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F Basolo

Publications and source records attributed to F Basolo.

At least 91 records · Page 5Linked to original sources
PubMed1994

Expression of p21 ras protein as a prognostic factor in papillary thyroid cancer.

We studied the expression of p21, the ras encoded protein, in primary tumour of 45 patients with papillary thyroid cancer (PTC). Patients were grouped according to outcome so that one group (31 patients) had a good outcome and the other (14 patients) a fatal outcome, after a follow-up of at least 5 years. The presence of p21 ras protein was assessed by immunohistochemistry with a specific monoclonal antibody (MAb Y-13259). The results were correlated with the outcome, with the expression of proliferating cell nuclear antigen (PCNA)/cyclin (as a marker of cell proliferation) and with other well established prognostic factors for PTC (age, grading, extension and tumour size; Endocrinol Metab Clin North Am 1990, 19, 545-576). p21 staining in tumours of living patients was negative in 15, weakly positive (1+) in 10 and strongly positive (2+ or more) in 6 patients. In tumours from deceased patients, p21 staining was negative in 1, weakly positive in 2 and strongly positive in the remaining 11 patients (P < 0.001, chi 2). PCNA immunostaining was increased in 63.6% (7/11) of the tumours from deceased patients compared to 17.8% (5/28) of the tumours of living patients, but no direct correlation was found between p21 and PCNA expression. Among the other prognostic factors studied, only age > or = 40 years was a significant predictor of poor outcome. The survival curve of patients with strongly positive p21 staining was similar to that of patients aged > or = 40 years at the time of diagnosis. The combination of p21 > or = 2+ and age > or = 40 was superior to age alone (P < 0.05) as a prognostic indicator of poor outcome. In conclusion, our results indicate that the p21 product of the ras (proto)oncogene is differently expressed in PTC, in relation to the degree of aggressiveness. Regardless of the pathogenetic role of the ras oncogene in thyroid tumorigenesis, our data indicate that the expression of the p21 ras protein may be regarded as a prognostic indicator in PTC. Furthermore, overexpression of p21 ras protein is associated with patients in the older age groups, and might contribute to the poor prognosis of elderly patients.

Adolescent↗
PubMed1994

Inhibition of CRIPTO expression and tumorigenicity in human colon cancer cells by antisense RNA and oligodeoxynucleotides.

CRIPTO is an epidermal growth factor-related gene expressed in a majority of human colorectal tumors. To assess the role of CRIPTO in the growth control of human colon cancer, we have treated human colon carcinoma GEO and CBS cells, that possess high levels of CRIPTO, and WIDR colon cancer cells, that are negative for CRIPTO expression, with two antisense phosphorothioate oligodeoxynucleotides complementary to the 5' end of the human CRIPTO mRNA. Both antisense oligodeoxynucleotides significantly reduced endogenous CRIPTO protein levels and inhibited GEO and CBS cell growth in monolayer and in semisolid medium, whereas they did not affect WIDR cell growth. In addition, GEO, CBS and WIDR cells were infected with a recombinant retroviral vector containing the hygromycin-resistance gene and a 900 bp EcoRI-EcoRI coding fragment of the human CRIPTO cDNA oriented in the 3' to 5' direction. GEO and CBS CRIPTO antisense infectants exhibited a 60 to 70% reduction in CRIPTO protein expression, in monolayer growth and in soft agar cloning efficiency as compared to parental noninfected cells. In contrast, infection of WIDR cells with the CRIPTO antisense retrovirus did not alter their growth. Finally, GEO CRIPTO antisense infectants formed tumors in nude mice that were significantly smaller and had a larger latency period as compared to noninfected GEO cells.

Animals↗
PubMed1993

Normal breast epithelial cells produce interleukins 6 and 8 together with tumor-necrosis factor: defective IL6 expression in mammary carcinoma.

