Inactivation mechanisms and growth suppressive effects of p16INK4a in Asian esophageal squamous carcinoma cell lines.
The inactivation mechanisms and functional role of p16INK4a in three Asian esophageal squamous cell carcinoma (ESCC) cell lines were investigated by polymerase chain reaction (PCR) amplification, DNA sequencing, methylation-specific PCR analysis, reverse transcription-PCR, Western blotting, and colony formation assays. The p16INK4a was inactivated by promoter hypermethylation in all three cell lines, a homozygous deletion of exons 2 and 3, and a frameshift deletion on exon 1, leading to transcriptional silencing or the production of mutant p16INK4a protein. Two ESCC cell lines transfected with wild type p16INK4a show significantly reduced cell growth properties. The results of the present studies support the suppressive role of p16INK4a in ESCC development.
Kwong
Fung Mei
FM
Department of Biology, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong (SAR), People's Republic of China.
Tang
Johnny Cheuk On
JC
Srivastava
Gopesh
G
Lung
Maria Li
ML
D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell / Q000235:genetics* / Q000517:prevention & control; D045744:Cell Line, Tumor; D019175:DNA Methylation; D004938:Esophageal Neoplasms / Q000235:genetics* / Q000517:prevention & control; D019942:Genes, p16 / Q000502:physiology*; D006801:Humans; D019656:Loss of Heterozygosity; D016133:Polymerase Chain Reaction