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Identification of differentially-expressed genes by DNA methylation in cervical cancer.

Lee HS, Yun JH, Jung J, Yang Y, Kim BJ, Lee SJ, Yoon JH, Moon Y, Kim JM, Kwon YI - Oncol Lett (2015)

Bottom Line: A number of the identified genes were novel cervical cancer-related genes and their differential expression was confirmed in a publicly available database.From these results, the expression of the CAMK2N1, ALDH1A3 and PPP1R3C genes are were shown to be suppressed in cervical cancers by DNA methylation.These genes may be involved in the progression or initiation of cervical cancer.

View Article: PubMed Central - PubMed

Affiliation: Center for Genome Science, Korea National Institute of Health, Osong Health Technology Administration Complex, Cheongju, Chungcheongbuk-do 363-951, Republic of Korea.

ABSTRACT

To identify novel cervical cancer-related genes that are regulated by DNA methylation, integrated analyses of genome-wide DNA methylation and RNA expression profiles were performed using the normal and tumor regions of tissues from four patients; two with cervical cancer and two with pre-invasive cancer. The present study identified 19 novel cervical cancer-related genes showing differential RNA expression by DNA methylation. A number of the identified genes were novel cervical cancer-related genes and their differential expression was confirmed in a publicly available database. Among the candidate genes, the epigenetic regulation and expression of three genes, CAMK2N1, ALDH1A3 and PPP1R3C, was validated in HeLa cells treated with a demethylating reagent using methylation-specific polymerase chain reaction (PCR) and quantitative PCR, respectively. From these results, the expression of the CAMK2N1, ALDH1A3 and PPP1R3C genes are were shown to be suppressed in cervical cancers by DNA methylation. These genes may be involved in the progression or initiation of cervical cancer.

No MeSH data available.


Related in: MedlinePlus

Validation of epigenetic regulation of selected genes in cervical cancer cells. HeLa cells were plated and treated with various concentrations of 5-aza-2′-deoxycytidine as decitabine for 96 h. Purified genomic DNA was modified by converting cytosine residues to uracil, and methylation-specific-PCR analysis was conducted with methylation- and unmethylation-specific primers for each gene. Reverse transcription was performed with total RNA, and quantitative PCR analysis was performed. PCR, polymerase chain reaction.
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f2-ol-09-04-1691: Validation of epigenetic regulation of selected genes in cervical cancer cells. HeLa cells were plated and treated with various concentrations of 5-aza-2′-deoxycytidine as decitabine for 96 h. Purified genomic DNA was modified by converting cytosine residues to uracil, and methylation-specific-PCR analysis was conducted with methylation- and unmethylation-specific primers for each gene. Reverse transcription was performed with total RNA, and quantitative PCR analysis was performed. PCR, polymerase chain reaction.

Mentions: To further confirm whether the selected genes were regulated by DNA methylation, the HeLa cells were treated with a demethylating reagent, and the methylation status and RNA expression level was measured by MS-PCR using each primer set (Table III) and RT-quantitative PCR analysis, respectively. Methylation of selected genes, CAMK2N1, ALDH1A3 and PPP1R3C, was found to be decreased, while demethylation was increased following treatment with the demethylating reagent, as expected. Additionally, RNA expression was increased dose-dependently as a result of the decreased methylation status (Fig. 2). These results indicated that the genes were originally methylated in cervical cancer cells and showed increased demethylation followed by increased RNA expression following treatment with a demethylating reagent.


Identification of differentially-expressed genes by DNA methylation in cervical cancer.

Lee HS, Yun JH, Jung J, Yang Y, Kim BJ, Lee SJ, Yoon JH, Moon Y, Kim JM, Kwon YI - Oncol Lett (2015)

Validation of epigenetic regulation of selected genes in cervical cancer cells. HeLa cells were plated and treated with various concentrations of 5-aza-2′-deoxycytidine as decitabine for 96 h. Purified genomic DNA was modified by converting cytosine residues to uracil, and methylation-specific-PCR analysis was conducted with methylation- and unmethylation-specific primers for each gene. Reverse transcription was performed with total RNA, and quantitative PCR analysis was performed. PCR, polymerase chain reaction.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4356325&req=5

f2-ol-09-04-1691: Validation of epigenetic regulation of selected genes in cervical cancer cells. HeLa cells were plated and treated with various concentrations of 5-aza-2′-deoxycytidine as decitabine for 96 h. Purified genomic DNA was modified by converting cytosine residues to uracil, and methylation-specific-PCR analysis was conducted with methylation- and unmethylation-specific primers for each gene. Reverse transcription was performed with total RNA, and quantitative PCR analysis was performed. PCR, polymerase chain reaction.
Mentions: To further confirm whether the selected genes were regulated by DNA methylation, the HeLa cells were treated with a demethylating reagent, and the methylation status and RNA expression level was measured by MS-PCR using each primer set (Table III) and RT-quantitative PCR analysis, respectively. Methylation of selected genes, CAMK2N1, ALDH1A3 and PPP1R3C, was found to be decreased, while demethylation was increased following treatment with the demethylating reagent, as expected. Additionally, RNA expression was increased dose-dependently as a result of the decreased methylation status (Fig. 2). These results indicated that the genes were originally methylated in cervical cancer cells and showed increased demethylation followed by increased RNA expression following treatment with a demethylating reagent.

Bottom Line: A number of the identified genes were novel cervical cancer-related genes and their differential expression was confirmed in a publicly available database.From these results, the expression of the CAMK2N1, ALDH1A3 and PPP1R3C genes are were shown to be suppressed in cervical cancers by DNA methylation.These genes may be involved in the progression or initiation of cervical cancer.

View Article: PubMed Central - PubMed

Affiliation: Center for Genome Science, Korea National Institute of Health, Osong Health Technology Administration Complex, Cheongju, Chungcheongbuk-do 363-951, Republic of Korea.

ABSTRACT

To identify novel cervical cancer-related genes that are regulated by DNA methylation, integrated analyses of genome-wide DNA methylation and RNA expression profiles were performed using the normal and tumor regions of tissues from four patients; two with cervical cancer and two with pre-invasive cancer. The present study identified 19 novel cervical cancer-related genes showing differential RNA expression by DNA methylation. A number of the identified genes were novel cervical cancer-related genes and their differential expression was confirmed in a publicly available database. Among the candidate genes, the epigenetic regulation and expression of three genes, CAMK2N1, ALDH1A3 and PPP1R3C, was validated in HeLa cells treated with a demethylating reagent using methylation-specific polymerase chain reaction (PCR) and quantitative PCR, respectively. From these results, the expression of the CAMK2N1, ALDH1A3 and PPP1R3C genes are were shown to be suppressed in cervical cancers by DNA methylation. These genes may be involved in the progression or initiation of cervical cancer.

No MeSH data available.


Related in: MedlinePlus