High-metastatic clones selected in vitro from a recent spontaneous BALB/c mammary adenocarcinoma cell line.

The metastatic TS/A line has been recently derived from a spontaneous BALB/c mammary tumor. When TS/A cells were cultured in 0.33 per cent agar, two morphologically distinct types of colonies were observed from which two sets of clones were obtained.

Impaired H-2 expression in B16 melanoma variants.

We studied the expression of H-2b alloantigens in three different B16 melanoma lines cultures in vitro. Cell lines were B16-F1 and two cell cultures (named B16-A and B16-B) newly derived from two different in vivo sublines of B16 melanoma. The assays used were in vivo tumour growth in allogeneic (BALB/c and B10.

TS/A: a new metastasizing cell line from a BALB/c spontaneous mammary adenocarcinoma.

A metastasizing mouse cell line (TS/A), originated from a mammary adenocarcinoma which arose spontaneously in a BALB/c female retired breeder, has been established in vitro. It displayed a remarkable morphologic heterogeneity, which is evident in plastic adherent cultures (cell types ranging from epithelial-like to fibroblast-like) as well as in semi-solid agar cultures. The TS/A line exhibited the presence of specific cytoplasmic estradiol receptor, with a binding activity of 16 fmoles/mg cytosol protein.

Inverse relationship between anti-SV40 TASA and anti-H-2 cytotoxic responses.

The in vitro cytotoxic response against H-2d and H-2b SV40-transformed fibroblasts was studied in a 40-h 3H-proline assay. A very low response against SV40 TASA is associated with the H-2d antigens on target cells: however, SV40-transformed H-2d cells are as immunogenic as SV40-transformed H-2b cells and prime against H-2b target cells. The data concerning in vitro amplification of the anti-SV40 TASA response and the involvement of cyclophosphamide-sensitive suppressor populations confirm the comparable immunogenicity of SV40-transformed H-2d AND H-2b cells and cannot account for the haplotype-related behavior observed with SV40-transformed target cells.

Glucocorticoid receptor and in vitro sensitivity to steroid hormones in human lymphoproliferative diseases and myeloid leukemia.

The glucocorticoid receptor (GR) quantitation by a whole-cell assay and/or cytosol technique and the in vitro sensitivity to steroids have been assessed in peripheral blood cells from normal donors and patients with chronic lymphatic leukemia (CLL), acute lymphoblastic leukemia (ALL), lymphosarcoma cell leukemia (LSCL), acute nonlymphatic leukemia (ANLL), and chronic myeloid leukemia (CML). Within the lymphoproliferative diseases, ALL cells exhibited the highest GR concentration (regardless of the method used) and the highest in vitro inhibition of spontaneous [3H]thymidine ([3H]TdR) uptake by glucocorticoids. A significant relationship between GR concentration (whole-cell assay) and in vitro sensitivity to dexamethasone was also found.

The occurrence of multiple steroid hormone receptors in disease-free and neoplastic human ovary.

The cytoplasmic receptors for 17 beta-estradiol (ER), 5 alpha-dihydrotestosterone (AR), progesterone (PR), and cortisol (GR) have been quantified in 36 specimens from the human ovary (13 disease-free, 5 benign, and 18 malignant) by a dextran-coated charcoal (DCC) technique. The occurrence of receptor-positive biopsies were: ER 46%, AR 85%, PR 54%, GR 92%, in normal tissue; ER 40%, AR 100%, PR 20%, GR 50%, in benign tumors; and ER 67%, AR 72%, PR 50%, GR 88%, in malignant lesions. Furthermore, the simultaneous occurrence of ER and PR in malignant tumors was 50% yet all four receptors were found to be present only in 44% of the cases.

Correlation between clinical response to antiandrogenic therapy and occurrence of receptors in human prostatic cancer.

Occurrence of steroid hormone receptor has been evaluated in 30 prostatic cancer-bearing patients: 5 alpha-dihydrotestosterone receptor (DHTR) occurrence was correlated with both tumor grade of differentiation and clinical response to hormone therapy.

Multiple steroid hormone receptors in normal and abnormal human endometrium.

The cytoplasmic concentrations of ER, AR, PR, and GR were determined in 124 specimens of normal and abnormal endometrium and other uterine human tissues by the DCC technique. In the endometrial carcinoma group, we observed that pretreatment with MAP leads to low cellularity, higher amount of AR, lower amounts of detectable ER, GR, and PR: the last receptor was almost always absent. A positive correlation between ER presence and tumor grade of differentiation was found in endometrial tumors from hormone-untreated patients.

17 beta-estradiol, 5 alpha-dihydrotestosterone, progesterone and cortisol receptors in normal and neoplastic human endometrium.

Using the value of 0.24 fmoles/microgram DNA as breaking point between high and low binding capacities, we quantified receptors for 17 beta-estradiol (ER), 5 alpha-dihydrotestosterone (DHTR), progesterone (PR) and cortisol (CR) in normal and neoplastic human uterine tissues. Concerning receptors occurrence, significant relationships were observed between ER and PR, ER and DHTR, and DHTR and PR.

[Measurement of cytoplasmic receptors specific for steroid hormones. IV) Glucocorticoid receptor].

Binding of 3H-cortisol (3H-C) and 3H-dexamethasone (3H-DX), a fluorinated glucocorticoid which does not bind to Corticosteroid Binding Globulin (CBG), was investigated in two different target tissues, human breast cancer and human leukemic lymphocytes, using a modified DCC assay method. Maximal binding to cytosol occurred, when increasing concentrations of 3H-C or 3H-DX were incubated in the presence or in the absence of an excess of cold cortisol or dexamethasone, at 0 degrees C for 1 hr or 4 hr, respectively. A 50 sec exposure to DCC was sufficient to separate bound from free labelled steroid.

[Determination of cytoplasmic receptors for specific steroid hormones. III) Progestin receptor].

A method for determining cytoplasmic progesterone receptor was standardized in normal human endometrium comparing two different tracers, 3H-progesterone (3H-P) and 3H-medroxyprogesterone acetate (3H-MAP), a synthetic progestin which does not bind to Corticosteroid Binding Globulin (CBG). Receptor assays were performed as previously reported for 17beta-estradiol receptor, with slight modifications: incubation lasted 1 hr at 0 degree C, followed by 5 min DCC exposure under the same conditions. When 3H-P was employed as tracer, blanks performed with cold MAP gave similar results as using cortisol in incubation tubes and progesterone and cortisol in blanks.

[Determination of cytoplasmic receptors for specific steroid hormones. II) Androgen receptor].

Cytoplasmic receptors for 5 alpha-dihydrotestosterone (3H-DHT) were determined in normal and hypertrophic human prostate using the slightly modified DCC method we previously standardized for 17beta-estradiol-receptor. Incubations were always performed at 0 degree C for 1 hr. Discrimination between 3H-DHT binding to cytoplasmic receptor and to Sex Hormone Binding Globulin (SHBG) was achieved on the basis of binding affinity, thermolability and pattern of specificity by various steroid hormones.

In vivo reaction of dimethylnitrosamine with nucleic acids.

7-Methylguanine is the main compound resulting from in vivo interaction between dimethylnitrosamine (DMNA) and nucleic acids, detected after strong acid hydrolysis. However, enzymic and alkaline hydrolysis of nucleic acids leads to quantitative liberation of methylamine. Methylamine isolated from liver nucleic acids of 15N-DMNA-treated rats has molecular weight of 31, thus demonstrating that DMNA-nitrogen is not involved in the binding.