Virtually pure primary cultures of normal mammary epithelial cells (MEC) obtained from healthy women were shown to release interleukin 6 and 8 (IL6, IL8) and to produce a nonsecreted form of tumor-necrosis factor (TNF). No interferon (IFN), whether alpha, beta, or gamma, or IL1-alpha or -beta could be detected. Analysis of cellular RNA confirmed these findings and showed that MEC also express IL6 receptor and TNF-alpha-related mRNAs. Epithelial cells were selectively stained by antibodies to IL6, IL8 and TNF-alpha both in primary cultures and in the normal mammary gland. Samples of human milk contained sizable amounts of IL6, IL8 and IFN-gamma; yet the liquid phase was consistently negative for other cytokines (i.e., TNF-alpha, IFN-alpha/-beta, IL1-alpha/-beta). Expression of IL6 (but not of IL8 and TNF-alpha) was abolished in ductal infiltrating carcinomas and greatly reduced in cultures of oncogene-transfected mammary cells, suggesting that alterations of IL6 expression are associated with pathogenesis in breast cancer.

Adolescent↗
PubMed1993

Growth-stimulating activity of interleukin 6 on human mammary epithelial cells transfected with the int-2 gene.

We have shown recently that normal human mammary epithelial cells do produce interleukin 6 (IL6), interleukin 8, and a nonsecreted form of tumor necrosis factor. Here we report that ductal infiltrating mammary carcinomas fail to express immunoreactive IL6. Since abnormalities of cytokine genes are a frequent event in cancer, we investigated the production of and the response to cytokines of mammary cells using a panel of oncogene-transformed cells derived from the spontaneously immortalized MCF-10A cell line. We found that only the parental line and the int-2-transformed cells responded to exogenous IL6 and/or were suppressed by IL6-neutralizing antibody. In contrast to highly transformed cells, these two lines, which were either nontransformed (MCF-10A) or weakly transformed (int-2), were found to express IL6 receptors. These data suggest that loss of IL6 pathways can be a marker of mammary cell transformation.

Animals↗
PubMed1992

Matrigel promotes retinoblastoma cell growth in vitro and in vivo.

Cells derived from retinoblastomas grow slowly in vitro and only very rarely form tumors in nude mice. Matrigel, a mixture of components normally found in basement membranes, promotes the growth of Y-79 and WERI-Rb1 retinoblastoma (Rb) cells when added to suspension cultures of the 2 Rb cell lines. It also substantially increases cell adhesion in vitro. Y-79 cells, seeded into a Matrigel matrix, form round colonies over a 3-week period similar to those of control, weakly metastatic murine melanoma cells. In vivo, s.c. co-injection of Matrigel with either Y-79 or WERI-Rb 1 cells into nude mice promotes retinoblastoma tumor formation. Transplantation of as few as 1,000 cells allows for xenografting under these conditions, while no tumors were observed in the absence of Matrigel, even at 10 x 10(6) cells/inoculum. The tumors produced have the expected morphology and express an mRNA for a highly specific retina/retinoblastoma marker protein, the interphotoreceptor retinoid-binding protein. Thus, the xenografts obtained maintain the original morphological and molecular characteristics of the injected cells and represent a useful model for in vivo studies of retinoblastoma growth and treatment.

Animals↗
PubMed1992

Interferon-alpha/beta in virus-induced mouse mammary carcinogenesis: effects on the spontaneous process and on the progression of transplanted pre-neoplastic lesions.

Low levels of anti-viral activity, mainly interferon alpha/beta (IFN-alpha/beta), are regularly found in lymphoid tissues of BALB/c mice infected with the C3H strain of mammary tumor virus. At the time of tumor development, significant amounts of anti-viral activity were detected in homogenates of spleen and mammary tumors, but not in blood and normal mammary glands. This activity is pH2-resistant and neutralized by antibody to IFN/alpha-beta. The pathogenetic role of IFN in mammary carcinogenesis was investigated in 2 ways: (a) by treating virus-infected newborn mice with antibody to IFN-alpha/beta, and (b) by giving either the latter antibody or IFN-alpha/beta to virus-free animals transplanted with pre-neoplastic lesions. Mice were treated only for 2 months, starting either 1 week after birth or immediately after tumor transplant. In case (a), treatment with antibody to IFN-alpha/beta shortened the incubation period of mammary carcinomas and decreased the mean survival time. In case (b), anti-IFN antibody did not significantly affect the development of mammary tumors. However, exogenous IFN-alpha/beta markedly reduced both tumor incidence and mortality rate. These results indicate that endogenous IFN-alpha/beta plays a crucial role in the in vivo restriction of the early infectious phase of spontaneous carcinogenesis and that relatively high doses of IFN-alpha/beta may inhibit the progression of pre-neoplastic lesions.

