Progression in teratocarcinomas.
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Aneuploidy↗
Biomedical subjects
G B Pierce
Publications and source records attributed to G B Pierce.
Hydrogen peroxide as a mediator of programmed cell death in the blastocyst.
Previous work identified in blastocele fluid a soluble activity which killed embryonal carcinoma cells with trophectodermal potential but not those with embryonic potential [35]. From use of a malignant caricature of the late blastocyst, this toxic activity was postulated to be H2O2 [8]. The purpose of this paper was to determine if blastocele fluid also contained amounts of H2O2 capable of mediating the preferential killing of malignant pretrophectodermal cells (ECa 247). We not only observed that blastocele fluid is not toxic for these cells in the presence of catalase, but that malignant cells with embryonic potential (P19) that normally survive exposure to blastocele fluid become sensitive to it if their intracellular glutathione levels are lowered. Thus, it is concluded that the blastocyst contains amounts of H2O2 toxic to malignant pretrophectodermal cells and that glutathione-dependent mechanisms protect malignant inner cell mass cells with embryonic potential. Apparently, H2O2 production and glutathione-dependent protection mechanisms are developmentally regulated in the inner cell mass. These results are discussed with regards to apoptosis and the regulation of tissue mass.
Animals↗
Amine oxidases, programmed cell death, and tissue renewal.
Embryonal carcinoma cells, with embryonic (ECaE) or trophectodermal (ECaT) potential, have been used in a colony assay to determine regulatory mechanisms in the blastocyst. The mechanism that regulates ECaE and results in chimera formation is dependent upon a soluble factor in blastocoele fluid and contact with trophectoderm. Two mechanisms contribute to the regulation of ECaT: one involves a factor in blastocoele fluid and the other contact with either trophectoderm or inner cell mass which results in differentiation of the cells into trophectoderm, and the other involves the killing of at least 40% of the cells by blastocoele fluid alone. This cytotoxic activity probably causes the programmed cell death that occurs in the inner cell mass during blastulation as it loses the potential to differentiate into trophectoderm. A toxic activity similar to that of normal blastocysts has been obtained from embryoid bodies. This activity is caused by amine oxidase-dependent catabolism of polyamines, and it is postulated that programmed cell death in the embryo and chalone activity in the adult may have similar mechanisms.
Animals↗
Serum amine oxidase activity contributes to crisis in mouse embryo cell lines.
This paper reports the results of experiments to test the hypothesis that crisis of spontaneous transformation is caused by the hydrogen peroxide and/or aldehydes generated from endogenous polyamines by serum amine oxidase [amine: oxygen oxidoreductase (deaminating), EC 1.4.3.6]. After 4-5 weeks of culture, crisis occurred in 16 of 29 cell lines derived from limb buds of embryos from SJL/J, C3H, and CD-1 mice. In contrast, after the same time in culture but in medium supplemented with aminoguanidine, which inhibits serum amine oxidase, crisis occurred in only 1 of 41 cell lines. Protection against crisis was maximal in cell lines of SJL/J embryos, in which the incidence of crisis fell from 7 of 9 in untreated controls of 0 to 12 in the presence of 2 mM aminoguanidine. 2-Mercaptoethanol at 150-300 microM, which protects cells from serum amine oxidase-dependent polyamine toxicity, also protected the cell lines against crisis. These protected cell lines retained proliferative potential, diploid DNA content, and the mixture of cell types found in the primary cultures. These results indicate that cytotoxic catabolites generated by serum amine oxidase caused at least a large portion, but perhaps not all, of the cellular damage that leads to crisis in mouse embryo cell lines.
Aldehydes↗
Neoplastic embryoid bodies of embryonal carcinoma C44 as a source of blastocele-like fluid.
