Evaluation of <i>ZAR1</i> and <i>SFRP4</i> methylation status as potentials biomarkers for diagnosis in cervical cancer: exploratory study phase I

Brebi-Mieville, Priscilla; Hoffstetter, Rene; Andana, Alejandra; Ili, Carmen; Saavedra, Kathleen; Viscarra, Támara; Retamal, Jaime; Sánchez, Raúl; Roa, Juan Carlos

doi:10.3109/1354750x.2013.867535

Cited by 25 publications

(17 citation statements)

References 29 publications

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“…To quantify the methylation level of Reprimo , quantitative methylation-specific PCR (qMSP) was performed. Amplification reactions were carried out in triplicate in a volume of 20 μL that contained 1 μL of bisulfite-modified DNA; 300 nM of each primer; 50 nM probe (Table 1 ); 0.375 U platinum Taq polymerase (Invitrogen); 100 μM of dNTPs; 100 nM ROX dye reference (Invitrogen); 8.3 mM ammonium sulfate; 33.5 mM Trizma (Sigma, St. Louis, MO); 3.35 mM magnesium chloride; 5 mM mercaptoethanol; and 0.05 % DMSO [ 30 ]. Amplifications were carried out using the following profile: 95 °C for 10 min followed by 40 cycles at 95 °C for 30 s, 59 °C for 30 s and 72 °C for 30 s. Amplification reactions were carried out in 96-well plates in the Mx3000P QPCR System (Stratagene).…”

Section: Methodsmentioning

confidence: 99%

Reprimo as a modulator of cell migration and invasion in the MDA-MB-231 breast cancer cell line

et al. 2016

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BackgroundReprimo (RPRM), a highly glycosylated protein, is a new downstream effector of p53-induced cell cycle arrest at the G2/M checkpoint, and a putative tumor suppressor gene frequently silenced via methylation of its promoter region in several malignances. The aim of this study was to characterize the epigenetic inactivation and its biological function in BC cell lines.MethodsThe correlation between RPRM methylation and loss of mRNA expression was assessed in six breast cancer cell lines by methylation specific PCR (MSP), 5′-Aza-2′-deoxycytidine treatment and RT-PCR assays. MDA-MB-231 cells were chosen to investigate the phenotypic effect of RPRM in cell proliferation, cell cycle, cell death, cell migration and invasion.ResultsIn the cancer methylome system (CMS) (web-based system for visualizing and analyzing genome-wide methylation data of human cancers), the CpG island region of RPRM (1.1 kb) was hypermethylated in breast cancer compared to normal breast tissue; more interesting still was that ERα(+) tumors showed higher methylation intensity than ERα(−). Downregulation of RPRM mRNA by methylation was confirmed in MDA-MB-231 and BT-20 cell lines. In addition, overexpression of RPRM in MDA-MB-231 cells resulted in decreased rates of cell migration, wound healing and invasion in vitro. However, RPRM overexpression did not alter cell viability, phosphatidylserine (PS) translocation or G2/M cell cycle transition.ConclusionTaken together, these data suggest that RPRM is involved in decreased cell migration and invasion in vitro, acting as a potential tumor suppressor gene in the MDA-MB-231 cell line.

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Section: Methodsmentioning

confidence: 99%

Reprimo as a modulator of cell migration and invasion in the MDA-MB-231 breast cancer cell line

et al. 2016

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“…In normal adult cells, it binds to Wnt morphogens and prevents their binding to corresponding transmembrane Frizzled receptor, thereby impeding the canonical Wnt pathway. However, loss of expression of sFRP4 due to promoter hypermethylation has been found in several types of cancer, such as colorectal cancer [6], bladder cancer [7], mesothelioma [8], cervical cancer [9], and ovarian cancer [10]. This promotes the upregulation of Wnt signaling, resulting in stabilization of a cytoplasmic pool of b-catenin protein, which is the signaling node of the canonical Wnt pathway.…”