Aging↗
PubMed1992

Regulation of surface-differentiation molecules by epidermal growth factor, transforming growth factor alpha, and hydrocortisone in human mammary epithelial cells transformed by an activated c-Ha-ras proto-oncogene.

Spontaneously immortalized human mammary epithelial cells MCF-10A were transfected with an activated c-Ha-ras oncogene. Transfected cells (MCF-10T) acquire a malignant phenotype, as already reported. Studies of 125I-2'-deoxyuridine incorporation in cultures given graded doses of hydrocortisone (HC), cholera toxin (CT), epidermal growth factor (EGF), and transforming growth factor alpha (TGF-alpha) showed that though MCF-10T had become almost independent on exogenous EGF and TGF-alpha, they continued to respond to the synergistic effect of HC and CT plus EGF. Both lines were phenotypically characterized with an immunoradiometric assay in live cells. Expression of MHC class-I molecules, human milk-fat-globule-I antigen, and EGF receptor was reduced in ras-transfected cells, although other differentiation markers were unchanged. Exogenous EGF down-regulated the expression of functional EGF-R, selectively in transformed cells. TGF-alpha failed to modulate EGF-R. In contrast, HC strongly stimulated the expression of EGF-R while depressing MHC class-I molecules. Thus, it appears that in vivo HC may co-operate with TGF-alpha and EGF in promoting the growth of transformed mammary cells. This hormone might also favor the escape from immune surveillance by reducing the expression of surface differentiation markers.

Antigens, Differentiation↗
PubMed1992

Additive effects of c-erbB-2, c-Ha-ras, and transforming growth factor-alpha genes on in vitro transformation of human mammary epithelial cells.

MCF-10A cells are a spontaneously immortalized untransformed human mammary epithelial cell line. We have previously shown that overexpression of a human point-mutated c-Ha-ras proto-oncogene, the rat c-neu (c-erbB-2) proto-oncogene, or the human transforming growth factor-alpha (TGF-alpha) gene in MCF-10A cells leads to in vitro transformation of such cells. To ascertain whether the introduction of two of these genes into MCF-10A human mammary epithelial cells induces a completely tumorigenic phenotype, we infected MCF-10A Ha-ras and MCF-10A TGF-alpha cells with a recombinant retroviral vector containing the human c-erbB-2 proto-oncogene and the hygromycin-resistance gene. Ten MCF-10A TGF-alpha/c-erbB-2 (MCF-10A TE) and 10 MCF-10A Ha-ras/c-erbB-2 (MCF-10A HE) hygromycin-resistant clones were randomly selected and expanded into cell lines. MCF-10A TE and MCF-10A HE clones expressed a 10-fold to 40-fold increase in p185 erbB-2 protein levels compared with parental uninfected cells. These cells exhibited a fourfold increase in their growth rate in serum-free medium and showed a strongly reduced mitogenic response to exogenous epidermal growth factor or TGF-alpha compared with MCF-10A cells. Moreover, both MCF-10A TE and MCF-10A HE clones exhibited a fivefold to 20-fold higher cloning efficiency in soft agar than MCF-10A Ha-ras, MCF-10A c-erbB-2, or MCF-10A TGF-alpha clones. However, neither MCF-10A TE nor MCF-10A HE cells were able to grow as tumors in vivo when they were injected into nude mice. These results suggest that c-Ha-ras, c-erbB-2, and TGF-alpha genes have an additive effect on the in vitro transformation of an immortalized human mammary epithelial cell line, but that additional genetic changes such as activation of other proto-oncogenes or inactivation of a tumor suppressor gene may be necessary to elicit a fully tumorigenic phenotype.