There is a cytotoxic activity in blastocele fluid that kills embryonal carcinoma cells with trophectodermal potential but spares those with embryonic potential. This activity is present when programmed cell death occurs in the inner cell mass (ICM), and the ICM loses its trophectodermal potential. Because of the paucity of blastocele fluid, cystic embryoid bodies of embryonal carcinoma C44 were examined ultrastructurally and in tissue culture to determine if they corresponded to late blastocysts and if their fluid corresponded to blastocele fluid. No trophectoderm was demonstrated in the embryoid bodies, but embryonal carcinoma and endoderm were present, leading to the conclusion that the embryonal carcinoma corresponded to late ICM that had expressed endodermal potential. As a result the cyst fluid might have contained the toxic activity of blastocele fluid. The cyst fluid of C44 embryoid bodies did contain a soluble, low-molecular-weight, cytotoxic activity that preferentially killed embryonal carcinoma cells with trophectodermal potential while sparing those with embryonic potential. Enough of this fluid was available to determine the chemical nature of this toxic activity.
Animals↗
Evidence linking programmed cell death in the blastocyst to polyamine oxidation.
Programmed cell death occurs in the inner cell mass during blastulation concomitant with the loss of its trophectodermal potential, and blastocele fluid kills malignant inner cell mass cells with trophectodermal potential (ECa 247) but spares those with embryonic potential (P19). A previous study had shown that blastocele-like fluid from embryoid bodies of the teratocarcinoma C44 contains a low-molecular-weight cytotoxin that exhibits the same target-cell selectivity as normal blastocele fluid. The current paper shows that the preferential killing of cells with trophectodermal potential is caused by hydrogen peroxide generated during the oxidation of polyamines in the cyst fluid by amine oxidases. The greater resistance of cells with embryonic potential to hydrogen peroxide is due to glutathione-dependent mechanisms. These data lead to the conclusion that an amine oxidase in the blastocyst oxidizes polyamines in blastocele fluid, generating hydrogen peroxide which causes programmed cell death of normal and malignant cells with trophectodermal potential.
Animals↗
Embryonic mechanisms for abrogating the malignancy of cancer cells.
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Animals↗
Polyamine oxidation, programmed cell death, and regulation of melanoma in the murine embryonic limb.
The murine embryonic limb at day 14 of gestation suppresses tumor formation by melanoma cells. Conditioned media of embryonic limbs have been found cytotoxic for B16 melanoma cells. The cytotoxicity is due to the catabolism of polyamines in the limb bud extracts by an amine oxidase in the serum supplement of the culture medium. However, a polyamine oxidase activity, similar to that in adult rat liver, is also detectable in homogenates of embryonic limbs. Thus, the embryonic limb contains the necessary components to produce polyamine-derived cytotoxic metabolites, which are present at the time programmed cell death occurs. This leads to the hypothesis that injected melanoma cells are killed incidentally by the mechanism that mediates programmed cell death.
Amine Oxidase (Copper-Containing)↗
Mechanism of programmed cell death in the blastocyst.
The malignant growth potential of embryonal carcinoma cells may be controlled by environmental factors. For example, embryonal carcinoma cells placed into normal blastocysts may not exhibit the continued growth expected of malignant cells but rather may lose all aspects of the malignant phenotype and become apparently normal embryonic cells. Loss of the malignant phenotype of embryonal carcinoma cells occurs early in these injected blastocysts and has been used as the basis of assays to study the mechanisms of regulation of embryonal carcinoma by the blastocyst. In this regard, P19, an embryonal carcinoma that makes midgestation chimeras, was regulated by blastocele fluid plus contact with trophectoderm but not by blastocele fluid plus contact with inner cell mass (ICM). In contrast, ECa 247, which makes trophectoderm, was regulated by exposure to blastocele fluid plus contact with trophectoderm or ICM. During the course of these experiments, dead embryonal carcinoma and ICM cells were observed, and blastocele fluid was then shown to kill ECa 247 and normal ICM cells of early blastocysts with trophectodermal potential. P19 cells and ICM cells with potential to make the embryo were not killed by blastocele fluid. Programmed cell death occurs in the ICM of the blastocyst during the transition from early (when ICM has the potential to make trophectoderm) to late (when the ICM lacks the potential to make trophectoderm). It is postulated that this programmed cell death is designed to eliminate redundant ICM cells with trophectodermal potential, and its mechanism of action is mediated by epigenetic factors in blastocele fluid.
