Primary Cultures of Normal and Tumoral Human Ovarian Epithelium: A Powerful Tool for Basic Molecular Studies

Lounis, H.; Provencher, Diane; Godbout, Charles; Fink, Dorit; Milot, Martin; Mes-Masson, Anne-Marie

doi:10.1006/excr.1994.1346

Cited by 55 publications

(86 citation statements)

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“…The success of our approach may in part be explained by the choice of the model system, which does not rely on primary undissected tumour tissue that may contain several cell types. In particular, we have previously demonstrated that even short-term passage of primary cultures results in an enriched homogeneous cell population (Lounis et al, 1994). The absence of non-malignant contaminating cells in our samples has probably allowed a strict selection of genes specifically expressed in epithelial cells.…”

Section: Discussionmentioning

confidence: 86%

“…Normal tissues were obtained from tumour-free participants that have undergone oopherectomies. Primary cell cultures from NOSE and EOC samples were established as described (Kruk et al, 1990;Lounis et al, 1994) and used for microarray analysis. Cells in primary culture were maintained in OSE media consisting of 50 : 50 medium 199:105 (Sigma, St Louise, MO, USA) supplemented with 10% fetal bovine serum, 2.5 mg ml À1 amphotericin B and 50 mg ml À1 gentamicin (Kruk et al, 1990).…”

Section: Patients Cell Culture and Clinical Materialsmentioning

confidence: 99%

“…As normal epithelial cells are generally quiescent, differential gene expression observed between tissues of NOSE and EOC may reflect differences not necessarily related to oncogenic transformation. In ovarian cancer, primary cultures derived from NOSE and tumour tissue have been described (Lounis et al, 1994). In the case of EOC tissues, primary cultures have the advantage of being relatively free of nontransformed cell population.…”

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confidence: 99%

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Gene expression profiling of primary cultures of ovarian epithelial cells identifies novel molecular classifiers of ovarian cancer

et al. 2006

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In order to elucidate the biological variance between normal ovarian surface epithelial (NOSE) and epithelial ovarian cancer (EOC) cells, and to build a molecular classifier to discover new markers distinguishing these cells, we analysed gene expression patterns of 65 primary cultures of these tissues by oligonucleotide microarray. Unsupervised clustering highlights three subgroups of tumours: low malignant potential tumours, invasive solid tumours and tumour cells derived from ascites. We selected 18 genes with expression profiles that enable the distinction of NOSE from these three groups of EOC with 92% accuracy. Validation using an independent published data set derived from tissues or primary cultures confirmed a high accuracy (87 -96%). The distinctive expression pattern of a subset of genes was validated by quantitative reverse transcription -PCR. An ovarian-specific tissue array representing tissues from NOSE and EOC samples of various subtypes and grades was used to further assess the protein expression patterns of two differentially expressed genes (Msln and BMP-2) by immunohistochemistry. This study highlights the relevance of using primary cultures of epithelial ovarian cells as a model system for gene profiling studies and demonstrates that the statistical analysis of gene expression profiling is a useful approach for selecting novel molecular tumour markers.

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Section: Discussionmentioning

confidence: 86%

Section: Patients Cell Culture and Clinical Materialsmentioning

confidence: 99%

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confidence: 99%

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Gene expression profiling of primary cultures of ovarian epithelial cells identifies novel molecular classifiers of ovarian cancer

et al. 2006

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“…Normal controls were defined as tumor-free patients. Primary cell cultures from NOSE and EOC samples were established as described 15,16 and used for microarray analysis. Cells in primary culture were maintained for a maximum of 17 passages in OSE media consisting of 50:50 medium 199:105 (Sigma) supplemented with 10% fetal bovine serum (FBS), 2.5lg/ ml amphotericin B and 50 lg/ml gentamicin.…”

Section: Clinical Samplesmentioning

confidence: 99%

From gene profiling to diagnostic markers: IL‐18 and FGF‐2 complement CA125 as serum‐based markers in epithelial ovarian cancer

