suggesting that in the age (13-35 days) and size (2-8 g) range studied growth fraction changes had little selective effect on cells of any specific density. These results imply that a marked selective depletion of low density clonigenic cells (or selective accumulation of low density non-proliferative cells) must mainly occur during an earlier phase of tumor growth. Studies on several other murine solid tumors also showed maximal depletion of clonigenic cells from the least dense fractions, suggesting that this situation may be common.

Animals↗

A new hamster fibrosarcoma model for in vitro/in vivo evaluation of cancer chemotherapeutic agents.

A hamster fetal cell clone has been developed for in vitro chemotherapeutic studies as well as in an in vivo fibrosarcoma model system. Highly reproducible quantitative in vitro chemotherapeutic data can be obtained with this cell line within 5 days, and as few as 10(2) cells produce rapidly growing fibrosarcomas when injected subcutaneously into adult hamsters. We found using these cells in vitro that 1-beta-D-arabinofuranosylcytosine (ara-C) can antagonize the effect of 5-azacytidine (aza-C) if given simultaneously or if aza-C treatment is preceded by a 2-hr exposure to ara-c. Using the same cell line as in vivo model for chemotherapy it was also shown that ara-C and cyclocytidine significantly inhibited tumor growth. This hamster cell line may be quite useful as an in vitro/in vivo model system for the study of cancer chemotherapeutic agents.

Ancitabine↗

[The malignant fibrous histiocytoma of bone (author's transl)].

The primary malignant fibrous histiocytoma of bone (Stout) is a rare and autonomous sarcoma of the bone which has to be separated from the osseous sarcoma and the osseous fibrosarcoma. The malignant histiocytoma is morphologically characterized by the storiform pattern of interlacing spindle cell bundles and functionally by the phagocytosis of lipids, glycogen, hemosiderin and hematoidin. Contrary to the osteosarcomas, the main age of osseous histiocytomas are the fifth and sixth decade. Location of predilection are the metaphyses of the long tubular bones. The X-rays show moth-eaten spongiolysis, endosteal erosion and perforation of the cortex. Early metastases in the lungs are frequent. The primary treatment is a surgical one. Case record of a typical malignant osseous histiocytoma from the proximal metaphysis of the left femur with metastases in the lung of a female child, aged 14 years.

Adolescent↗

Increased expression of highly branched N-linked oligosaccharides terminating in N-acetylglucosamine residues in neoplastic and sclerodermal chicken fibroblasts.

Although neoplastic cells often show a shift towards the expression of larger N-linked oligosaccharides compared to their normal counterparts, little consideration has been given to the possibility that these changes might be a more general phenomenon characteristic of certain neoplastic and non-neoplastic proliferative disorders. Terminal N-acetylglucosamine (GlcNAc) cluster antigen (TGCA) is an immunoreactive epitope(s) of highly branched N-linked oligosaccharides terminating in GlcNAc residues. Here we have compared the expression of this antigen in normal, neoplastic and sclerodermal chicken fibroblasts by immunomorphological methods. TGCA was detectable in only a few, if any, fibroblasts of normal chicken skin or those cultured from chicken embryos. In contrast, the antigen appeared in 15 to 30% of chicken embryo fibroblasts transformed with avian sarcoma viruses and about 50% of neoplastic fibroblasts of both Rous sarcoma virus-induced fibrosarcomas and carcinogen-induced transplantable fibrosarcomas. Significantly, TGCA was also found in most activated fibroblasts in the skin of chickens with hereditary scleroderma. These results indicate that increased expression of highly branched N-linked oligosaccharides terminating in GlcNAc residues is characteristic of both neoplastic and sclerodermal chicken fibroblasts. Investigation of this phenomenon may thus provide insight into biochemical pathways involved in neoplastic transformation and pathogenesis of a number of non-neoplastic proliferative connective tissue disorders such as scleroderma. Moreover, changes in the expression of TGCA-positive oligosaccharides (or their modified biochemical counterparts in mammalian species) may have considerable value for diagnosis of several connective tissue diseases.

Acetylglucosamine↗

Subacute osteomyelitis presenting as a bone tumour. A review of 21 cases.

Twenty-one patients with subacute osteomyelitis who were initially considered to have bone tumours were reviewed, with an average follow up of 3 years. The clinical symptoms were not specific and laboratory investigations were normal. The radiographic findings were limited osteolysis surrounded by bone sclerosis in 14 cases, osteolysis without definite borders in 6, and onion-layer periosteal bone formation in one. The preoperative diagnoses included osteoid osteoma, osteosarcoma, chondroblastoma, Ewing's sarcoma, giant cell tumour, fibrosarcoma, eosinophilic granuloma, and bone tumour of unknown aetiology. The definitive diagnosis was made by surgical biopsy, histology and cultures which grew staphylococcus in 9 cases. The gross specimens all showed lymphocytes, plasma cells and granulation tissue with osteogenesis. All the patients recovered completely; 17 were treated with antibiotics and immobilisation, and 4 did not need an antibiotic. There was no recurrence of infection after curettage and excision of the infected tissues.

Adolescent↗