Animals↗
PubMed1992

Mutated c-Ha-ras oncogene alters cytokeratin expression in the human breast epithelial cell line MCF-10A.

MCF-10A, a spontaneously immortalized human breast epithelial cell line, has been transformed by transfection with the mutated c-Ha-ras oncogene obtained from T24 carcinoma cells. The pattern of cytokeratin expression was studied in MCF-10A cells in comparison with plasmid transfected or MCF-10Aneo cells, normal ras proto oncogene transfected or MCF-10AneoN cells, and transformed or MCF-10AneoT cells. Cytokeratin expression was studied by western immunoblot of total cellular proteins separated by two-dimensional gel electrophoresis. Blots with cytokeratin specific AE1 and AE3 antibodies showed identical molecular weight species of cytokeratins in MCF-10A, MCF-10Aneo, MCF-10AneoN, and MCF-10AneoT cells; however, in MCF-10AneoT cells, the intensity of immunostaining and the number of immunoreactive phosphorylated polypeptides keratins 7, 8, 15, and 16 was decreased. It was concluded that c-Ha-ras oncogene-induced transformation alters quantitatively the cytokeratin pattern of human breast epithelial cells and that these changes could explain some of the morphologic alterations observed in c-Ha-ras transformed cells.

Blotting, Western↗
PubMed1991

Transformation of human breast epithelial cells by c-Ha-ras oncogene.

To determine whether the c-Ha-ras oncogene plays a role in the initiation of mammary carcinogenesis, an immortalized human breast epithelial cell line, MCF-10A, was transfected with the plasmid vector pHo6T1 containing the T24 Ha-ras oncogene and the aminoglycoside phosphotransferase gene, which confers resistance to geneticin. Transfected cells exhibited an altered pattern of growth and tridimensional morphology in collagen gel. They also exhibited anchorage-independent growth and loss of requirement for hormones and epidermal growth factor; in addition, they expressed invasiveness and increased collagenolytic activity in an in vitro system and became tumorigenic in irradiated nude mice, all properties indicative of malignant transformation. Transformed cells contained the mutated c-Ha-ras oncogene and expressed the p21 mutated protein. These data indicate that the c-Ha-ras oncogene is capable of inducing malignant phenotypes in immortalized human breast epithelial cells.

Breast↗
PubMed1991

Cell surface glycosylation changes accompanying immortalization and transformation of normal human mammary epithelial cells.

Fluorescent lectin binding to cell surfaces was quantitatively analysed by flow cytometry on mortal human breast epithelial cells MCF-10M, the immortalized cell line MCF-10A derived from MCF-10M and sublines of MCF-10A transfected with the neomycin resistance gene (MCF-10Aneo), the c-Ha-ras protooncogene (MCF-10AneoN), or transfected and transformed with the c-Ha-ras activated oncogene (MCF-10AneoT). Immortal MCF-10A cells bound 10-fold more peanut agglutinin (PNA) and soy bean agglutinin (SBA) than did MCF-10M cells. Transformed MCF-10AneoT cells bound approximately ten times more PNA than did non-transformed cells transfected with protooncogene (MCF-10AneoN). Treatment of the transfectants with neuraminidase abrogated the differences in PNA-binding and reduced the differences in SBA binding. SDS-PAGE separation of PNA binding glycoproteins revealed different patterns for all MCF-10A derived sublines.

Blotting, Western↗
PubMed1991

Medullary thyroid cancer. An immunohistochemical and humoral study using six separate antigens.

