2.50
Hdl Handle:
http://hdl.handle.net/10541/78134
Title:
TSLC1 gene silencing in cervical cancer cell lines and cervical neoplasia.
Authors:
Steenbergen, R D; Kramer, D; Braakhuis, B J; Stern, Peter L; Verheijen, René H; Meijer, C J; Snijders, P J
Abstract:
BACKGROUND: Cervical carcinogenesis is initiated by infection with high-risk (i.e., carcinogenic) human papillomavirus (HPV) types. The subsequent progression from premalignant cervical intraepithelial neoplasia (CIN) to invasive cancer is driven by both genetic and epigenetic processes. We assessed the role of the gene encoding the adhesion molecule tumor suppressor in lung cancer 1 (TSLC1) in this progression. METHODS: We analyzed TSLC1 gene expression by real-time quantitative reverse transcription-polymerase chain reaction, promoter methylation by sodium bisulfite genomic DNA sequencing, and allelic loss by microsatellite analysis in primary keratinocytes, in four non-tumorigenic HPV-immortalized human keratinocyte cell lines, and in 11 human cervical cancer cell lines that were positive for a high-risk HPV DNA type and in normal cervical epithelial cells. We transfected cervical cancer SiHa cells that did not express TSLC1 mRNA with an expression vector containing the TSLC1 complementary DNA (cDNA) or an empty vector and analyzed transfectants for anchorage-independent growth and tumorigenicity in nude mice. We also examined TSLC1 promoter methylation in premalignant cervical lesions and in cervical carcinomas and smears. All statistical tests were two-sided. RESULTS: TSLC1 mRNA was strongly reduced, relative to levels in primary keratinocytes, or absent in 10 (91%) of 11 cervical carcinoma cell lines but in none (0%) of the four HPV-immortalized cell lines (difference = 91%, 95% confidence interval [CI] = 74% to 100%; P =.004). The TSLC1 promoter was hypermethylated, relative to normal foreskin and cervical epithelial cells, in nine (82%) of the 11 cervical carcinoma cell lines but in none (0%) of the four HPV-immortalized cell lines (difference = 82%, 95 CI = 59% to 100%; P =.01). Seven (88%, 95% CI = 47% to 100%) of the eight SiHa/TSLC1 transfectants displayed a marked reduction in anchorage-independent growth (i.e., 0-100 colonies per 5000 cells) compared with none of the four (0%, 95% CI = 0% to 60%) SiHa transfectants bearing the empty vector (i.e., SiHa/hygro transfectants; difference = 88%, 95% CI = 65% to 100%; P =.01) or untransfected SiHa cells. All seven mice (100%, 95% CI = 59% to 100%) injected with untransfected SiHa cells or SiHa/hygro transfectants displayed tumors of at least 50 mm(3) by 2-6 weeks after injection compared with none of eight mice (0%, 95% CI = 0% to 37%) injected with the SiHa/TSLC1 transfectants (difference = 100%, 95% CI = 68% to 100%; P<.001). We detected TSLC1 promoter hypermethylation in seven (35%, 95% CI = 15% to 59%) of 20 high-grade CIN lesions (i.e., CIN II and III) and in 30 (58%, 95% CI = 43% to 71%) of 52 cervical squamous cell carcinomas compared with none (0%, 95% CI = 0% to 34%) of nine normal cervical epithelial biopsy samples and none (0%, 95% CI = 0% to 22%) of 12 CIN I lesions (P<.001 for cervical squamous cell cancer versus normal epithelial biopsy samples plus CIN I lesions). CONCLUSIONS: TSLC1 gene silencing via promoter hypermethylation is a frequent event in the progression from high-risk HPV-containing, high-grade CIN lesions to invasive cervical cancer.
Affiliation:
Department of Pathology, Unit of Molecular Pathology, Vrije Universiteit Medical Center, Amsterdam, The Netherlands. r.steenbergen@vumc.nl
Citation:
TSLC1 gene silencing in cervical cancer cell lines and cervical neoplasia. 2004, 96 (4):294-305 J. Natl. Cancer Inst.
