RESUMEN
BACKGROUND: Hormonally active environmental agents have recently been associated with the development of endometriosis. METHODS: We undertook a study to assess the relationship between endometriosis, an estrogen-dependent gynaecological disease, and 62 individual polychlorinated biphenyl (PCBs) congeners. We enrolled 84 eligible women aged 18-40 years undergoing laparoscopy for study, which included an interview and blood specimen (n=79; 94%). Thirty-two women had visually confirmed endometriosis at laparoscopy while 52 did not. Blood specimens were run in batches of 14 including four quality control samples for toxicological analysis. Each PCB congener was adjusted for recovery; batch-specific reagent blanks were subtracted. All PCB concentrations were log transformed and expressed in ng/g serum first as a sum and then as tertiles by purported estrogenic or anti-estrogenic activity of PCB congeners. RESULTS: Using unconditional logistic regression analysis, a significantly elevated odds ratio (OR) was observed for women in the third tertile of anti-estrogenic PCBs [OR 3.77; 95% confidence interval (CI) 1.12-12.68]. Risk remained elevated after controlling for gravidity, current cigarette smoking and serum lipids (OR 3.30; 95% CI 0.87-12.46). CONCLUSIONS: These data suggest that anti-estrogenic PCBs may be associated with the development of endometriosis.
Asunto(s)
Endometriosis/sangre , Endometriosis/etiología , Contaminantes Ambientales/sangre , Contaminantes Ambientales/toxicidad , Moduladores de los Receptores de Estrógeno/sangre , Moduladores de los Receptores de Estrógeno/toxicidad , Bifenilos Policlorados/sangre , Bifenilos Policlorados/toxicidad , Adolescente , Adulto , Estudios de Cohortes , Moduladores de los Receptores de Estrógeno/química , Femenino , Humanos , Oportunidad Relativa , Bifenilos Policlorados/química , Factores de RiesgoRESUMEN
Tumor angiogenesis, a major requirement for tumor outgrowth and metastasis formation, is regulated by pro- and anti-angiogenic factors. We have studied the expression of a panel of angiogenic factors, and of the angiogenesis inhibitor angiostatin, in a panel of human melanoma cell lines giving rise to xenografts with different vascular densities. Angiogenic-factor expression was analyzed in vitro (cell lines) and in vivo (xenografts), both at mRNA (RT-PCR and Northern blot) and at protein level (ELISA and Western blot). In vitro angiostatin generation was assessed by Western-blot analysis. Expression of bFGF and VEGF was clearly correlated with a high degree of vascularization, confirming the importance of these factors for tumor angiogenesis. In addition, there was exclusive or elevated in vitro expression of angiogenic factors IL-8, PDGF-AB, and, to a lesser extent, midkine in cell lines that formed highly vascularized tumors. A similar angiogenic-factor-expression pattern was found in the corresponding xenografts, with the exception of VEGF. In most cell lines, this factor had low expression in vitro which was strongly enhanced in vivo. Although all 8 melanoma cell lines were able to excise the angiostatin fragment from the plasminogen parent molecule in vitro, cell lines BLM and M14 showed the most potent angiostatin generation. In vitro angiostatin generation by cell lysates prepared from melanoma xenografts was comparable in all xenograft types. Thus, in our model system we found no correlation between angiostatin generation and vascular density. Our study has limited the number of pro-angiogenic factors that may be involved in melanoma angiogenesis, and provides evidence for the notion that regulation of tumor angiogenesis is dependent on multiple factors. Inhibition of angiogenesis for therapeutic purposes, therefore, should preferably not concentrate on a single factor.
