Methylation of p16INK4a is a non-rare event in cervical intraepithelial neoplasia

Suki Kang, Jungsik Kim, Hong Bae Kim, Jung Won Shim, Eunji Nam, Sung Hoon Kim, Hee Jung Ahn, Yoon Pyo Choi, Boxiao Ding, Kijun Song, Namhoon Cho

Research output: Contribution to journalArticle

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Abstract

The cell cycle inhibitor, p16INK4a may be a useful surrogate biomarker of cervical intraepithelial neoplasia (CIN); however, there is currently no consensus of p16INK4a genetic alterations throughout the multiple step process of CIN. Our goal was to identify the methylation frequency of p16INK4a in each step of CIN that is associated with human papillomavirus (HPV) infection, using several different detection methods of p16INK4a methylation to correlate the data. The present study included a total of 43 patients, including 38 with CIN, and 5 normal patients. Three different methods were used to detect hypermethylation of CpG islands, methylation-specific PCR (MSP) amplification of different primer sets of M1, M2, and M3, pyrosequencing of each forward primer region, and immunohistochemistry of p16INK4a. Analysis of MSP showed that 20 of the 38 CIN patients (52.6%) revealed hypermethylation in at least 1 primer set of the p16 INK4a promoter. A complete loss of p16INK4a protein expression was observed in 11 cases (28.9%). There was no observed association of methylation of the p16INK4a gene with either CIN grading (P=0.0698) or HPV status (P=0.2811): specifically 42.9% (3/7) was found in CIN 1, 57.1% (8/14) in CIN 2, and 52.9% (9/17) in CIN 3. In concordance with immunohistochemistry results, hypermethylation of the p16INK4a promoter was significantly correlated with a lack of p16 protein expression (P=0.0145). All positive peaks from pyrosequencing matched the MSP results, which ranged from 6.3% to 24.5%. In conclusion, p16INK4a gene silencing during CIN was not determined to be a particularly rare event; however, it does not correlate with either HPV status or CIN grading.

Original languageEnglish
Pages (from-to)74-82
Number of pages9
JournalDiagnostic Molecular Pathology
Volume15
Issue number2
DOIs
Publication statusPublished - 2006 Jun 1

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Cervical Intraepithelial Neoplasia
Methylation
p16 Genes
Polymerase Chain Reaction
Immunohistochemistry
Cyclin-Dependent Kinase Inhibitor p16
CpG Islands
Papillomavirus Infections
Gene Silencing
Cell Cycle
Biomarkers

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Molecular Biology
  • Cell Biology

Cite this

Kang, Suki ; Kim, Jungsik ; Kim, Hong Bae ; Shim, Jung Won ; Nam, Eunji ; Kim, Sung Hoon ; Ahn, Hee Jung ; Choi, Yoon Pyo ; Ding, Boxiao ; Song, Kijun ; Cho, Namhoon. / Methylation of p16INK4a is a non-rare event in cervical intraepithelial neoplasia. In: Diagnostic Molecular Pathology. 2006 ; Vol. 15, No. 2. pp. 74-82.
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title = "Methylation of p16INK4a is a non-rare event in cervical intraepithelial neoplasia",
abstract = "The cell cycle inhibitor, p16INK4a may be a useful surrogate biomarker of cervical intraepithelial neoplasia (CIN); however, there is currently no consensus of p16INK4a genetic alterations throughout the multiple step process of CIN. Our goal was to identify the methylation frequency of p16INK4a in each step of CIN that is associated with human papillomavirus (HPV) infection, using several different detection methods of p16INK4a methylation to correlate the data. The present study included a total of 43 patients, including 38 with CIN, and 5 normal patients. Three different methods were used to detect hypermethylation of CpG islands, methylation-specific PCR (MSP) amplification of different primer sets of M1, M2, and M3, pyrosequencing of each forward primer region, and immunohistochemistry of p16INK4a. Analysis of MSP showed that 20 of the 38 CIN patients (52.6{\%}) revealed hypermethylation in at least 1 primer set of the p16 INK4a promoter. A complete loss of p16INK4a protein expression was observed in 11 cases (28.9{\%}). There was no observed association of methylation of the p16INK4a gene with either CIN grading (P=0.0698) or HPV status (P=0.2811): specifically 42.9{\%} (3/7) was found in CIN 1, 57.1{\%} (8/14) in CIN 2, and 52.9{\%} (9/17) in CIN 3. In concordance with immunohistochemistry results, hypermethylation of the p16INK4a promoter was significantly correlated with a lack of p16 protein expression (P=0.0145). All positive peaks from pyrosequencing matched the MSP results, which ranged from 6.3{\%} to 24.5{\%}. In conclusion, p16INK4a gene silencing during CIN was not determined to be a particularly rare event; however, it does not correlate with either HPV status or CIN grading.",
author = "Suki Kang and Jungsik Kim and Kim, {Hong Bae} and Shim, {Jung Won} and Eunji Nam and Kim, {Sung Hoon} and Ahn, {Hee Jung} and Choi, {Yoon Pyo} and Boxiao Ding and Kijun Song and Namhoon Cho",
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Kang, S, Kim, J, Kim, HB, Shim, JW, Nam, E, Kim, SH, Ahn, HJ, Choi, YP, Ding, B, Song, K & Cho, N 2006, 'Methylation of p16INK4a is a non-rare event in cervical intraepithelial neoplasia', Diagnostic Molecular Pathology, vol. 15, no. 2, pp. 74-82. https://doi.org/10.1097/00019606-200606000-00003

