Biomedical subjects

Massimo Serra

Publications and source records attributed to Massimo Serra.

26 records · Page 2Linked to original sources

PubMed2002

Different simian virus 40 genomic regions and sequences homologous with SV40 large T antigen in DNA of human brain and bone tumors and of leukocytes from blood donors.

BACKGROUND: Many studies found only a small fragment of the large T-antigen coding sequences in human tumors, raising doubts on authenticity of SV40 sequences detected in these samples. METHODS: Five different regions of SV40 DNA were investigated in 106 fresh human tumor biopsies (25 brain, 69 bone, 12 Wilms' tumors), 71 tumor-derived cell cultures (38 from brain and 33 from bone tumors) and normal tissues (5 fresh bone biopsies and 38 buffy coats) by polymerase chain reaction (PCR) techniques and filter hybridization with specific oligoprobes. Expression of SV40 Tag sequences was analyzed in human tumor specimens by RT-PCR. RESULTS: SV40 large T-antigen sequences were detected at high prevalence, in human biopsies of primary brain (37-44%) and bone (21-37%) tumors, in cell cultures derived from brain (30-54%) and bone (53-80%) tumors. SV40 Tag sequences were detected in 29% of buffy coats of blood donors. However, only four brain tumor cell lines showed all the five regions of the SV40 genome investigated. Expression of SV40 Tag sequences was found in 11 of 27 (41%) human tumor samples. DNA sequence analysis indicated that the PCR-amplified products belong to the SV40 wild type. Polymerase chain reaction products of Tag middle portion from 20 of 78 (26%) samples showed a 97% homology with telomeric sequences of human chromosomes 10 and 11. CONCLUSIONS: Authentic SV40 sequences were detected in human samples. The expression of SV40 Tag sequences indicates that SV40 could play a role, as a cofactor, in the onset/progression of specific human cancers. The inability to detect some regions of the virus genome may suggest that those regions are not required for tumor persistence or growth and have been lost or, alternatively, may be the result of assay conditions that were unable to PCR-amplify those regions in the tumors.

Antigens, Polyomavirus Transforming↗

PubMed2002

Preferential expression of RB1-inducible coiled-coil 1 in terminal differentiated musculoskeletal cells.

RB1-inducible coiled-coil 1 (RB1CC1) is a nuclear DNA-binding protein that can induce RB1 (retinoblastoma 1) expression. RB1CC1 is abundantly expressed in human musculoskeletal and cultured osteosarcoma cells. The present study analyzed the expression of RB1CC1 and RB1 in osteosarcoma cells and in musculoskeletal cells of human embryos to evaluate the contribution of both genes to the maturational process of musculoskeletal cells. The amount of RB1CC1 message was closely related to RB1 expression in various osteosarcoma cell lines. RB1CC1 expression was difficult to detect in immature proliferating chondroblasts or myogenic cells in human embryos, but became obvious and prominent concomitantly with the maturation of osteocytes, chondrocytes, and skeletal muscle cells. RB1CC1 expression in these musculoskeletal cells increased with RB1 expression, which is linked to the terminal differentiation of many tissues and cells. In addition, the introduction of wild-type RB1CC1 decreased the formation of macroscopic colonies in the cell growth assay. Accordingly, both RB1CC1 and RB1 genes preferentially co-expressed and contributed to the maturation of human embryonic musculoskeletal cells, and may regulate the proliferative activity and maturation of tumor cells derived from these tissues.

Autophagy-Related Proteins↗

PubMed2002

The expression of ccn3(nov) gene in musculoskeletal tumors.

The CCN3(NOV) protein belongs to the CCN [cysteine-rich CYR61, connective tissue growth factor (CTGF), nephroblastoma overexpressed gene (Nov)] family of growth regulators, sharing a strikingly conserved multimodular organization but exhibiting distinctive functional features. Although previous studies have revealed an expression of CCN3 protein in several normal tissues, including kidney, nervous system, lung, muscle, and cartilage, less is known about its expression in tumors. In this study, we analyzed the expression of CCN3 in musculoskeletal tumors, using a panel of human cell lines and tissue samples. An association between CCN3 expression and tumor differentiation was observed in rhabdomyosarcoma and cartilage tumors, whereas, in Ewing's sarcoma, the expression of this protein seemed to be associated with a higher risk to develop metastases. CCN3 expression was found in 15 of 45 Ewing's sarcoma tissue samples. In particular, we did not observe any expression of CCN3 in the 15 primary tumors that did not develop metastases. In contrast, 15 of the 30 primary tumors that developed lung and/or bone metachronous metastases showed a high expression of the protein (P < 0.001, Fisher's test). Our studies indicate that CCN3 is generally expressed in the cells of the musculoskeletal system. This protein may play a role both in normal and pathological conditions. However, the regulation of CCN3 expression varies in the different neoplasms and depends on the type of cells. Thus, as reported for other CCN genes, the biological properties and regulation of expression of CCN3 are dependent on the cellular context and the nature of the cells in which it is produced. Further studies will help to clarify the biological role of this protein in musculoskeletal neoplasms.