Asunto(s)
Factores de Crecimiento Endotelial/análisis , Factor 2 de Crecimiento de Fibroblastos/análisis , Interleucina-8/análisis , Linfocinas/análisis , Melanoma/irrigación sanguínea , Neovascularización Patológica , Fragmentos de Péptidos/análisis , Plasminógeno/análisis , Factor de Crecimiento Derivado de Plaquetas/análisis , Angiostatinas , Animales , Factores de Crecimiento Endotelial/genética , Femenino , Factor 2 de Crecimiento de Fibroblastos/genética , Humanos , Interleucina-8/genética , Linfocinas/genética , Ratones , Ratones Endogámicos BALB C , Trasplante de Neoplasias , Fragmentos de Péptidos/biosíntesis , Fragmentos de Péptidos/genética , Plasminógeno/biosíntesis , Plasminógeno/genética , Factor de Crecimiento Derivado de Plaquetas/genética , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Trasplante Heterólogo , Factor A de Crecimiento Endotelial Vascular , Factores de Crecimiento Endotelial VascularRESUMEN
OBJECTIVE: To test the hypothesis that in women with polycystic ovary syndrome (PCOS), adrenal cytochrome P450c 17alpha activity is different after physiologic vs. pharmacologic ACTH stimulation and that ovarian activity promotes adrenal hyperactivity that is different after physiologic vs. pharmacologic ACTH stimulation. DESIGN: Prospective controlled pilot study. SETTING: Reproductive endocrinology unit of an academic medical center. PATIENT(S): Six women with PCOS who had adrenal hyperandrogenism were compared with four women with normal ovulation. INTERVENTION(S): Adrenal dynamic blood sampling was performed before and after 6 months of GnRH agonist administration. MAIN OUTCOME MEASURE(S): Comparison of physiologic and pharmacologic ACTH-stimulated levels of progesterone, 17-hydroxyprogesterone, and androgens before and after ovarian steroid modulation. RESULT(S): In women with PCOS, exaggerated responses of androstenedione and 11beta-hydroxyandrostenedione as well as elevated ratios of 17-hydroxyprogesterone to progesterone and of androstenedione to 17-hydroxyprogesterone after physiologic ACTH stimulation did not persist after GnRH-agonist administration. Three of the six women with PCOS had an increased response of androstenedione and a ratio of androstenedione to 17-hydroxyprogesterone that were >2 SD above the mean of those in the women with normal ovulation after pharmacologic ACTH stimulation; this finding persisted after GnRH-agonist administration. CONCLUSION(S): In women with PCOS, increases in adrenal androgen sensitivity after physiologic ACTH stimulation reflected in both arms of cytochrome P450c 17alpha activity may be influenced by ovarian activity. However, 17,20-lyase hyperactivity in a subset after pharmacologic ACTH stimulation may be an intrinsic adrenal disorder.
Asunto(s)
Hormona Adrenocorticotrópica/sangre , Cosintropina/uso terapéutico , Hormona Liberadora de Gonadotropina/agonistas , Síndrome del Ovario Poliquístico/tratamiento farmacológico , Síndrome del Ovario Poliquístico/enzimología , Esteroide 17-alfa-Hidroxilasa/metabolismo , Adulto , Femenino , Hormonas/sangre , Humanos , Hiperandrogenismo/complicaciones , Leuprolida/uso terapéutico , Ovario/fisiología , Proyectos Piloto , Síndrome del Ovario Poliquístico/sangre , Síndrome del Ovario Poliquístico/complicacionesRESUMEN
Through direct synthetic efforts we discovered a small molecule which is a 40 nanomolar inhibitor of the human FGF-1 receptor tyrosine kinase. 1-Tert-butyl-3-[6-(2,6-dichloro-phenyl)-2-(4-diethylamino-butylamino)-py rido[2,3-d]pyrimidin-7-yl]-urea (PD 161570) had about 5- and 100-fold greater selectivity toward the FGF-1 receptor (IC50 = 40 nM) compared with the PDGFbeta receptor (IC50 = 262 nM) or EGF receptor (IC50 = 3.7 microM) tyrosine kinases, respectively. In addition, PD 161570 suppressed constitutive phosphorylation of the FGF-1 receptor in both human ovarian carcinoma cells (A121(p)) and Sf9 insect cells overexpressing the human FGF-1 receptor and blocked the growth of A121(p) cells in culture. The results demonstrate a novel synthetic inhibitor with nanomolar potency and specificity towards the FGF-1 receptor tyrosine kinase.
