Characterization of TIS7, a gene induced in Swiss 3T3 cells by the tumor promoter tetradecanoyl phorbol acetate.

We report here the nucleic acid sequence and deduced amino acid sequence of a cDNA for TIS7, a gene induced by mitogens in 3T3 cells and by nerve growth factor in PC12 pheochromocytoma cells. A fragment of the TIS7 gene has previously been cloned from murine fibroblast cells infected with Newcastle Disease Virus. The cDNA sequence suggests some similarities with murine interferons.

Induction of c-fos and TIS genes in cultured rat astrocytes by neurotransmitters.

The interaction of neurotransmitters with their specific receptors initiates a cascade of intracellular biochemical events which lead to induction of specific genes. Included in this cascade is the rapid and transient induction of a family of primary early response genes we term TIS genes (Lim et al.: Oncogene 1: 263-270, 1987).

The tumor promoters 12-O-tetradecanoylphorbol-13-acetate and okadaic acid differ in toxicity, mitogenic activity and induction of gene expression.

Okadaic acid (OA) and 12-O-tetradecanoylphorbol-13-acetate (TPA) are both potent tumor promoters in a mouse skin carcinogenesis experiment. OA was much more toxic than TPA for murine embryo cell lines such as Swiss 3T3 cells or C3H10T1/2 cells. TPA is a potent mitogen for 3T3 cells; in contrast OA was unable to stimulate DNA synthesis in these cells.

Granulocyte-macrophage colony-stimulating factor and tetradecanoyl phorbol acetate induce a distinct, restricted subset of primary-response TIS genes in both proliferating and terminally differentiated myeloid cells.

Induction of early-response genes (tetradecanoyl phorbol acetate [TPA]-induced sequences, or TIS genes; R.W. Lim, B.

TIS gene expression in cultured rat astrocytes: multiple pathways of induction by mitogens.

Accumulation of TIS1 and TIS11 (Lim et al.: Oncogene 1:263-270, 1987) mRNAs in secondary cultures of rat neocortical astrocytes was much greater in response to tetradecanoyl phorbol acetate (TPA) than in response to either epidermal growth factor (EGF) or fibroblast growth factor (FGF). In contrast, EGF, FGF, and TPA were equally effective in inducing accumulation of TIS8 and TIS28/c-fos mRNAs.

TIS gene expression in cultured rat astrocytes: induction by mitogens and stellation agents.

The expression of a number of TIS genes (Lim et al.: Oncogene 1:263-270, 1987) was examined in secondary cultures of rat neocortical astrocytes treated with mitogens and stellation agents, to study the early nuclear events which accompany the induction of glial proliferation and/or differentiation. Tetradecanoyl phorbol acetate (TPA), epidermal growth factor, and fibroblast growth factor, three mitogens for astrocytes, stimulated marked, rapid, and transient increases in TIS gene mRNAS.

HeLa cell mutants resistant to epidermal growth factor ricin A-chain conjugate.

Epidermal growth factor (EGF) was linked to the toxic A chain of ricin toxin (RTA) to produce an EGF-receptor-specific cytotoxic agent, EGF-RTA. Three EGF-RTA-resistant mutants of the human HeLa cell line were selected. These mutant cell lines are 10-fold to more than 100-fold more resistant to EGF-RTA when compared to HeLa cells.

An epidermal growth factor-ricin A chain (EGF-RTA)-resistant mutant and an epidermal growth factor-Pseudomonas endotoxin (EGF-PE)-resistant mutant have distinct phenotypes.

H2Oe12 is a mutant HeLa cell line selected for resistance to the toxicity of a chimeric protein conjugate composed of epidermal growth factor (EGF) and the toxic A chain of ricin (RTA). ET-28 is a mutant KB cell line selected for resistance to the toxicity of a chimeric protein conjugate composed of EGF and Pseudomonas exotoxin (PE). In this report we describe the presence or absence, in these mutants, of cross-resistance to the two toxic conjugates and the effects of ammonium chloride, leupeptin, and adenovirus cotreatments on toxin efficacies.

