Occurrence of epidermal growth factor receptors in human adnexal tumors and their prognostic value in advanced ovarian carcinomas

Bauknecht, Thomas; Runge, Michael C.; Schwall, Michael; Pfleiderer, Albrecht

doi:10.1016/0090-8258(88)90209-0

Cited by 68 publications

(28 citation statements)

References 13 publications

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“…It has been noted by Bauknecht et al (1988) that a response to chemotherapy occurred in 50% of EGFR positive cancers with a mean survival time of 28 months while the response rate in EGFR negative ovarian cancers was 12% with a mean survival of 16 months. Our results have not yet been analysed in this way as it was felt that the time interval was not sufficiently long.…”

Section: Resultsmentioning

confidence: 99%

Epidermal growth factor receptors (EGFR) in human ovarian cancer

Owens

Cc²,

Brown³

et al. 1991

Br J Cancer

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Preparation of membrane pellet Tumour specimens removed from -70'C were allowed to thaw on ice, while those from sucrose/glycerol were re-hydrated in homogenising buffer (see below). The tumour specimen was dried with tissue paper to remove any excess water or buffer. Specimens were washed in ice cold saline. The tumour was bisected and two separate samples of tumour were cut (2-3 mm minimum) from either half, one piece placed in formal saline for pathological analysis and the other in sucrose/glycerol buffer for later immunohistochemical analysis (to be reported later). The remainder of the tumour was used for the biochemical assay. Fresh homogenising buffer was prepared (20 mM Hepes, 2 mM EDTA, 0.5 mM PMSF to pH 7.4). Tumour sections were then cut into small 1 mm blocks and weighed (usually 1 gram), then placed in a centrifuge tube to which was added 5 ml g-' (wet weight) of ice cold homogenising buffer. Tumour was homogenised on ice with an ultra turrax (Janke and Kunkel) with 2 x 15 s bursts at maximum speed but allowing the homogenate to cool between bursts. The homogenate was centrifuged at 1,000 g for 10 min. The resulting supernatant was centrifuged at 12,000 g for 1 h. The nuclear pellet was resuspended in homogenising buffer and stored at -20C until required for DNA analysis (Modified Burton). The pellet from the high speed spin was resuspended in 1-2 ml of radioimmunoassay buffer (RIA buffer: 0.2 M Na2HPO4, 0.2 M NaH2PO4, 0.1% sodium azide, 0.15 M sodium chloride, 0.1 M EDTA and 0.5% bovine serum albumin to pH 7.4) depending on the initial wet weight (1 ml 500 mg-') and stored at -20C in 1.5 ml polystyrene tubes (Eppendorfs). Prior to storage each suspension was submitted to glass teflon homogenisation to ensure an even suspension.

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Section: Resultsmentioning

confidence: 99%

Epidermal growth factor receptors (EGFR) in human ovarian cancer

Owens

Cc²,

Brown³

et al. 1991

Br J Cancer

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“…At present, although EGFR expression has been widely demonstrated in ovarian cancer (Bauknecht et al, 1988;Battaglia et al, 1989;Berchuck et al, 1991;Stewart et al, 1992;van der Burg et al, 1993), few studies have investigated the prognostic significance of EGFR (Bauknecht et al, 1988;Berchuck et al, 1991;van der Burg et al, 1993). Some evidence does, however, demonstrate that high EGFR levels may be a negative prognostic indicator in many tumour types (Sainsbury et al, 1987;Neal et al, 1991;Maurizi et al, 1992;Scambia et al, 1994).…”

mentioning

confidence: 99%

Epidermal growth factor, oestrogen and progesterone receptor expression in primary ovarian cancer: correlation with clinical outcome and response to chemotherapy

Scambia

Benedetti‐Panici²,

Ferrandina³

et al. 1995

Br J Cancer

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Summary The expression of epidermal growth factor receptor (EGFR), oestrogen receptor (ER) and progesterone receptor (PR) was assayed by a radioreceptor method in 117 primary ovarian cancers. EGFR was not significantly related to any of the clinicopathological parameters examined. In patients with stage II-IV disease who underwent second-look surgery after primary chemotherapy, a significant correlation between high EGFR levels and poor response to chemotherapy was demonstrated (P = 0.031). Moreover, post-operative residual tumour showed an independent role in predicting chemotherapy response (P = 0.0007) and EGFR status showed a borderline significance (P = 0.052) in the multivariate analysis. No correlation between steroid hormone receptors and clinicopathological parameters was observed. Whereas a significant relationship was shown between EGFR positivity and a shorter overall survival (OS) (P = 0.0022) and progression-free survival (PFS) (P = Neal et al., 1991;Maurizi et al., 1992; Scambia et al., 1994). Moreover, in breast cancer EGFR expression seems to be a feature of hormone-independent aggressive clinical behaviour (Klijn et al., 1992). Our previous study The membrane fraction and cytosol were prepared as described elsewhere . Briefly, tumour specimens were finely minced and homogenised in five volumes of ice-cold buffer consisting of 25 mM Tris, 1.5 mM EDTA, 5 mM sodium azide, 20% glycerol (TENG) plus 0.1 % monothioglycerol, by applying several intermittent bursts of an Ultra-Turrax homogeniser. The crude homogenate was centrifuged at 7000 g for 20 min at 0°C in order to separate nuclear fraction from the cytosol fraction. The supernatants were then centrifuged at 105 000 g for 75 min at 0°C, obtaining the membrane pellet and the cytosolic fraction.