Methylation of p16INK4a is a non-rare event in cervical intraepithelial neoplasia. / Kang, Suki; Kim, Jungsik; Kim, Hong Bae; Shim, Jung Won; Nam, Eunji; Kim, Sung Hoon; Ahn, Hee Jung; Choi, Yoon Pyo; Ding, Boxiao; Song, Kijun; Cho, Namhoon.

In: Diagnostic Molecular Pathology, Vol. 15, No. 2, 01.06.2006, p. 74-82.

Research output: Contribution to journalArticle

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T1 - Methylation of p16INK4a is a non-rare event in cervical intraepithelial neoplasia

AU - Kang, Suki

AU - Kim, Jungsik

AU - Kim, Hong Bae

AU - Shim, Jung Won

AU - Nam, Eunji

AU - Kim, Sung Hoon

AU - Ahn, Hee Jung

AU - Choi, Yoon Pyo

AU - Ding, Boxiao

AU - Song, Kijun

AU - Cho, Namhoon

PY - 2006/6/1

Y1 - 2006/6/1

N2 - The cell cycle inhibitor, p16INK4a may be a useful surrogate biomarker of cervical intraepithelial neoplasia (CIN); however, there is currently no consensus of p16INK4a genetic alterations throughout the multiple step process of CIN. Our goal was to identify the methylation frequency of p16INK4a in each step of CIN that is associated with human papillomavirus (HPV) infection, using several different detection methods of p16INK4a methylation to correlate the data. The present study included a total of 43 patients, including 38 with CIN, and 5 normal patients. Three different methods were used to detect hypermethylation of CpG islands, methylation-specific PCR (MSP) amplification of different primer sets of M1, M2, and M3, pyrosequencing of each forward primer region, and immunohistochemistry of p16INK4a. Analysis of MSP showed that 20 of the 38 CIN patients (52.6%) revealed hypermethylation in at least 1 primer set of the p16 INK4a promoter. A complete loss of p16INK4a protein expression was observed in 11 cases (28.9%). There was no observed association of methylation of the p16INK4a gene with either CIN grading (P=0.0698) or HPV status (P=0.2811): specifically 42.9% (3/7) was found in CIN 1, 57.1% (8/14) in CIN 2, and 52.9% (9/17) in CIN 3. In concordance with immunohistochemistry results, hypermethylation of the p16INK4a promoter was significantly correlated with a lack of p16 protein expression (P=0.0145). All positive peaks from pyrosequencing matched the MSP results, which ranged from 6.3% to 24.5%. In conclusion, p16INK4a gene silencing during CIN was not determined to be a particularly rare event; however, it does not correlate with either HPV status or CIN grading.

AB - The cell cycle inhibitor, p16INK4a may be a useful surrogate biomarker of cervical intraepithelial neoplasia (CIN); however, there is currently no consensus of p16INK4a genetic alterations throughout the multiple step process of CIN. Our goal was to identify the methylation frequency of p16INK4a in each step of CIN that is associated with human papillomavirus (HPV) infection, using several different detection methods of p16INK4a methylation to correlate the data. The present study included a total of 43 patients, including 38 with CIN, and 5 normal patients. Three different methods were used to detect hypermethylation of CpG islands, methylation-specific PCR (MSP) amplification of different primer sets of M1, M2, and M3, pyrosequencing of each forward primer region, and immunohistochemistry of p16INK4a. Analysis of MSP showed that 20 of the 38 CIN patients (52.6%) revealed hypermethylation in at least 1 primer set of the p16 INK4a promoter. A complete loss of p16INK4a protein expression was observed in 11 cases (28.9%). There was no observed association of methylation of the p16INK4a gene with either CIN grading (P=0.0698) or HPV status (P=0.2811): specifically 42.9% (3/7) was found in CIN 1, 57.1% (8/14) in CIN 2, and 52.9% (9/17) in CIN 3. In concordance with immunohistochemistry results, hypermethylation of the p16INK4a promoter was significantly correlated with a lack of p16 protein expression (P=0.0145). All positive peaks from pyrosequencing matched the MSP results, which ranged from 6.3% to 24.5%. In conclusion, p16INK4a gene silencing during CIN was not determined to be a particularly rare event; however, it does not correlate with either HPV status or CIN grading.

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