Asunto(s)
Inhibidores Enzimáticos/farmacología , Pirimidinas/farmacología , Proteínas Tirosina Quinasas Receptoras/antagonistas & inhibidores , Receptores de Factores de Crecimiento de Fibroblastos/antagonistas & inhibidores , Urea/análogos & derivados , Animales , División Celular/fisiología , Línea Celular , Factores de Crecimiento de Fibroblastos/fisiología , Humanos , Fosforilación , Receptor Tipo 1 de Factor de Crecimiento de Fibroblastos , Receptores de Factores de Crecimiento de Fibroblastos/metabolismo , Proteínas Recombinantes/metabolismo , Spodoptera , Células Tumorales Cultivadas , Urea/farmacologíaRESUMEN
Basic fibroblast growth factor (bFGF) and other members of the FGF family share several biological properties that have the potential to mediate neoplastic cell growth. To test the hypothesis that bFGF may play a role in human ovarian cancer cell growth, three ovarian cancer cell lines, A90, A121(P), and A121(A), were investigated for their ability to respond to bFGF as a mitogen, to express endogenous bFGF protein or message for FGF proteins, and to exhibit FGF receptor or its message. Addition of bFGF to cultures of all three cell lines maintained in chemically defined media resulted in a statistically significant increase in cell number. Cell extracts from A90, A121(P), and A121(A) contained an immunoreactive protein that comigrated with hr-bFGF by Western blot analysis. Several bands of higher molecular weight were also noted. Immunohistochemical staining for bFGF demonstrated a cytoplasmic distribution of bFGF in the three cell lines. Both high- and low-affinity binding sites for human recombinant bFGF (hr-bFGF) were expressed by all three lines. High-affinity sites varied from 2700 sites per cell (Kd = 29 pM) to 13,500 sites per cell (Kd = 71 pM). All three cell lines were screened for mRNA expression for seven FGF proteins and four FGF receptors. In all three lines, mRNA for FGF2 (bFGF) was detected by PCR analysis, and in two lines, mRNA for FGF1 (aFGF) and FGF5 were also found. The FGFR1 receptor subtype (flg) was common to all of the cell lines. Finally, suramin inhibited proliferation of A90 and A121 (P and A) with IC50's of 60 and 210 micrograms/ml, respectively. This is consistent with the A90 cell line having higher levels of endogenous bFGF and flg and therefore being more responsive to suramin inhibition than the A121 cell line. The results indicate that these ovarian cancer cell lines can produce bFGF as well as other members of the FGF family of genes and have the ability to respond to bFGF.
Asunto(s)
Factor 2 de Crecimiento de Fibroblastos/análisis , Neoplasias Ováricas/química , Receptores de Factores de Crecimiento de Fibroblastos/análisis , Secuencia de Bases , Western Blotting , División Celular/efectos de los fármacos , ADN de Neoplasias/genética , Femenino , Factor 2 de Crecimiento de Fibroblastos/genética , Proteínas Filagrina , Regulación Neoplásica de la Expresión Génica , Humanos , Inmunohistoquímica , Datos de Secuencia Molecular , Neoplasias Ováricas/genética , Neoplasias Ováricas/patología , Reacción en Cadena de la Polimerasa , ARN Mensajero/análisis , ARN Mensajero/genética , Receptores de Factores de Crecimiento de Fibroblastos/genética , Suramina/farmacología , Células Tumorales CultivadasRESUMEN
In this study, we compared the cytotoxicity of cisplatin and carboplatin against a panel of human ovarian cancer cell lines using the MTT assay, a rapid colorimetric test that can be used to evaluate the number of residual viable tumor cells following chemotherapy. The established human ovarian cancer cell line OVCAR-3 and the recently isolated and characterized A721, A90, A286, A1, and A121A cell lines were evaluated for chemosensitivity. Each cell line was treated separately with cisplatin and carboplatin at concentrations ranging from 500 to 0.16 micrograms/ml. Various chemotherapeutic exposure periods (1, 4, 24, and 48 hr) were tested to determine maximal efficacy. All cell lines were more susceptible to cisplatin than carboplatin at all drug concentrations and all exposure periods tested (P = 0.005). The overall median 50% inhibitory concentration (ID50) for cisplatin was 107 micrograms/ml compared with 490 micrograms/ml for carboplatin P = 0.005). For both cisplatin and carboplatin a 24-hr exposure was significantly more cytotoxic than a 1-hr exposure (P = 0.003 and P = 0.006, respectively). These in vitro results suggest that cisplatin is significantly more cytotoxic than carboplatin against human ovarian cancer cell lines and that cisplatin should not be replaced by carboplatin in the treatment of advanced epithelial ovarian cancer until randomized trials using maximum dosing of the cisplatin-containing regimen are performed.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Neoplasias Ováricas/tratamiento farmacológico , Adulto , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Bleomicina/administración & dosificación , Bleomicina/efectos adversos , Cisplatino/administración & dosificación , Cisplatino/efectos adversos , Ciclofosfamida/administración & dosificación , Dactinomicina/administración & dosificación , Etopósido/administración & dosificación , Etopósido/efectos adversos , Femenino , Humanos , Neoplasias Ováricas/patología , Neoplasias Ováricas/cirugía , Vincristina/administración & dosificaciónRESUMEN
We report the cytogenetic findings in a primary endometrioid carcinoma of the ovary from a 29-year-old woman. Three clones with counts of 30, 29, and 27 chromosomes were observed. The predominant clone had 29 chromosomes and the following karyotype: 29, X, -X, -3, -3, +der(3)ins(3;?) (q21;?), -4, -5, -6, -8, -9, -11, -13, -14, -15, -16, -17, -18, -19, -21, -22, +mar. The clone with 30 chromosomes had an additional marker in the form of a minute chromosome, and the clone with 27 chromosomes was monosomic for chromosomes 12 and 20 also. Interestingly, there was no nullisomy of any of the chromosomes.