Bone Neoplasms↗

PubMed2002

Effectiveness of insulin-like growth factor I receptor antisense strategy against Ewing's sarcoma cells.

The insulin-like growth factor I receptor (IGF-IR) plays an essential role in the establishment and maintenance of transformed phenotype of Ewing's sarcoma (ES) cells, and interference with the IGF-IR pathways by a neutralizing antibody causes reversal of the malignant potential of this neoplasm. In this paper, we stably transfected an IGF-IR antisense mRNA expression plasmid in an ES cell line to determine the effectiveness of antisense strategies against the in vitro and in vivo growth of ES cells. Doxorubicin sensitivity of TC-71 cells expressing antisense targeted to IGF-IR mRNA was also examined. Cells carrying antisense IGF-IR had a reduced expression of the receptor, a modest decrease in cell proliferation, a significant increase in anoikis-induced apoptosis, and a severely reduced ability to form colonies in soft agar. Moreover, TC/AS cells showed a marked reduction in their motility. In vivo, when cells carrying antisense IGF-IR were injected subcutaneously in nude mice, tumor formation was delayed and survival increased. Metastatic ability of ES cells was also significantly reduced. Furthermore, TC/AS clones showed a significantly higher sensitivity to doxorubicin - a major drug in the treatment of ES. These results indicate that inhibiting IGF-IR by antisense strategies may be relevant to the clinical treatment of ES patients by reducing the malignant potential of these cells and enhancing the effectiveness of chemotherapy.

Animals↗

PubMed2002

Effectiveness of Ecteinascidin-743 against drug-sensitive and -resistant bone tumor cells.

PURPOSE: The identification of new drugs is strongly needed for bone tumors.Ecteinascidin-743 (ET-743), a highly promising antitumor agent isolated from the marine tunicate Ecteinascidia turbinata, is currently under Phase II clinical investigation in Europe and the United States for treatment of soft tissue sarcoma. In this study, we analyzed the preclinical effectiveness of this drug in osteosarcoma and Ewing's sarcoma. EXPERIMENTAL DESIGN: The effects of ET-743 were evaluated against a panel of human osteosarcoma and Ewing's sarcoma cell lines characterized by different drug responsiveness and compared with the effects of standard anticancer agents. In addition, combination treatments with ET-743 and the other standard chemotherapy agents for sarcoma were analyzed to highlight the best drug-to-drug interaction RESULTS: A potent activity of ET-743 was clearly observed against both drug-sensitive and drug-resistant (multidrug-resistant, methotrexate- and cisplatin-resistant) bone tumor cells at concentrations that are easily achievable in patients (pM to nM range). Ewing's sarcoma cells appeared to be particularly sensitive to the effects of this drug. The analysis of the effects of ET-743 on cell cycle, apoptosis, and differentiation indicated that both osteosarcoma and Ewing's sarcoma cells had a slower progression through the different phases of the cell cycle after treatment with ET-743. However, the drug was able to induce a massive apoptosis in Ewing's sarcoma but not in osteosarcoma cells. In the latter neoplasm, ET-743 showed a differential effect, as indicated by the significant increase in the expression and activity of alkaline phosphatase, a marker of osteoblastic differentiation. Concurrent exposure of cells to ET-743 and other chemotherapeutic agents resulted in greater than additive interactions when doxorubicin and cisplatin were used, whereas subadditive effects were observed with methotrexate, vincristine, and actinomycin D. CONCLUSIONS: Overall, these results encourage the inclusion of this drug in the treatment of patients with bone tumors, although a careful design of new regimens is required to identify the best therapeutic conditions.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2001

Inhibition of insulin-like growth factor I receptor increases the antitumor activity of doxorubicin and vincristine against Ewing's sarcoma cells.