Animals↗
Programmed cell death in the blastocyst.
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Animals↗
Tumors as caricatures of the process of tissue renewal: prospects for therapy by directing differentiation.
A concept of neoplasms, based upon developmental and oncological principles, states that carcinomas are caricatures of tissue renewal, in that they are composed of a mixture of malignant stem cells, which have a marked capacity for proliferation and a limited capacity for differentiation under normal homeostatic conditions, and of the differentiated, possibly benign, progeny of these malignant cells. The concept brings order to the facts about carcinoma, has predictive value for embryogenesis, and indicates possibilities for differentiation therapy. One such possibility assumes on the basis of experimentation in vitro that malignant stem cells can be induced to differentiate into postmitotic cells by application of chemicals. Another suggests study of naturally occurring substances which regulate cell proliferation and differentiation in adult tissues. The other possibility, based upon experiments in vivo and in vitro, indicates that embryonic fields are capable of converting their closely corresponding malignant lineages into apparently normal lineages responsive to homeostatic control. Induced differentiation of embryonal carcinoma has been achieved in vivo with improvement in longevity of the host and in some cases with apparent cure. However, ultimate success of treatment based upon turning malignant cells into benign cells will depend upon the nature of the benign cells. Will they remain benign?
Azacitidine↗
Tumorigenicity of embryonal carcinoma as an assay to study control of malignancy by the murine blastocyst.
A bioassay, based on tumorigenicity, has been developed to determine the mechanism whereby the blastocyst of the mouse controls malignant expression of embryonal carcinoma. The assay is based upon the incidence of tumors obtained when known numbers of cells of the 402AX strain of embryonal carcinoma are injected into strain 129 mice, compared to the incidence obtained when the same number of embryonal carcinoma cells are incorporated into Swiss-Webster blastocysts that are then injected in strain 129 animals. The results indicate that the blastocyst can regulate one embryonal carcinoma cell consistently; it may have a slight effect on three, but it cannot regulate four or five of them. The position of the embryonal carcinoma cell in the blastocyst is important. Regulation occurs if the embryonal carcinoma cell is placed in the blastocoele cavity, but enhancement of tumorigenicity is obtained if it is placed between the zona pellucida and the trophectoderm. By contrast, the blastocyst is unable to regulate a single B-16 melanoma cell placed in the blastocoele cavity, indicating a degree of specificity for the regulatory process.
Animals↗
Yolk sac carcinoma (endodermal sinus tumor): ultrastructure and histogenesis of gonadal and extragonadal tumors in comparison with normal human yolk sac.
Human yolk sac carcinomas have been studied only twice with the electron microscope, and have never been compared at this level with normal human yolk sac. In the present study, the ultrastructural features of three primary ovarian yolk sac carcinomas, omental metastases from one of these, and a primary retroperitoneal yolk sac carcinoma in a male are reported, as are the ultrastructural findings in human yolk sac from normal 7- and 12-week gestations. The most prominent feature of the tumors is the presence of voluminous basement membrane material (the nature of which is confirmed by indirect enzyme-labeled antibody technique in one case) in both intra-and extracellular location, corresponding to the PAS-positive hyaline globules seen in these tumors by light microscopy. The tumor cells are also demonstrated to produce this material in tussue culture. Although basement membrane has not been described previously in normal human yolk sac at 8 and 10 weeks' gestation, it was present in the 7-week specimen which we studied, suggesting that its production may be a feature of only very young sac. Other ultrastructural findings are also similar in human yolk sac carcinoma, normal human yolk sac, and rodent yolk sac and yolk sac carcinomas. Thus, these studies confirm the suggested germ cell-derived yolk sac origin of the human tumor.
Adult↗
Relationship between differentiation and carcinogenesis.