Asunto(s)
Adenocarcinoma/genética , Neoplasias Ováricas/genética , Adenocarcinoma/patología , Adulto , Femenino , Haploidia , Humanos , Cariotipificación , Neoplasias Ováricas/patologíaRESUMEN
Twenty-nine women with suspected pituitary adenomas were evaluated with nuclear magnetic resonance (NMR). Twenty-six had prolactin levels less than 100 ng/mL, and three had levels greater than 100. We tried to correlate the clinical findings with the prolactin levels and NMR findings. Pituitary adenomas were detected on NMR even with prolactin levels less than 100 ng/mL.
Asunto(s)
Adenoma/diagnóstico , Neoplasias Hipofisarias/diagnóstico , Adenoma/metabolismo , Adenoma/patología , Femenino , Hormona Folículo Estimulante/sangre , Humanos , Hormona Luteinizante/sangre , Imagen por Resonancia Magnética , Menstruación , Neoplasias Hipofisarias/metabolismo , Neoplasias Hipofisarias/patología , Prolactina/análisis , Estudios RetrospectivosRESUMEN
Six cell lines were established from four patients with advanced carcinoma of the ovary and from one patient with carcinoma of the endometrium. These lines were established from fresh tumor material maintained initially on culture dishes coated with an extracellular matrix (ECM) produced by bovine corneal endothelial cells. Two of the six lines continue to require ECM as a substrate for optimal growth while the remaining four lines will proliferate on ECM or plastic substrate. Four cell lines transplanted into athymic nude mice were tumorigenic and maintained histologic and karyotypic similarities between the patient's original tumor, the cell line, and the transplantable tumor. Furthermore, in vitro degradation of ECM was grossly apparent by those cell lines which formed nude mice xenografts. Tumor cells were characterized by cytology, transmission electron microscopy, karyology, substrate requirements, steroid binding protein analysis, and morphological appearance in culture.
Asunto(s)
Carcinoma/patología , Matriz Extracelular , Neoplasias Ováricas/patología , Células Tumorales Cultivadas , Neoplasias Uterinas/patología , Animales , Carcinoma/genética , Carcinoma/ultraestructura , Aberraciones Cromosómicas , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Trasplante de Neoplasias , Neoplasias Ováricas/genética , Neoplasias Ováricas/ultraestructura , Neoplasias Uterinas/genética , Neoplasias Uterinas/ultraestructuraRESUMEN
Although the concept of low malignant potential and/or borderline malignancy of some epithelial ovarian tumors was endorsed by the World Health Organization in 1973, uncertainty exists regarding the biologic behavior aspects of these lesions and this may account for the discrepancy in the 5-year survival figures reported for patients afflicted with these malignancies (76-95 per cent). We have reviewed the clinicopathologic aspects of 26 cases of borderline epithelial ovarian tumors and searched the literature. Based on our analysis, we have concluded that: 1) rupture of the cyst at surgery did not affect the patient's outcome but positive peritoneal fluid cytology did. 2) The term borderline should be replaced by ovarian intraepithelial neoplasia or preinvasive carcinoma and should solely be used in patients with stage I disease. 3) There is no justification for adjuvant therapy in adequately staged and surgically treated stage Ia and Ib disease. 4) Patients with stage II or more disease and those with positive peritoneal fluid cytology should be treated as aggressively as all other invasive, well-differentiated, epithelial ovarian tumors. 5) Our observation in cases of epithelial ovarian tumor cells grown on an extracellular matrix tends to indicate that parameters other than morphology may aid in assessing the invasive potential of these malignancies.
