Gene Expression Analysis of Preinvasive and Invasive Cervical Squamous Cell Carcinomas Identifies <i>HOXC10</i> as a Key Mediator of Invasion

Zhai, Yali; Kuick, Rork; Nan, Bin; Ota, Ichiro; Weiss, Stephen J.; Trimble, Cornelia L.; Fearon, Eric R.; Cho, Kathleen R.

doi:10.1158/0008-5472.can-07-2056

Cited by 222 publications

(241 citation statements)

References 47 publications

(47 reference statements)

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“…HPRT is a commonly used reference gene, previously used for the study of low-level cytokine expression [17] and cervical cancer [40]. HPRT has been verified as an optimal normalizer between samples in colon, prostate, breast, skin, and bladder tissue [41], but had not been established as a suitable normalizer between normal and diseased cervical tissue.…”

Section: Discussionmentioning

confidence: 99%

Gene expression analysis of interferon κ in laser capture microdissected cervical epithelium

DeCarlo

Escott

Werner

et al. 2008

Analytical Biochemistry

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Optimal sample handling techniques for tissue preparation and storage, RNA extraction and quantification, and target gene detection are crucial for reliable gene expression analysis. Methods for measuring low-expressing genes, such as interferons, in human cervical samples are not described in the scientific literature. To detect interferon mRNA in human cervical samples we obtained normal and dysplastic frozen and formalin-fixed cervical biopsies from colposcopy. Histopathological diagnosis was performed by one pathologist. Cervical keratinocytes were isolated using laser capture microdissection. Immortalized keratinocytes transduced with or devoid of an HPV oncogene were used for initial method development. RNA from samples was extracted and integrity tested to compare tissue storage and extraction methods. The expression of five housekeeping genes was analyzed in cell lines and patient tissue to permit normalization between samples using quantitative real-time polymerase chain reaction. The usefulness of cDNA amplification was assessed for the detection of low-expressing interferon κ in cervical tissue. Here we report optimal tissue storage conditions, RNA extraction, sample normalization, and transcript amplification, as well as the sensitivity of quantitative real-time polymerase chain reaction and laser capture microdissection, for interferon κ detection in cervical tissue. Without these optimized techniques, interferon κ detection would be unattainable in cervical samples. KeywordsCervical keratinocytes; Frozen and formalin-fixed cervical tissue; Housekeeping genes; Interferons; Interferon κ; Laser capture microdissection; Quantitative real-time polymerase chain reaction Quantitative real-time polymerase chain reaction (qRT-PCR) 1 for the molecular analysis of disease is a powerful and widely used tool. Extraction of high-quality RNA for use in gene expression analysis techniques such as reverse transcription (RT), qRT-PCR, and cDNA microarrays is of great importance. Although obtaining high-quality RNA from cell lines is relatively straightforward, the complexity and heterogeneity of human tissue pose considerable challenges for linking gene expression patterns with disease state. Nonetheless,

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Section: Discussionmentioning

confidence: 99%

Gene expression analysis of interferon κ in laser capture microdissected cervical epithelium

DeCarlo

Escott

Werner

et al. 2008

Analytical Biochemistry

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“…Indeed, several homeobox genes (HOX) have been identified as bona fide oncogenes. For example, HOXC10 overexpression is associated with increased invasion and the transition of cervical high-grade squamous intraepithelial lesions to cervical squamous cell carcinoma (28). HOXB7 mediates aspects of epithelial-to-mesenchymal transition in breast cancer, promotes proliferation in oral cancer, and is associated with progression and development of metastases in lung cancer (29)(30)(31).…”

Section: Cervical Cancer Cells Are Sensitive To Treatment With the Kdmentioning

confidence: 99%

Tumor suppressor p16 ^INK4A is necessary for survival of cervical carcinoma cell lines

McLaughlin-Drubin

Park²,

Münger

2013

Proc. Natl. Acad. Sci. U.S.A.

