Promoter methylation of HIN-1 in the progression to esophageal squamous cancer.
HIN-1 (High in normal-1) is a tumor suppressor gene that is highly expressed in many epithelial tissues, including breast lung, trachea, pancreas, prostrate and salivary gland. Inactivation of HIN-1 expression by promoter methylation is frequent in many epithelial carcinomas and carcinoma-in-situ, including breast, lung and nasopharyngeal. Because HIN-1 silencing commences at an early stage of malignant transformation in these tissues, it may be a useful marker for tumor presence. The status of HIN-1 regulation in esophageal cancer has not been previously reported. Thus, we analyzed 18 esophageal cancer cell lines for HIN-1 expression and methylation by reverse transcription PCR (RT-PCR) and methylation specific PCR (MSP). HIN-1 gene silencing and promoter methylation was present in 13 (72%) of the cell lines. Bisulfite-treated sequencing confirmed the methylation status in cell lines and demonstrated dense methylation of HIN-1 throughout the promoter region. Epigenetic changes of HIN-1 were examined throughout the progression of carcinogenesis in esophageal squamous lesions through analysis of archived surgical specimens from patients with normal esophageal mucosa (n = 17), grade I dysplasia (n = 39), grade II dysplasia (n = 12), grade III dysplasia (n = 9), and invasive squamous esophageal cancer (n = 126). Methylation of HIN-1 was present in 0% of normal mucosa, 31% of grade I dysplasia, 33% of grade II dysplasia, 44% of grade III dysplasia, and 50% of esophageal cancer specimens analyzed. These studies demonstrate HIN-1 silencing is associated with dense promoter region hypermethylation in esophageal cancer and suggest that methylation of HIN-1 is an early event in dysplastic transformation.
Guo
Mingzhou
M
Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, Maryland, USA.
Ren
Jingli
J
Brock
Malcolm V
MV
Herman
James G
JG
Carraway
Hetty E
HE
D000483:Alleles; D045744:Cell Line, Tumor; D016207:Cytokines / Q000235:genetics* / Q000378:metabolism; D019175:DNA Methylation; D018450:Disease Progression; D004938:Esophageal Neoplasms / Q000235:genetics* / Q000473:pathology*; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D006801:Humans; D018307:Neoplasms, Squamous Cell / Q000235:genetics* / Q000473:pathology*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics*; D017422:Sequence Analysis, DNA; D025521:Tumor Suppressor Proteins / Q000235:genetics* / Q000378:metabolism
D016207:Cytokines; C434732:SCGB3A1 protein, human; D025521:Tumor Suppressor Proteins