Innovative treatment modalities are needed for Ewing's sarcoma (ES), a neoplasm with a disappointingly low survival rate despite the use of aggressive multimodal therapeutic approaches. We and others (D. Yee et al., J. Clin. Investig., 86: 1806-1814, 1990; K. Scotlandi et al., Cancer Res., 56: 4570-4574, 1996) have previously shown the existence and the pathogenetic relevance of an autocrine loop, mediated by the insulin-like growth factor-I receptor (IGF-IR), which is crucial for survival and proliferation of ES cells in vitro. Moreover, we reported that the IGF-IR-blocking monoclonal antibody (MAb), alphaIR3, as well as suramin, a drug that can interfere with growth factor by binding to the receptors, inhibited both the tumorigenic and the metastatic ability of ES cells in athymic mice. In this study, we analyzed whether agents that can block the IGF-IR-mediated loop are of value in association with conventional cytotoxic drugs for the design of more effective therapeutic regimens. Both alphaIR3 MAb and suramin treatment significantly increased the antitumor in vitro effects of doxorubicin and vincristine, two drugs with a leader action on ES. These findings were obtained by both simultaneous and sequential treatments. Analysis of the proliferation rate and of apoptosis revealed that alphaIR3 MAb and suramin significantly enhanced the G(1)-phase rate induced by doxorubicin, without substantially affecting doxorubicin-G(2)-M-blockage of cell cycle, and significantly increased the induction of apoptosis, which confirmed that the specific blockage of IGF-IR deprives ES cells of an important tool for the prevention of drug-induced apoptosis. Moreover, combination treatments of doxorubicin plus alphaIR3 MAb significantly increase the doxorubicin-induced impairment of the ability of ES cells to form colonies in soft agar. In conclusion, we showed that, in ES, the blockage of IGF-IR by a neutralizing MAb or by suramin may greatly potentiate the antitumor activity of conventional chemotherapeutic drugs.

Antibodies, Monoclonal↗

PubMed2000

Application of CARD-ISH for Assessment of Numerical Chromosome Aberrations in Interphase Nuclei of Human Tumor Cells.

In this study based on the study of a centromeric DNA probe specific for chromosome 8 the authors standardized a method for the enzymatic detection of specific chromosome copy numbers on interphase nuclei from tumor tissue samples. Since the in situ hybridization (ISH) of chromosome 8 specific probe was revealed with a catalyzed reporter deposition (CARD), which allows a high amplification of the hybridization signal, the method was designated as CARD-ISH. This method has been standardized on interphase nuclei isolated from clinical samples of pituitary adenomas, as well as on human normal lymphocytes. On the same samples, they also evaluated chromosome 8 copy number distribution by FISH. Comparison between CARD-ISH and FISH results showed no significant differences between the two methods, proposing CARD-ISH as a reliable alternative to FISH for chromosome numerical aberration assessment in laboratories that do not have specific facilities for epifluorescence microscopy or cytogenetics and that need a long-term storage of slides that had been used for diagnostic purposes. Int J Surg Pathol 8(3):201-206, 2000

Journal Article↗

PubMedDate not supplied

Identification of markers of possible prognostic value in 57 giant cell tumors of bone.

Giant cell tumor of bone (GCT) is a neoplasm characterized by the presence of large numbers of multinucleated osteoclast-like giant cells, together with mononuclear spindle-shaped cells. Although GCT can be considered a benign lesion, it may exhibit a high biological aggressiveness, which is often associated with enhanced osteolytic properties and development of lung metastasis. By selecting different groups of GCT patients, including patients without evidence of relapse after a median follow-up of 114 months and patients who recurred with lung metastasis, this study focused on the analysis of the expression at clinical onset and in the metastasis of a series of markers involved either in bone resorption modulation or in the metastatic process. By using immunohistochemistry, we analyzed the expression of interleukin-6 (IL-6), a cytokine stimulating bone resorption that has been demonstrated to be released by GCT cells. The expression of factors of the urokinase-type plasminogen activation system, including the urokinase-type plasminogen activator (u-PA) and the plasminogen activator inhibitor type 1 (PAI-1), which have been described to be frequently implicated in the process of degradation of the extracellular matrix during the metastatic process, were also analyzed. Finally, since the action of plasminogen activators is facilitated by the presence of specific receptors on cell surfaces, the analysis included also the u-PA receptor (u-PAR). Our results showed that all these proteins were variably either expressed or overexpressed both in primary tumors and lung metastasis. However, both the level of expression and the incidence of overexpression were higher in primary GCT that relapsed and in lung metastasis compared to primary tumors from disease-free patients, suggesting a possible association of these proteins with a higher biologic aggressiveness of GCT cells. The parallel analysis of a group of primary tumors and of their respective lung metastasis demonstrated that the enhanced expression of one or more of these proteins may confer a selective advantage to GCT cells in terms of systemic invasiveness. Therefore, the evaluation of the expression levels of these proteins at the time of diagnosis may be taken into consideration for a classification of GCT into categories characterized by a different risk to relapse.

Adult↗