153

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The tumor suppressor p16 INK4A inhibits formation of enzymatically active complexes of cyclin-dependent kinases 4 and 6 (CDK4/6) with D-type cyclins. Oncogenic stress induces p16 INK4A expression, which in turn triggers cellular senescence through activation of the retinoblastoma tumor suppressor. Subversion of oncogene-induced senescence is a key step during cancer development, and many tumors have lost p16 INK4A activity by mutation or epigenetic silencing. Human papillomavirus (HPV)-associated tumors express high levels of p16 INK4A in response to E7 oncoprotein expression. Induction of p16 INK4A expression is not a consequence of retinoblastoma tumor suppressor inactivation but is triggered by a cellular senescence response and is mediated by epigenetic derepression through the H3K27-specific demethylase (KDM)6B. HPV E7 expression causes an acute dependence on KDM6B expression for cell survival. The p16 INK4A tumor suppressor is a critical KDM6B downstream transcriptional target and its expression is critical for cell survival. This oncogenic p16 INK4A activity depends on inhibition of CDK4/CDK6, suggesting that in cervical cancer cells where retinoblastoma tumor suppressor is inactivated, CDK4/CDK6 activity needs to be inhibited in order for cells to survive. Finally, we note that HPV E7 expression creates a unique cellular vulnerability to small-molecule KDM6A/B inhibitors.synthetic lethality | apoptosis | biomarker | cancer therapy P osttranslational histone modifications, including phosphorylation, ubiquitination, acetylation, and methylation, impact both the physical state and the transcriptional competence of chromatin. These modifications are dynamic and regulate a variety of cellular processes, such as stem cell maintenance, cell fate determination and maintenance, cell cycle control, and epigenetic heritability of transcriptional programs (reviewed in refs. 1 and 2). Methylation of lysine residues on histones is modulated by histone lysine methyltransferases (KMTs) and histone lysine demethylases (KDMs). Although histone lysine methylation is involved in both transcriptional activation and repression, histone H3 trimethylation of lysine 27 (H3K27me3) on gene promoters is crucial for epigenetic silencing mediated by polycomb group proteins (3-5). The JmjC-domain containing histone demethylases, KDM6A (UTX) and KDM6B (JMJD3) remove this repressive mark, allowing for transcriptional activation (6-10). Although KDM6A and KDM6B appear identical with regards to catalytic activities and histone substrate specificities, they have a number of nonoverlapping and nonredundant biological activities. KDM6B-but not KDM6A-regulates RAS/RAF-mediated oncogene-induced senescence (OIS) (11, 12). OIS was originally described as a cell-abortive response to ectopic RAS oncogene expression in normal human cells. It is now recognized that OIS represents one of several cell-intrinsic tumor-suppressor responses that function to eliminate aberrantly proliferating, potentially premalignant cells. Evidence for OIS has been dete...

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“…some tumors, aberrant Hox gene expressions directly drives tumorigenesis through escape from apoptosis [10], alterations to receptor signalling [11], tumor cell invasion [12], and epithelial-mesenchymal transition (EMT) [13].…”

mentioning

confidence: 99%

Upregulated Hoxc6 expression is associated with poor survival in gastric cancer patients

Zhang¹,

Jin²,

Yang³

et al. 2013

neo

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Human Hox genes (Homeobox) have crucial roles in development and differentiation, regulating numerous processes including apoptosis, receptor signalling, differentiation, motility and angiogenesis. Aberrant expression of Hoxc6 gene has been reported in several tumor tissues and cancer cell lines. The prognostic significance of Hoxc6 in gastric cancer remains largely unknown.This study was aimed to investigate the clinical significance of Hoxc6 in gastric cancer.Total RNA of paired tissue samples (n=25) and a tissue microarray containing 161 paired tissues from patients with gastric cancers at different stages were collected. Quantitative real-time PCR and immunochemistry assay were carried out to investigate the expression of Hoxc6.Hoxc6 mRNA was increased in gastric cancer tissues ( 16 of 25) compared with the adjacent normal mucosa (P<0.05). Immunohistochemical detection showed that expression of Hoxc6 was associated with the depth of tumor invasion (P<0.05). Patients with higher expression levels of Hoxc6 had a shorter overall survival rate (P<0.05).Hoxc6 might contribute to the progression of gastric carcinogenesis and may be a significant predictor of poor survival in patients with gastric cancer after curative operations.

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Gene Expression Analysis of Preinvasive and Invasive Cervical Squamous Cell Carcinomas Identifies HOXC10 as a Key Mediator of Invasion

Cited by 222 publications

References 47 publications

Gene expression analysis of interferon κ in laser capture microdissected cervical epithelium

Gene expression analysis of interferon κ in laser capture microdissected cervical epithelium

Tumor suppressor p16 ^INK4A is necessary for survival of cervical carcinoma cell lines

Upregulated Hoxc6 expression is associated with poor survival in gastric cancer patients

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Gene Expression Analysis of Preinvasive and Invasive Cervical Squamous Cell Carcinomas Identifies HOXC10 as a Key Mediator of Invasion

Cited by 222 publications

References 47 publications

Gene expression analysis of interferon κ in laser capture microdissected cervical epithelium

Gene expression analysis of interferon κ in laser capture microdissected cervical epithelium

Tumor suppressor p16 INK4A is necessary for survival of cervical carcinoma cell lines

Upregulated Hoxc6 expression is associated with poor survival in gastric cancer patients

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Tumor suppressor p16 ^INK4A is necessary for survival of cervical carcinoma cell lines