Comparison of nuclear proteins from DMBA-induced mammary tumors and lactating mammary glands by polyacrylamide gel electrophoresis

Dinh, Bao-Linh; Bourget, Lucie

doi:10.1007/bf01807648

Cited by 5 publications

(1 citation statement)

References 23 publications

(17 reference statements)

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“…The method we had previously reported for the isolation of tumor cell nuclei from experimental mammary tumors was used with slight modifications (Dinh and Bourget, 1985). For each nuclear preparation, 2-4 breast tumor specimens totalling about 3 g of tissue were needed.…”

Section: Isolation and Purijication Of Nucleimentioning

confidence: 99%

Generation of monoclonal antibodies reactive with nuclear proteins of human primary breast tumors

Dinh

Tremblay

Paradis

et al. 1988

Intl Journal of Cancer

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Nuclear proteins were extracted from purified nuclei of human primary breast tumors (BrT) and bladder tumors and of human normal breast, kidney and lymphocytes by enzymatic treatment. SDS-Polyacrylamide gel electrophoresis of nuclear proteins from breast tumors showed different bands in the molecular weight zones from 25 to 220 kDa which were absent or present only as traces in normal breast tissue. Murine monoclonal antibodies (MAbs) have been produced using nuclear extracts of human primary breast tumors as immunogens. Approximately 2,000 hybridomas were generated from 5 hybridizations. According to their reactivity to BrT nuclear extracts and mammary carcinoma cell line MCF-7, seven hybridomas were selected and cloned. They were further characterized with histological immunoperoxidase assays of formaldehyde-fixed BrT paraffin tissue sections. MAb 6A3 particularly gave strong nuclear staining with all BrT specimens while MAb 1D8 showed both nuclear and cytoplasmic staining with only some of them. Specimens from mammoplasty did not react with these MAbs. Immunoblotting of BrT nuclear extracts as developed with MAbs 6A3 and 1D8 revealed major protein bands with molecular weight of 120 and 130 kDa. The potential use of these MAb-defined BrT-related nuclear proteins as markers for human breast cancer was suggested.

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Section: Isolation and Purijication Of Nucleimentioning

confidence: 99%

Generation of monoclonal antibodies reactive with nuclear proteins of human primary breast tumors

Dinh

Tremblay

Paradis

et al. 1988

Intl Journal of Cancer

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Specific polypeptide differences in normal versus malignant breast tissue by two‐dimensional electrophoresis

Wirth¹

1989

Electrophoresis

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High resolution two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) in combination with computer-assisted densitometry was used to analyze 800-1000 silver stained postmitochrondrial and 600-800 cytosolic polypeptides extracted from malignant and nonmalignant human breast tissues. The 2D-PAGE patterns of polypeptides from malignant and normal tissues were similar, although both qualitative and quantitative polypeptide differences were noted. Six cytosolic polypeptides (pI/molecular mass x 10(-3), 5.20/80, 5.75/43, 6.20/40, 5.43/35, 5.46/34.5, and 5.50/34 were detected exclusively in malignant tissues. One constitutive polypeptide, p52 (7.25/52), was not detected in tumor samples. Marked quantitative differences in spot density were noted in polypeptides localized mainly in the molecular weight ranges of 22-40 kDa and pI of 5.65-7.00. An overall increase in polypeptide expression was noted in this region of 2D-PAGE gels of malignant tissues as compared to normal. Twenty-two acidic and 19 polypeptides separated under nonequilibrium isoelectric focusing conditions were significantly increased in tumor samples while one polypeptide was decreased. One polypeptide, p24 (6.15/24), was expressed in greatest concentrations in tumors which also expressed the greatest estrogen receptor content. Expression of p24 was markedly reduced in normal tissue and in malignant tissues expressing low levels of estrogen and progesterone receptors. No significant differences in the expression of the Yb and Ya subunits of glutathione-S-transferases (GST)-A, -B and ligandin were observed between normal and malignant breast tissue. None of the Yp subunits of the placental isoform of GST were detected in either normal or malignant breast tissues.

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Molecular and functional diversity of non-histone protein fraction NHCP1 from hamster Kirkman-Robbins hepatoma and liver

et al. 1986

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Non-histone protein fraction NHCP1 of micrococcal nuclease-sensitive and nuclease-resistant chromatin from Kirkman-Robbins hepatoma and hamster liver was studied by two-dimensional electrophoresis followed by Coomassie and silver staining and by microcomplement fixation technique in the presence of antibodies elicited against NHCP1 of both tissues. Apart from many common spots several tissue specific components associated with either nuclease-sensitive or nuclease-resistant chromatin were found. The presence of tissue specific components among NHCP1 from hepatoma and liver was confirmed by immunological analysis. It was stated that these components are exclusively localized in nuclease-resistant part of chromatin from neoplastic and normal tissues thus suggesting their structural function.

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Comparison of nuclear proteins from DMBA-induced mammary tumors and lactating mammary glands by polyacrylamide gel electrophoresis

Cited by 5 publications

References 23 publications

Generation of monoclonal antibodies reactive with nuclear proteins of human primary breast tumors

Generation of monoclonal antibodies reactive with nuclear proteins of human primary breast tumors

Specific polypeptide differences in normal versus malignant breast tissue by two‐dimensional electrophoresis

Molecular and functional diversity of non-histone protein fraction NHCP1 from hamster Kirkman-Robbins hepatoma and liver

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