Journal:
Journal of the National Cancer Institute
Issue Date:
18-Feb-2004
URI:
http://hdl.handle.net/10541/78134
DOI:
10.1093/jnci/djh031
PubMed ID:
14970278
Type:
Article
Language:
en
ISSN:
1460-2105
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorSteenbergen, R Den
dc.contributor.authorKramer, Den
dc.contributor.authorBraakhuis, B Jen
dc.contributor.authorStern, Peter Len
dc.contributor.authorVerheijen, René Hen
dc.contributor.authorMeijer, C Jen
dc.contributor.authorSnijders, P Jen
dc.date.accessioned2009-08-21T10:17:05Zen
dc.date.available2009-08-21T10:17:05Zen
dc.date.issued2004-02-18en
dc.identifier.citationTSLC1 gene silencing in cervical cancer cell lines and cervical neoplasia. 2004, 96 (4):294-305 J. Natl. Cancer Inst.en
dc.identifier.issn1460-2105en
dc.identifier.pmid14970278en
dc.identifier.doi10.1093/jnci/djh031en
dc.identifier.urihttp://hdl.handle.net/10541/78134en
dc.description.abstractBACKGROUND: Cervical carcinogenesis is initiated by infection with high-risk (i.e., carcinogenic) human papillomavirus (HPV) types. The subsequent progression from premalignant cervical intraepithelial neoplasia (CIN) to invasive cancer is driven by both genetic and epigenetic processes. We assessed the role of the gene encoding the adhesion molecule tumor suppressor in lung cancer 1 (TSLC1) in this progression. METHODS: We analyzed TSLC1 gene expression by real-time quantitative reverse transcription-polymerase chain reaction, promoter methylation by sodium bisulfite genomic DNA sequencing, and allelic loss by microsatellite analysis in primary keratinocytes, in four non-tumorigenic HPV-immortalized human keratinocyte cell lines, and in 11 human cervical cancer cell lines that were positive for a high-risk HPV DNA type and in normal cervical epithelial cells. We transfected cervical cancer SiHa cells that did not express TSLC1 mRNA with an expression vector containing the TSLC1 complementary DNA (cDNA) or an empty vector and analyzed transfectants for anchorage-independent growth and tumorigenicity in nude mice. We also examined TSLC1 promoter methylation in premalignant cervical lesions and in cervical carcinomas and smears. All statistical tests were two-sided. RESULTS: TSLC1 mRNA was strongly reduced, relative to levels in primary keratinocytes, or absent in 10 (91%) of 11 cervical carcinoma cell lines but in none (0%) of the four HPV-immortalized cell lines (difference = 91%, 95% confidence interval [CI] = 74% to 100%; P =.004). The TSLC1 promoter was hypermethylated, relative to normal foreskin and cervical epithelial cells, in nine (82%) of the 11 cervical carcinoma cell lines but in none (0%) of the four HPV-immortalized cell lines (difference = 82%, 95 CI = 59% to 100%; P =.01). Seven (88%, 95% CI = 47% to 100%) of the eight SiHa/TSLC1 transfectants displayed a marked reduction in anchorage-independent growth (i.e., 0-100 colonies per 5000 cells) compared with none of the four (0%, 95% CI = 0% to 60%) SiHa transfectants bearing the empty vector (i.e., SiHa/hygro transfectants; difference = 88%, 95% CI = 65% to 100%; P =.01) or untransfected SiHa cells. All seven mice (100%, 95% CI = 59% to 100%) injected with untransfected SiHa cells or SiHa/hygro transfectants displayed tumors of at least 50 mm(3) by 2-6 weeks after injection compared with none of eight mice (0%, 95% CI = 0% to 37%) injected with the SiHa/TSLC1 transfectants (difference = 100%, 95% CI = 68% to 100%; P<.001). We detected TSLC1 promoter hypermethylation in seven (35%, 95% CI = 15% to 59%) of 20 high-grade CIN lesions (i.e., CIN II and III) and in 30 (58%, 95% CI = 43% to 71%) of 52 cervical squamous cell carcinomas compared with none (0%, 95% CI = 0% to 34%) of nine normal cervical epithelial biopsy samples and none (0%, 95% CI = 0% to 22%) of 12 CIN I lesions (P<.001 for cervical squamous cell cancer versus normal epithelial biopsy samples plus CIN I lesions). CONCLUSIONS: TSLC1 gene silencing via promoter hypermethylation is a frequent event in the progression from high-risk HPV-containing, high-grade CIN lesions to invasive cervical cancer.en
dc.language.isoenen
dc.subjectUterine Cervical Canceren
dc.subjectCell Line Tumouren
dc.subjectTumour Suppressorsen
dc.subjectTumour Suppressor Proteinsen
dc.subject.meshAnimalsen
dc.subject.meshCell Line, Tumoren
dc.subject.meshCell Transformation, Neoplasticen
dc.subject.meshCervical Intraepithelial Neoplasiaen
dc.subject.meshCervix Uterien
dc.subject.meshChromosomes, Human, Pair 11en
dc.subject.meshCloning, Molecularen
dc.subject.meshDNA Methylationen
dc.subject.meshDNA, Complementaryen
dc.subject.meshDNA, Viralen
dc.subject.meshDown-Regulationen
dc.subject.meshFemaleen
dc.subject.meshGene Expression Regulation, Neoplasticen
dc.subject.meshGene Silencingen
dc.subject.meshGenes, Tumor Suppressoren
dc.subject.meshHumansen
dc.subject.meshImmunoglobulinsen
dc.subject.meshKeratinocytesen
dc.subject.meshLoss of Heterozygosityen
dc.subject.meshMembrane Proteinsen
dc.subject.meshMiceen
dc.subject.meshMice, Nudeen
dc.subject.meshMicrosatellite Repeatsen
dc.subject.meshPapillomaviridaeen
dc.subject.meshPromoter Regions, Geneticen
dc.subject.meshProteinsen
dc.subject.meshRNA, Messengeren
dc.subject.meshReverse Transcriptase Polymerase Chain Reactionen
dc.subject.meshSequence Analysis, DNAen
dc.subject.meshSulfatesen
dc.subject.meshTransfectionen
dc.subject.meshTumor Suppressor Proteinsen
dc.subject.meshUterine Cervical Neoplasmsen
dc.titleTSLC1 gene silencing in cervical cancer cell lines and cervical neoplasia.en
dc.typeArticleen
dc.contributor.departmentDepartment of Pathology, Unit of Molecular Pathology, Vrije Universiteit Medical Center, Amsterdam, The Netherlands. r.steenbergen@vumc.nlen
dc.identifier.journalJournal of the National Cancer Instituteen

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