The authors investigated the humoral and tissue expression of six antigens associated with medullary thyroid cancer (MTC): calcitonin (CT), calcitonin gene-related peptide (CGRP), carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), somatostatin (SRIF), and thyroglobulin (TG). The antigens were studied in the neoplastic C cells using immunohistochemistry with specific antisera and in the plasma using specific radioimmunoassay. Eighteen patients (8 male and 10 female patients, aged 12-72 years) were studied. Mean follow-up was 70.7 months (range, 2-179 months). Nine patients (50%) died of their disease after a mean follow-up of 47.2 months (range, 2-116 months). By immunostaining, primary tumors expressed CT and CEA in all cases and NSE was positive in 90%, CGRP in 66%, SRIF in 63%, and TG in 58%. Metastatic tissues were positive in all cases of CT staining, 92.8% of CEA, 71.4% of NSE, 73.3% of CGRP, 38.5% of SRIF, and only 13.3% of TG staining. In positive cases the percentage of positive cells and the degree of staining were variable among the different antigens. The expression of an antigen in the neoplastic cells was associated with the hypersecretion of the corresponding antigen in the circulation in the case of CT and CEA. The levels of these antigens were elevated in all patients with metastases and could accurately predict the appearance of new metastases or indicate the effective treatment of previous metastases by surgery. In the case of NSE, CGRP, and SRIF, few patients had increased plasma concentrations of the antigens and these usually occurred during very advanced phases of the disease. Detectable levels of serum TG were never observed. When the outcome of the disease was compared with the expression of CT, CEA, NSE, CGRP, and TG, no correlation could be found. On the contrary, SRIF expression in the primary tumor could differentiate two groups of patients with different survival rates. SRIF-positive patients had survival rates of 100% and 50% at five and seven years, respectively, whereas SRIF-negative patients had survival rates of 40% at five years and 25% at seven years.

Adolescent↗
PubMed1991

Surgery plus adjuvant chemotherapy for T1-3N0M0 small-cell lung cancer. Rationale for current approach.

Between January 1980 and December 1986, 42 patients with histological confirmed small-cell lung cancer (SCLC) staged pathologically as T1-3N0M0 disease entered a prospective study of surgery plus adjuvant chemotherapy including six courses of i.v. cyclophosphamide (1 g/m2, day 1), epirubicin (60 mg/m2, day 1), and etoposide (120 mg/m2, days 1, 3, and 5), at 3-week intervals. No thoracic or prophylactic cranial irradiation was given. With a 52.5-month median follow-up, estimated 5-year survival was 36% and median survival was 32.7 months (range, 5-100+) for all patients. Seven patients (17%) are currently alive and disease-free at 63 to 100 months. In univariate analysis, baseline characteristics significantly influencing survival and disease-free interval (DFI) were the tumor stage (T1 and T2 versus T3), tumor histology (oat and intermediate versus combined cell type), and type of resection (lobectomy versus pneumonectomy). However, in multivariate analysis, only the tumor stage (T1-2 versus T3) maintained its level of significance. There was a significant correlation (p less than 0.0001) between the pathological tumor size (maximum tumor diameter [(MTD) cm] and survival and DFI; once the MTD (less than versus greater than median value, 3.9 cm) was included in univariate and multivariate analysis, the tumor stage no longer appeared to maintain its level of significance. Treatment was generally well-tolerated and toxicity was acceptable. This study demonstrates the effectiveness of surgery and adjuvant chemotherapy in yielding impressive long-term survival for T1 and T2N0M0 SCLC and provides evidence that the MTD can be used as a powerful and independent prognostic factor for stratification of patients in further trials.

Adult↗
PubMed1991

Adjuvant chemotherapy for T1-2NOMO small cell lung cancer: single-agent or combination chemotherapy?

In an attempt to address the schedule of adjuvant chemotherapy in surgically resected T1 or T2NOMO small cell lung cancer, 12 patients were randomized to receive 6 courses of either single-agent (high-dose epirubicin) or combination (cyclophosphamide, epirubicin, and etoposide) chemotherapy, at 3-week intervals. No thoracic radiotherapy was administered while prophylactic cranial irradiation (30 Gy/10 fractions/2 weeks) was given. With a 25-month median followup, overall estimated 2-year and median survival were 83% and 26.5 months (range 16-34+), respectively. Ten patients are currently alive and disease free. No significant difference in 2-year survival was observed between the two adjuvant treatment modalities and median survival was 28 months (range 13-34+) for combination and 21 months (range 14-29+) for single-agent chemotherapy. Although at high doses, epirubicin resulted in a moderate clinical and histological cardiotoxicity and a remarkably reduced incidence of severe (WHO grades 3 and 4) treatment-related morbidity compared with the combination regimen. These preliminary results suggest that comparable survival and reduced toxicity might be expected with an active single-agent as adjuvant chemotherapy in T1 or T2NOMO small cell lung cancer.