Asunto(s)
Cistadenocarcinoma/patología , Neoplasias Ováricas/patología , Adulto , Anciano , Anciano de 80 o más Años , Terapia Combinada , Cistadenocarcinoma/terapia , Diagnóstico Diferencial , Femenino , Humanos , Persona de Mediana Edad , Recurrencia Local de Neoplasia/patología , Estadificación de Neoplasias , Neoplasias Ováricas/terapiaRESUMEN
A cytogenetic analysis in a primary uterine leiomyosarcoma revealed a t(10;17) as the only chromosome change. This finding is discussed in relation to cytogenetic analyses on uterine leiomyomas and leiomyosarcomas reported previously.
Asunto(s)
Cromosomas Humanos Par 10 , Cromosomas Humanos Par 17 , Leiomiosarcoma/genética , Translocación Genética , Neoplasias Uterinas/genética , Adulto , Bandeo Cromosómico , Femenino , Marcadores Genéticos , Humanos , Cariotipificación , Leiomiosarcoma/patología , Neoplasias Uterinas/patologíaRESUMEN
The clinicopathologic aspects of seven cases of Fallopian tube adenocarcinoma are analyzed. Potential early spread of this malignancy to the retroperitoneal lymph nodes and right subdiaphragmatic area is documented. Multimodality treatment of tubal cancer to include surgery, radiation, and drug therapy (alkylating agents, progestins, with or without 5-fluorouracil and adriamycin) appears feasible and promising.
Asunto(s)
Adenocarcinoma/terapia , Neoplasias de las Trompas Uterinas/terapia , Adenocarcinoma/patología , Adulto , Anciano , Terapia Combinada , Neoplasias de las Trompas Uterinas/patología , Femenino , Humanos , Metástasis Linfática , Persona de Mediana Edad , Invasividad Neoplásica , PalpaciónRESUMEN
Forty patients with epithelial ovarian tumors underwent cyto-reductive surgery followed by a five drug (Adriamycin, D.D.P., 5FU, M.T.X. and C.T.X.) combination chemo and progestin therapy. They all had an initial complete clinical response and 58% were complete histologic responders. One patient developed reactivated disease six months after a negative second look laparotomy. Two of four fatal outcomes were due to development of mixed mesodermal tumors in patients who originally had serous cystadenocarcinomas. We conclude that this regimen is highly effective, independent of residual tumor size, histologic grade of tumor and prior therapy. Its attendant toxicity is acceptably low.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Neoplasias Ováricas/cirugía , Adulto , Anciano , Terapia Combinada , Femenino , Estudios de Seguimiento , Humanos , Persona de Mediana Edad , Estadificación de Neoplasias , Neoplasias Ováricas/tratamiento farmacológico , Neoplasias Ováricas/patología , ReoperaciónRESUMEN
Penetration of the extracellular matrix (ECM) by tumor cells, an event which occurs at various stages of the metastatic process, involves tumor cell glycosidase mediated hydrolysis of proteoglycans (PG). Recently, we observed that human ovarian carcinoma cell lines (HOCC) derived from primary tumors, peritoneal effusions, and distant metastases possess a varying ability to degrade radiolabeled PG of the ECM, while normal cells (human mesothelial cells or ovarian fibroblasts) fail to do so. To determine whether a quantitative relationship exists between glycosidase activity and degradation of ECM, both intracellular and extracellular glycosidase activities were measured for HOCC and normal cell lines. No relationship was found between intracellular glycosidase activities and the ability of cells to degrade ECM. However, a correlation was observed between extracellular or secretory glycosidase activities and HOCC mediated ECM degradation. In particular, a 5-8-fold increase, as compared to normal cells, was observed for HOCC extracellular beta-N-acetylglucosaminidase (EC 3.2.2.30) activity. The accumulation or secretion of this enzyme from HOCC into culture medium was found to be time dependent and not related to intracellular levels. Purified hexosaminidase derived from invasive HOCC was able to hydrolyze [3H]-glucosamine radiolabeled ECM (up to 30% radiolabel) and resulted in the cumulative release of free [3H]-N-acetylglucosamine. This enzyme mediated hydrolysis could be completely prevented with 2-acetamido-2-deoxy-1,5-D-gluconolactone, a competitive inhibitor (Ki 10(-6) M). Finally, HOCC mediated degradation of radiolabeled ECM was discerned to be dependent upon active hexosaminidase action, since tumor cell mediated degradation of ECM could be inhibited by up to 60% in the presence of this synthetic competitive inhibitor. In summary, these studies indicate a strong association between HOCC solubilization of glycoconjugates present in the ECM and extracellular levels of hexosaminidase.