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Ouellet

Madore

et al. 2005

Intl Journal of Cancer

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We used an oligonucleotide-based DNA microarray to identify potential markers in 39 primary cultures of ovarian cancer specimens compared with 11 primary cultures of normal ovarian epithelia. Differential gene expression of IL-18 and FGF-2 was validated on a subset of samples by quantitative PCR and by IHC, using an independent tissue array of 90 cores of 20 normal ovarian surface epithelia and 70 EOCs representing different grades and pathologies of ovarian disease. We further compared, by ELISA, these two markers with CA125 in sera from 25 cancer-free and 47 ovarian cancer patients. IL-18 and FGF-2 proteins were significantly elevated in tumor tissues (p<0.04) and sera (p<0.05) from patients with ovarian cancer. In combination, the three markers (IL-18, FGF-2, and CA125) showed similar sensitivity in scoring for ovarian cancer (35/45 patients) compared to that of CA125 alone (37/45) and significantly improved the specificity of detection (20/25 patients) compared to each marker individually (15/25 for CA125; 18/25 FGF-2; 16/25 for IL-18). In conclusion we show that a combination of the three serum markers (IL-18, FGF-2 and CA125) is associated with EOC, with higher specificity than CA125 alone. Prospective studies with a large cohort of susceptible ovarian cancer patients will be required to expand these findings. ' 2005 Wiley-Liss, Inc.

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“…DNA was extracted from solid tumors, snap frozen in liquid nitrogen at the time of resection, and from peripheral lymphocytes according to the method described previously (Lounis et al, 1994).…”

Section: Dna Extractionmentioning

confidence: 99%

Allelotyping defines minimal imbalance at chromosomal region 17q25 in non-serous epithelial ovarian cancers

et al. 2000

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Allelic deletions of multiple chromosome 17q loci in sporadic ovarian cancer of epithelial origin suggest that inactivation of tumor suppressor gene(s) in these regions may be important for ovarian tumorigenesis. To further de®ne the pattern of allelic imbalance in epithelial ovarian tumors of di erent histologies, a PCR-based assay was used to assess loss of heterozygosity (LOH) of polymorphic markers representative of TP53, BRCA1, NME1 and GH1, and region 17q23-25. LOH was observed for at least one marker in 68% of malignant tumors (n=60) and in 18% tumors of borderline malignancy (n=11), but not in benign tumors (n=5). The highest frequency of LOH in malignant tumors (64%) was observed with D17S801 on 17q25. Ten of 39 malignant ovarian tumors displaying LOH of at least one 17q marker, displayed a LOH pattern enabling the determination of a minimal region of overlapping deletion de®ned by D17S795 and D17S801. One borderline tumor also displayed an interstitial LOH pattern that overlapped this 17q25 minimal region of deletion. The histologies of malignant tumors displaying a pattern indicative of interstitial 17q deletions were of the endometrioid, clear cell and mucinous epithelial types. As the minimal region of overlap de®ned by these tumors overlap regions deleted in malignant tumors of all histologic types, and in a tumor of borderline malignancy, the 17q25-tumor suppressor may be implicated in the development of all types of epithelial ovarian tumors.

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Primary Cultures of Normal and Tumoral Human Ovarian Epithelium: A Powerful Tool for Basic Molecular Studies

Cited by 55 publications

References 0 publications

Gene expression profiling of primary cultures of ovarian epithelial cells identifies novel molecular classifiers of ovarian cancer

Gene expression profiling of primary cultures of ovarian epithelial cells identifies novel molecular classifiers of ovarian cancer

From gene profiling to diagnostic markers: IL‐18 and FGF‐2 complement CA125 as serum‐based markers in epithelial ovarian cancer

Allelotyping defines minimal imbalance at chromosomal region 17q25 in non-serous epithelial ovarian cancers

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