Adult↗
PubMed1991

Increased invasive, chemotactic and locomotive abilities of c-Ha-ras-transformed human breast epithelial cells.

Transfection of the immortalized human breast epithelial cells MCF-10A with the mutated ras oncogene resulted in cell transformation (MCF-10A-neoT). Since the transformed state is usually associated with enhanced migratory activity, increased capability to invade basement membranes and to grow in a three-dimensional basement membrane gel (growth in matrigel), we compared these properties in MCF-10A-neoT cells with those of MCF-10A cells transfected with either the neomycin resistance gene alone (MCF-10A-neo cells) or with the normal ras proto-oncogene (MCF-10A-neoN cells). MCF-10A-neoT cells exhibited enhanced migratory activity, as assessed by chemotaxis and chemokinesis assays. and increased capability to invade the basement membrane. These cells also formed large colonies in matrigel. MCF-10A-neo and MCF-10A-neoN cells, on the other hand, showed only marginal migratory, invasive and semisolid medium growth properties. These results indicate that the mutated ras oncogene induces in human breast epithelial cells phenotypic characteristics of malignant transformation.

Adenocarcinoma↗
PubMed1990

Transfection of human breast epithelial cells with foreign DNA using different transfecting techniques.

The introduction of DNA into eukaryotic cells is a powerful technique for the study of gene regulation. This requires both a technique by which genes can efficiently be introduced into cells and a recipient cell representative of the tissue under study. We have utilized a spontaneously immortalized human breast epithelial cell line, CMF-10, which is phenotypically normal for introducing a neomycin-resistant gene contained in the plasmid Homer 6 (pHo6). Three different transfection methods were tested: calcium phosphate, with and without glycerol shock, using as control NIH/3T3 cells, and strontium phosphate and electroporation. In MCF-10 cells the efficiency of calcium phosphate mediated DNA transfection, which was measured as the number of colonies growing in neomycin-containing medium, was 21 and 140 fold higher than in MCF-10 cells transfected by electroporation and strontium phosphate methods, respectively. Glycerol shock enhanced three fold transfection efficiencies. NIH/3T3 transfected cells by calcium phosphate method showed a transfection efficiency similar to that of MCF-10 cells, however, glycerol shock did not improve the efficiency. These studies revealed that calcium phosphate combined with glycerol shock is the most efficient technique for transfection of genomic DNA into human breast epithelial cells.

Breast↗
PubMed1990

Fr-MLV infection induces erythroleukaemia instead of lymphoid leukaemia in mice given pituitary grafts.

Here we report that the slow-transforming helper component of Friend murine leukaemia virus (Fr-MLV), which produces lymphoid leukaemias in normal mice, induces erythroleukaemia in mice given syngeneic pituitary grafts (SPG). Newborn mice were infected with Fr-MLV and, at one month of age, were transplanted with two pituitary glands under the kidney capsule. Sham-operated infected mice and uninfected transplanted mice served as controls. SPG selectively reduced the mean survival times of infected mice. Histopathology showed that, while most infected non-transplanted mice developed lymphoid leukaemias, virtually all Fr-MLF-infected mice given SPG developed erythroleukaemias. Experiments in vitro showed that Fr-MLV infection markedly depressed concanavalin A induced DNA synthesis in cells from spleen, thymus and lymph nodes. Addition of prolactin or growth hormone further suppressed lectin-induced mitogenesis of lymphoid cells from infected mice, but failed to influence the response of uninfected controls. These experiments indicate that, in mice, pituitary hormones modulate the development and the histological features of Fr-MLV induced leukaemias, and suggest that endocrine-immunological interactions play a role in retrovirus induced tumorigenesis.

Animals↗