Asunto(s)
Carcinoma/metabolismo , Matriz Extracelular/metabolismo , Glicósido Hidrolasas/metabolismo , Neoplasias Ováricas/metabolismo , Acetilglucosamina/metabolismo , Células Cultivadas , Femenino , Glucosamina/metabolismo , Humanos , Concentración de Iones de Hidrógeno , Cinética , Inhibidores de Proteasas/farmacología , beta-N-Acetilhexosaminidasas/farmacologíaRESUMEN
Better in vitro models are needed to elucidate the mechanisms underlying tissue destruction by human tumor cells. To address this matter recently isolated and characterized human ovarian carcinoma cell lines derived from either primary tumors, ascitic effusions or metastatic growths were plated in direct contact with extracellular matrix (ECM) previously deposited on culture dishes by bovine corneal endothelial cells. Light and electron microscopy of four of the five ovarian tumor cell lines demonstrated morphologic digestion with penetration of ECM by tumor cell microvilli, along with associated rarefaction. The ability of these same ovarian tumor cell lines to solubilize specific carbohydrate and protein moieties present in intact ECM was assessed with the use of metabolically prelabeled ECM employing tritiated fucose, galactose, glucosamine and proline. Results from these studies corroborated morphologic observations in which four of the five tumor cell lines tested extensively solubilized radiolabeled ECM. The kinetics of radiolabel release from ECM illustrated that three of the four invasive tumors released [3H]fucose, [3H]glucosamine and [3H]proline at high rates. Normal human ovarian fibroblasts and mesothelial cells were observed to be unable to digest ECM and this was consistent with their inability to release radiolabeled material from prelabeled ECM. The results from these studies suggest that some ovarian carcinomas have the ability to degrade basement membrane components. Knowledge regarding the mechanisms responsible for tissue degradation may eventually lead to the development of new chemotherapeutic modalities designed to restrict tumor cell invasion, growth and metastasis.
Asunto(s)
Matriz Extracelular/ultraestructura , Neoplasias Ováricas/ultraestructura , Células Cultivadas , Femenino , Humanos , Técnicas In Vitro , Cinética , Microscopía Electrónica de Rastreo , Microscopía de Contraste de Fase , SolubilidadRESUMEN
Four cases of advanced stage (II or III) and one case of early stage (IC) borderline malignant serous cystadenocarcinomas of the ovary were maintained on culture dishes coated with an extracellular matrix (ECM) produced by bovine corneal endothelial cells. Cells harvested for chromosomal analysis after 2-3 days showed diploid or near-diploid modalities in all cases. Banded chromosome studies in two cases revealed nonrandom clonal abnormalities with trisomy 2, 7, and 12 in seven of 13 metaphases. No structural abnormalities were noted. These cytogenetic findings differ from those found in malignant serous tumors of the ovary. In addition, borderline tumor cells digested the ECM in all cases and formed a cribiform pattern within a few days of primary culture. This study suggests clonal progression from early to advanced stages of borderline malignant serous tumors; readily distinguishable from overtly malignant serous tumors of the ovary. Ability of tumor cells derived from both primary tumors and metastatic implants to digest the ECM implies the possibility that borderline serous tumors have invasive potential.
Asunto(s)
Neoplasias Ováricas/patología , Adulto , Anciano , Células Cultivadas , Bandeo Cromosómico , Femenino , Humanos , Cariotipificación , Persona de Mediana Edad , Invasividad Neoplásica , Metástasis de la Neoplasia , Estadificación de Neoplasias , Neoplasias Ováricas/genéticaRESUMEN
Human mesothelial cells (HMC) cover a variety of serosal surfaces and have been shown to rest upon an underlying subcellular basement membrane in vivo. Bovine corneal endothelial cells produce an extracellular matrix (ECM) in vitro that mimics HMC subcellular basement membrane and was found to modulate HMC adhesion, morphology and proliferation in vitro. Our results indicated that within minutes after plating, a high percentage (greater than 80%) of HMC firmly attached to ECM. Active cellular migration and subsequent proliferation were observed leading to the formation of a well-organized closely apposed cell monolayer. However, when cells were plated on plastic, the rate of cell attachment was much lower and the proliferative rate of HMC grown on plastic also was strikingly lower (exponential doubling time 4.3 days) than that of cells grown on ECM (exponential doubling time 2.4 days). Cells upon reaching confluency on plastic were markedly enlarged as compared to confluent cells grown on ECM. These observations corroborated differences in final cell density where it was noted that HMC cultured on ECM demonstrated a 10-fold greater final cell density as compared to cells grown on plastic. Results from these studies illustrate the fact that phenotypic expression as well as proliferative responsiveness of HMC can be modulated by adhesive interactions with preformed ECM.