Carcinomas are caricatures of the normal process of tissue renerwal. Malignant stem cells proliferate, and some of their progeny differentiati and form benign functional cells. In teratocarcinoma, it has been demonstrated that the stem cells are the target in carcinogenesis and become malignant stem cells. The normal and malignant stem cells are equally differentiated. Normal stem cells of breast and colon are no more differentiated than their counterparts. If they are the target in carcinogenesis, then the concept of dedifferentiation is bypassed as an explanation for the undifferentiated appearance of tumors. While the focus of this meeting has been on mutation as an explanation for carcinogenesis, in this paper emphasis is placed on electrophilic carcinogens acting on cytoplasmic molecules that control gene expression. A type of gene control in addition to the operon is postulated.
Animals↗
Ultrastructural comparison of differentiation of stem cells of murine adenocarcinomas of colon and breast with their normal counterparts.
Two rats with chemically induced transplantable adenocarcinomas of the colon were given pulses of [3H]thymidine, and autoradiography with electron microscopes was used to compare the degrees of differentiation of the stem cells of the tumor and colon. The best differentiated portions of the tumor had acini composed of vacuolated, mucous, and argentaffin cells in various stages of differentiation. Vacuolated and mucous cells incorporated [3H]thymidine and corresponded in degree of differentiation to that of their labeled normal counterparts in the normal colon. An exceedingly undifferentiated labeled cell, hitherto undescribed, was identified in the tumor and crypts of the colon; this may be an undifferentiated colon stem cell that differentiates into vacuolated and mucous stem cells and/or into argentaffin cells. Normal stem cells of the breast and malignant stem cells of spontaneous adenocarcinomas of the breast of C3H mice had comparable degrees of differentiation. Since normal stem cells in these tissues were as undifferentiated as the least differentiated stem cells of the tumors, there is now no need to postulate dedifferentiation as a mechanism to explain the undifferentiated appearance of tumors.
Adenocarcinoma↗
Latent carcinoma and carcinoma in situ.
Lesions exist in the cervix that are diagnosed as carcinoma in situ; some may progress to invasive carcinoma and some may regress. Many are probably overdiagnosed, may represent effects of promoting agents rather than intiating agents, and may entail risk for the patient in that the lesion may be an unusualy good target in carcinogenesis. Latent carcinomas are small foci of initated cells unable to express their malignant phenotype because of environmental controls. Latent cells are produced in experimental carcinogenesis and occur in spontaneous tumors as G-O stem cells.
Adenocarcinoma↗
Removal of basement membrane in the involuting breast.
Morphologic and immunohistochemical studies by light and electron microscopy indicated that basement membrane was removed during the process of involution of the murine breast. Removal of the basement membrane started 2 days postweaning, was maximal at 4 days, and correlated with degeneration of epithelial cells. There was no evidence of phagocytosis of basement membrane, so the removal of this antigen was attributed to enzymatic hydrolysis. To determine the activity of breast homogenate on the specific basement membrane antigen, insoluble basement membrane embedded in agarose gels was incubated with breast liver and kidney homogenates. When basement membrane antigen was demonstrated by the specific antibody, it was found that breast homogenate solubilized basement membrane but liver and kidney failed to solubilize basement membrane. To quantify the reaction and determine some of the characteristics of the responsible enzyme(s), insoluble basement membrane was labeled with 125I and the release of radioactivity into the supernatant following incubation with extracts of involuting breast indicated hydrolysis of basement membrane. Extracts of breast homogenate extensively hydrolyzed labeled basement membrane if naturally occurring inhibitors were removed by previous washing, whereas liver or kidney extracts prepared in a similar manner were devoid of activity. The hydrolysis of basement membrane was time and concentration dependent and had a pH optimum. The reaction was blocked by prior heating of the extract at 100 degrees C. for 30 minutes, removal of divalent cations, and presence of diisopropylfluorophosphate (a specific serine esterase inhibitor); prolonged dialysis failed to remove the hydrolytic activity. It is concluded that an enzyme system present in the involuting breast is capable of basement membrane hydrolysis
Animals↗