Section: Introductionmentioning

confidence: 99%

Antagonizing canonical Wnt signaling pathway by recombinant human sFRP4 purified from E. coli and its implications in cancer therapy

Ghoshal

Ghosh

2016

Mol Cell Biochem

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The Wnt signaling pathway plays a predominant role in aberrant proliferation in myriad of cancers. In non-cancerous cells, Wnts are blocked by the secreted frizzled-related proteins (sFRPs) that are generally downregulated in cancer cells. We have purified and characterized bacterially expressed glutathione S-transferase-tagged SFRP4 from a novel clone generated from human cell origin. Cervical cancer (HeLa) and lung cancer (A549) cells, in which Wnt and associated genes were found to be expressed, were treated with the purified recombinant sFRP4, which revealed a significant dose-dependent cell growth inhibition up to 40 %. The current investigation on functionality of this bacterially produced recombinant sFRP4 in arresting cancer cell proliferation is the first of its kind, where G2/M phase arrest and early apoptosis were evident. Increase in phosphorylated β-catenin in sFRP4 treatment indicated inhibition of Wnt pathway, which was further confirmed by downregulation of pro-proliferative genes, namely cyclin D1, c-myc, and survivin. Functional activity of recombinant sFRP4 was further exploited in co-therapy module with chemotherapeutic drugs to decipher molecular events. Collectively, our study on purified recombinant sFRP4 from bacterial host holds great promise in targeting Wnt signaling for exploring new strategies to combat cancer.

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“…Hypermethylation of the SFRP4 promoter was associated with CC and may have utility for the molecular screening of cervical neoplasia (38). Brebi et al (39) revealed that SFRP4 may be used as a potential biomarker for CC diagnosis. Therefore, SFRP4 was identified to be closely associated with CC.…”

Section: Discussionmentioning

confidence: 99%

Systematic prediction of target genes and pathways in cervical cancer from microRNA expression data

et al. 2018

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Abstract. Cervical cancer (CC) is a leading cause of cancer-associated mortality in women; thus, the present study aimed to investigated potential target genes and pathways in patients with CC by utilizing an ensemble method and pathway enrichment analysis. The ensemble method integrated a correlation method [Pearson's correlation coefficient (PCC)], a causal inference method (IDA) and a regression method [least absolute shrinkage and selection operator (Lasso)] using the Borda count election algorithm, forming the PCC, IDA and Lasso (PIL) method. Subsequently, the PIL method was validated to be a feasible approach to predict microRNA (miRNA) targets by comparing predicted miRNA targets against those from a confirmed database. Finally, Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis was conducted for target genes in the 1,000 most frequently predicted miRNA-mRNA interactions to determine target pathways. A total of 10 target genes were obtained that were predicted >5 times, including secreted frizzled-related protein 4, maternally expressed 3 and NIPA like domain containing 4. Additionally, a total of 17 target pathways were identified, of which cytokine-cytokine receptor interaction (P=8.91x10 -7 ) was the most significantly associated with CC of all pathways. In conclusion, the present study predicted target genes and pathways for patients with CC based on miRNA expression data, the PIL method and pathway analysis. The results of the present study may provide an insight into the pathological mechanisms underlying CC, and provide potential biomarkers for the diagnosis and treatment of this tumor type. However, these biomarkers have yet to be validated; these validations will be performed in future studies.

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Evaluation of ZAR1 and SFRP4 methylation status as potentials biomarkers for diagnosis in cervical cancer: exploratory study phase I

Abstract: ZAR1 and SFRP4 qMSP could be used as potential biomarker for CC diagnosis.

Cited by 25 publications

References 29 publications

Reprimo as a modulator of cell migration and invasion in the MDA-MB-231 breast cancer cell line

Reprimo as a modulator of cell migration and invasion in the MDA-MB-231 breast cancer cell line

Antagonizing canonical Wnt signaling pathway by recombinant human sFRP4 purified from E. coli and its implications in cancer therapy

Systematic prediction of target genes and pathways in cervical cancer from microRNA expression data

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