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“…High levels of EGFR expression in multiple tumor types is therefore related to an increased probability of tumor recurrence and poor patient survival. The presence of EGFR in ovarian cancer has been well demonstrated using various methods such as ligand binding, immunohistochemistry, or Northern blot analysis (Bauknecht et al 1988, Battaglia et al 1989, Berchuck et al 1991, Morishige et al 1991, Owens et al 1991, HenzenLogmans et al 1992, Bauknecht et al 1993. EGFR is also frequently amplified and/or overexpressed when compared with normal OSE, and transfection with an antisense construct of EGFR into human ovarian cancer cell lines suppressed the malignant phenotype, cellular proliferation and tumorigenicity of these cells, suggesting its prognostic importance (Berns et al 1992, Brader et al 1998, Alper et al 2000.…”

Section: Introductionmentioning

confidence: 99%

Gonadotropins upregulate the epidermal growth factor receptor through activation of mitogen-activated protein kinases and phosphatidyl-inositol-3-kinase in human ovarian surface epithelial cells

Choi¹

2005

Endocrine Related Cancer

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Gonadotropins, follicle-stimulating hormone (FSH) and luteinizing hormone (LH) have been implicated as probable risk factors in epithelial ovarian carcinomas, most of which are derived from ovarian surface epithelium (OSE). Since epidermal growth factor (EGF) increases the growth of ovarian surface epithelial cells, we determined the effect of gonadotropins on the expression of epidermal growth factor receptor (EGFR). We investigated the basal levels of EGFR mRNA and protein, and the mechanisms involved in the regulation of EGFR at the transcriptional and translational levels by FSH and LH. The immortalized OSE cell lines (IOSE) derived from normal OSE cells by transfecting SV40 T-antigen (IOSE-80 and IOSE-80PC, a post-crisis line) and ovarian cancer cell lines were employed. A significantly lower level of EGFR was observed in both IOSE-80 and IOSE-80PC cells when compared with the ovarian cancer cell lines, OVCAR-3 and SKOV-3. Treatment of IOSE-80PC cells with FSH and LH (10 -7 and 10 -6 g/ml) resulted in a significant increase in EGFR mRNA at 24 h and EGFR protein at 48 h, whereas the treatment with gonadotropins for 24-48 h induced a mild increase in EGFR in OVCAR-3, but not in SKOV-3 cells. In addition, IOSE-80PC cells treated with gonadotropins and EGF (10 nM) exhibited an additive stimulation of mitogenesis. Further, FSH and LH significantly increased activities of various kinases at 5-10 min, and pre-treatments with LY294002 (an inhibitor of PI3K) or PD98059 (an inhibitor of ERK1/2) partially blocked the gonadotropin-induced up-regulation of EGFR in IOSE-80PC cells. We investigated whether the effect of gonadotropins on EGFR mRNA levels is induced by increased transcription and/or by altered mRNA stability. Treatment of IOSE-80PC cells with FSH (10 -7 and 10 -6 g/ml) significantly enhanced the activity of the EGFR promoter (120 and 140% increase, respectively) at 24 h, and treatment with LH (10 -7 g/ml) for 24 h induced an increase in the activity of EGFR promoter (30%) in these cells. On the other hand, LH resulted in a significant increase in EGFR mRNA stability in the decay curves. Taken together, these results suggest that the effect of gonadotropins on the expression of EGFR may affect cell growth via ERK-1/-2 and PI3K pathways in preneoplastic ovarian surface epithelial cells, and that FSH and LH increase EGFR mRNA by different mechanisms. The former increased EGFR gene transcription essentially, whereas the latter mainly enhanced EGFR mRNA stability.

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Occurrence of epidermal growth factor receptors in human adnexal tumors and their prognostic value in advanced ovarian carcinomas

Cited by 68 publications

References 13 publications

Epidermal growth factor receptors (EGFR) in human ovarian cancer

Epidermal growth factor receptors (EGFR) in human ovarian cancer

Epidermal growth factor, oestrogen and progesterone receptor expression in primary ovarian cancer: correlation with clinical outcome and response to chemotherapy

Gonadotropins upregulate the epidermal growth factor receptor through activation of mitogen-activated protein kinases and phosphatidyl-inositol-3-kinase in human ovarian surface epithelial cells

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