Cervical brush biopsy specimens suitable for DNA and oncoprotein analysis using flow cytometry.

Elias-Jones, J; Hendy-Ibbs, P; Cox, Holly D.; Gi, Evan; Watson, J.

doi:10.1136/jcp.39.5.577

Cited by 12 publications

(1 citation statement)

References 18 publications

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“…We have recently developed methods using the Cyto-Brush (Medscand, Malmo, Sweden) to collect cervical epithelium for this assay on a routine bases from colposcopy clinics (Elias-Jones et al, 1986). The cell yield was good and was not contaminated with stromal cells which is another potential artefact source in punch biopsy material.…”

Section: Dna Analysesmentioning

confidence: 99%

Flow cytometric quantitation of DNA and c-myc oncoprotein in archival biopsies of uterine cervix neoplasia

Hendy-Ibbs¹,

Cox²,

Gi³

et al. 1987

Br J Cancer

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Summary The c-myc nuclear associated oncoprotein has been quantitated simultaneously with DNA in nuclei extracted from archival biopsies of uterine cervix neoplasia. The oncoprotein and DNA were measured fluorimetrically in a flow cytometer using a mouse monoclonal antibody (MYC I-6E10) and propidium iodide. Normal biopsies exhibited higher oncoprotein levels than carcinomas (P<0.00001). Furthermore, the maximum fluorescence signal in the normal tissue occurred at a lower antibody concentration compared with tumour tissue. There was no correlation between oncoprotein levels and histological grade, stage of disease, age of the patients or prognosis in the carcinomas. Aneuploidy, defined as a distinct second peak separate from the diploid distribution, was not a significant feature. The c-myc oncoprotein nuclear content does not appear to be a prognostic indicator in carcinoma of the cervix from the results of these studies but there is clearly diagnostic potential, particularly for automated analysis of cervical screening.Oncogcincs are associated with proliferation control. The csis gene encodes a subunit of platelet derived growth factor (Doolittle et al., 1983; Waterfield et al., 1983). v-erb B and cfms respectively encode the intracellular domain of epidermal growth factor receptor (Downward et al., 1984) and the transmembrane receptor for the macrophage colony stimulating factor, CSF 1, (Scherr et al., 1985). Expression of the c-myc gene is associated with the transition from a quiescent to a stimulated state (Kelly et al., 1983(Kelly et al., , 1984Makino et al., 1984;Greenberg & Ziff, 1984;Hann et al., 1985;Rabbitts et al., 1985).A series of mouse monoclonal antibodies which recognise the c-myc nuclear associated oncoprotein, p62c-mYc, (Evan & Hancock, 1985) have been developed . One of these antibodies has been used for histological localization of p62c-myc in both testicular cancer and in colonic neoplasia using immunocytochemical techniques. Quantitative methods have now been developed to assay nuclear associated p62cmvc in individual nuclei extracted from archival biopsies using flow cytometry . These methods have been used to analyse data in testicular cancer and colonic neoplasia (Watson et al., submitted). In this paper we examine the p62c-mYc nuclear content in normal and neoplastic cervical biopsies. Patients and methods PatientsA total of 127 patients attending the Weston Park Hospital Radiotherapy Department, Sheffield between 1971 and 1978 were included in the study. The only criteria of entry were that sufficient wax embedded punch biopsy material was available for the assay and that there were complete followup data. Sixty-four biopsies were from invasive carcinoma and 29 were from normal cervix. The remaining patients had cervical intraepithelial neoplasia (CIN) disease where 11, 9 and 14 were grades I, II and III respectively.Anti-p62c-mYc antibody Full details of the methods for production of the antip62c-mYc antibody are published elsewhere . Briefly, synthetic peptides were constructe...

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Section: Dna Analysesmentioning

confidence: 99%