Induction of tumor promotor-inducible genes in murine 3T3 cell lines and tetradecanoyl phorbol acetate-nonproliferative 3T3 variants can occur through protein kinase C-dependent and -independent pathways.

We isolated a group of genes that are rapidly and transiently induced in 3T3 cells by tetradecanoyl phorbol acetate (TPA). These genes are called TIS genes (for TPA-inducible sequences). Epidermal growth factor (EGF), fibroblast growth factor (FGF), and TPA activated TIS gene expression with similar induction kinetics.

Tumorigenic methylcholanthrene transformants of C3H/10T1/2 cells have a common nucleotide alteration in the c-Ki-ras gene.

The polymerase chain reaction was used to amplify DNA surrounding the codon 12 region of the c-Ki-ras gene from C3H/10T1/2 cells and from a number of 3-methylcholanthrene (MCA)-transformed derivatives of these cells. Sequence analysis demonstrated that tumorigenic MCAC116/39 cells, known by DNA-mediated transfection to contain an activated c-Ki-ras oncogene, had a G----T transversion in the first position of codon 12 of this gene, resulting in a Gly12----Cys mutation. A combination of polymerase chain-reaction amplification and oligonucleotide hybridization demonstrated that three additional tumorigenic MCA transformants of C3H/10T1/2 cells had an identical mutation in the c-Ki-ras gene.

Nucleotide sequence of a cDNA encoding TIS11, a message induced in Swiss 3T3 cells by the tumor promoter tetradecanoyl phorbol acetate.

We report here the nucleic acid sequence and deduced amino acid sequence of a cDNA for TIS11, a gene induced in 3T3 cells by tetradecanoyl phorbol acetate.

Detection and quantification of S-100 protein in ocular tissues and fluids from patients with intraocular melanoma.

S-100 protein is a 21,000 dalton acidic calcium-binding protein present in ocular melanomas and some normal ocular tissues. Ocular fluids and extracts of ocular tumours were examined by a sensitive radioimmunoassay that could detect less than 5 ng of S-100 protein in minute volumes of fluid. Three ocular melanoma biopsy specimens had S-100 protein at levels between 25 and 1300 ng/ml, comparable to that found in a cutaneous melanoma biopsy specimen (1000 ng/ml).

Tumor promoter-inducible genes are differentially expressed in the developing mouse.

TIS genes are rapidly and transiently induced by tetradecanoyl phorbol acetate in 3T3 cells. We analyzed the developmental appearance of a number of the TIS genes to determine whether, in a normal physiological context, these genes have common or distinct mechanisms of regulation. Each TIS gene has a distinct tissue specificity and/or developmental profile.

A continuum of transformed phenotypes in C3H/10T1/2 derivatives.

Two-stage transformants have been isolated from C3H/10T1/2 cells exposed to UV-irradiation followed by treatment with TPA. These UV-TDTx cells form foci in co-culture with C3H/10T1/2 cells only in the presence of TPA. In the absence of TPA, UV-TDTx cells are indistinguishable from control C3H/10T1/2 cells in co-cultures.

A phorbol ester-nonproliferative variant of Swiss 3T3 cells is deficient in Na+K+Cl- cotransport activity.

The identity of the genetic defect(s) in Swiss 3T3 TNR-2 and TNR-9 that confers nonresponsiveness to the proliferative effect of 12-0-tetradecanoylphorbol-13-acetate (TPA) is not known. In BALB/c 3T3 cells, loss (via mutation) of a specific membrane ion transport system, the furosemide-sensitive Na+K+Cl- cotransporter, is associated with decreased responsiveness to TPA. In this study, the transport properties of parental Swiss 3T3 cells and the TPA-nonresponsive lines TNR-2 and TNR-9 were determined in the presence and absence of TPA.

Butyrate-induced changes in nuclease sensitivity of chromatin cannot be correlated with transcriptional activation.

