J Korean Med Sci.  2007 Sep;22(Suppl):S32-S37. 10.3346/jkms.2007.22.S.S32.

Hypermethylation of p16(INK4a) in Korean Non-small Cell Lung Cancer Patients

• Hong YS
• Roh MS
• Kim NY
• Lee HJ
• Kim HK
• Lee KE
• Kwak JY
• Kim JY

• KMID: 1785786
• DOI: http://doi.org/10.3346/jkms.2007.22.S.S32

Abstract

Promoter hypermethylation of the p16(INK4a) gene was investigated in 81 sets of samples of tumor tissue and adjacent normal tissue from Korean patients with primary lung cancer, using the modified real-time polymerase chain reaction (PCR)/ SYBR Green detection method. The results showed hypermethylation of p16(INK4a) in 27.2% of tumor tissues, and in 11.1% of adjacent normal tissue. No significant association was found between the overall aberrant methylation in tumor and corresponding normal specimens (r=0.137, p=0.219). In 22 cases with p16(INK4a) hypermethylation in tumor tissues, only 4 (18.1%) cases were found to have a hypermethylated normal tissue specimen. The findings of this study show that smoking can influence the methylation level of the promoter region of p16(INK4a), and that this occurs in tumor tissues more frequently than in normal tissues. Other clinicopathological characteristics, including age, sex, tumor stage, and histologic type were not found to be correlated with p16(INK4a) methylation.

Keyword

Hypermethylation; p16(INK4a); Lung Cancer

MeSH Terms

Adult
Aged
Base Sequence
Carcinoma, Non-Small-Cell Lung/*genetics
*DNA Methylation
DNA Primers/genetics
DNA, Neoplasm/chemistry/genetics
Female
*Genes, p16
Humans
Korea
Lung Neoplasms/*genetics
Male
Middle Aged
Molecular Sequence Data
Polymerase Chain Reaction
Promoter Regions, Genetic
Smoking/adverse effects/genetics

Figure

• 

  Fig. 1 Methylation analysis of the p16INK4a promoter sequencing (Genebank accession number X94154). The positions of the CCGG sites are underlined. Each primer is shown in bold characters.

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  Fig. 2 Results of p16INK4a gene promoter methylation of normal (A) and tumor (B) lung tissues by real-time PCR. 2, 3, 5: Normal lung tissues (A); 2*, 3*, 5*: tumor lung tissues (B); 1, 4, 6: positive control (wi-38); 7: negative control (water); a: no-cut DNA amplification; b: Hpa II-cut DNA amplification; c: Msp I-cut DNA amplification. Delta Rn: the magnitude of the fluorescence signal generated during the PCR at each time point.

• 

  Fig. 3 Result of serial dilutions to determine the detection limits of the real-time PCR protocol showing the initial DNA amounts used in the amplification.

• 

  Fig. 4 Methylation status of the p16INK4a gene prompter in normal and tumor tissues. The black circle denotes methylation positivity, and the open circle indicates that the sample was negative for methylation. N, normal tissue; T, tumor tissue.

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