Asunto(s)
Células Epiteliales , Adhesión Celular , División Celular , Células Cultivadas , Epitelio/ultraestructura , Matriz Extracelular , Humanos , Microscopía Electrónica de RastreoRESUMEN
Culture dishes coated with an extracellular matrix (ECM) produced by bovine corneal endothelial cells were utilized to investigate human gynecologic carcinomas of epithelial origin. A high culture success rate was achieved for both solid tumor (83%) and ascitic fluid (75%) derived from specimens from 59 patients. Because of the high percentage of tumor cells attaching to the ECM and actively proliferating, a variety of in vitro studies (karyotypic analysis, morphologic appearance on ECM, and ability to digest the ECM) could be done. A preliminary in vitro profile of the various tumor cell types could be made on the basis of these studies. In addition, five long-term cultures were established utilizing the ECM model system.
Asunto(s)
Carcinoma/ultraestructura , Matriz Extracelular/ultraestructura , Neoplasias de los Genitales Femeninos/ultraestructura , Adenocarcinoma/genética , Adenocarcinoma/ultraestructura , Carcinoma/genética , Células Cultivadas , Femenino , Neoplasias de los Genitales Femeninos/genética , Humanos , Cariotipificación , Modelos Biológicos , Neoplasias Ováricas/genética , Neoplasias Ováricas/ultraestructura , Neoplasias Uterinas/genética , Neoplasias Uterinas/ultraestructuraRESUMEN
The major obstacle to successful cytogenetic analysis of human solid tumors is the acquisition of sufficient numbers of good quality metaphases for detailed cytogenetic analysis. At present, no single methodologic approach has been proven to provide successful chromosomal analysis of all human solid tumors. The technical aspects of cell culture, chromosome harvesting, and chromosome banding were the focus of considerable discussion during the First Workshop on Chromosomes in Solid Tumors. This report provides summaries of several technical protocols, emanating from several different laboratories, which have contributed to successful chromosome analysis of a variety of human solid tumors.
Asunto(s)
Células Cultivadas , Bandeo Cromosómico/métodos , Neoplasias/genética , Separación Celular , Humanos , Neoplasias/patologíaRESUMEN
Human ovarian tumors metastasize by direct extension into the peritoneal cavity leading to tumor cell implantation onto peritoneal surfaces. Successful formation of peritoneal implants is dependent on the ability of ascitic tumor cells to infiltrate the mesothelium, and become firmly adherent to the underlying extracellular matrix (ECM). In order to investigate this process in more detail, an in vitro model system was developed employing human mesothelial cells grown on ECM-coated culture dishes. The ability of human ovarian carcinoma cells derived from ascitic fluid to attach to the mesothelial cell monolayer grown on ECM, ECM alone or plastic was quantitated with the use of 51Cr radio-labelled tumor cells. Tumor cells exhibited a more rapid and firmer attachment to ECM than to the mesothelial cells or to plastic. Using agitation to stimulate peritoneal fluid dynamics and shear forces in vivo, tumor cell arrest was found to be limited to the ECM, but it occurred at a slower rate than it did without agitation. Tumor cell attachment was also restricted to areas of exposed ECM in wounded mesothelium as assessed by phase-contrast microscopy. Morphologic alterations of the mesothelium induced by tumor cells were observed with the use of scanning electron microscopy (SEM) and immunohistochemical staining which included disruption of intercellular junctions leading to retraction of mesothelial cells, exposure of underlying ECM, subsequent attachment and proliferation on ECM. This model system would appear to be useful for elucidating mechanisms of ovarian tumor cell adhesion and proliferation, and for assessing various therapeutic modalities for their ability to block tumor cell implantation, invasion and growth on peritoneal surfaces.