We examined in the H4IIE rat hepatoma cell line the relationship between butyrate-induced changes in the nuclease sensitivity of chromatin and changes in transcriptional activity of specific genes. The butyrate-inducible metallothionein I (MT-I) gene underwent a dramatic increase in DNase I sensitivity after 3 h of butyrate treatment. However, genes not transcribed in H4IIE cells underwent the same changes in DNase I sensitivity.

Metal-dependent binding of a factor in vivo to the metal-responsive elements of the metallothionein 1 gene promoter.

Using the technique of genomic footprinting, we demonstrate cadmium-inducible protection from dimethyl sulfate (DMS) modification of guanine residues in vivo in five metal-responsive elements (MREs) in the promoter of the rat metallothionein 1 (MT-1) gene. We also identify a site of extreme DMS hyperreactivity which, like the MRE protection, occurs only after metal ion induction. With this hyperreactive site as an indicator, we can measure the kinetics of induction and deinduction.

Colony size, cell density and nature of the tumor promoter are critical variables in expression of a transformed phenotype (focus formation) in co-cultures of UV-TDTx and C3H10T1/2 cells.

UV-TDTx cells are cloned from foci arising after C3H10T1/2 cells are sequentially exposed to u.v. irradiation followed by tetradecanoylphorbol acetate (TPA).

Reactivity of neoplastic cells of hairy cell leukemia with antisera to S-100 protein.

Rabbit antibodies to bovine S-100 protein were tested by immunoperoxidase technics against fresh hairy cell leukemia (HCL) cells obtained from nine patients (peripheral blood in six and spleen in three), as well as lymphoblastoid cell lines derived from three patients with HCL. Peripheral mononuclear cells from three normal persons and two patients with chronic lymphocytic leukemia (CLL) and cells from two melanoma lines were used as controls. The melanoma cell lines, cell lines derived from patients with HCL, and fresh HCL cells displayed cytoplasmic and nuclear positivity after exposure to anti-S-100 protein sera.

Toxicity of ligand and antibody-directed ricin A-chain conjugates recognizing the epidermal growth factor receptor.

Approximately equal amounts of 125I-mAb 225 (a monoclonal antibody recognizing the human epidermal growth factor receptor) and 125I-labeled epidermal growth factor (125I-EGF) were bound by HeLa cells. However, these two EGF receptor binding moieties had different fates after binding. Sixty percent of cell-associated 125I-EGF was internalized.

Zinc and copper in milk and tissues of nursing lethal milk mutant mice.

Zinc concentration was lower in liver of suckling 1-d-old lethal milk (lm/lm) mutant mice than in wild-type pups, in accordance with the hypothesis of milk-induced zinc deficiency previously proposed to underlie this mutation. Despite the initial deficiency, by 3 d of age suckling lm/lm pups exhibited higher levels of hepatic zinc than did lm/lm-nursed wild-type pups. Intestinal zinc and copper concentrations were normal in 1-d-old lm/lm pups, but by 3 d of age were also higher in lm/lm pups than in wild-type pups foster-nursed on lm/lm dams.

Metallothionein synthesis and mRNA levels in cultured hepatocytes from lethal milk mutant mice.

Primary hepatocyte cultures from adult lethal milk (lm) mutant mice displayed approximately a 2.5-fold increased metallothionein (MT) synthesis compared to wild-type cells in uninduced as well as zinc-treated cells, over a 0- to 160-microM zinc dose-response range. Mutant hepatocytes also exhibited a greater level of MT-1 mRNA in both untreated and zinc-treated cultures compared to wild-type.

Induction of transiently expressed genes in PC-12 pheochromocytoma cells.

Rat PC-12 pheochromocytoma cells, in response to nerve growth factor (NGF), stop proliferating and differentiate into cells resembling sympathetic neurons. This model of cell differentiation was used to investigate the expression of a previously isolated collection of mitogen-induced primary response sequences cloned from murine 3T3 cells; the TIS (tetradecanoyl phorbol acetate-induced sequences) genes (Lim et al., 1987).