Ds. Byun et al., Frequent epigenetic inactivation of RASSF1A by aberrant promoter hypermethylation in human gastric adenocarcinoma, CANCER RES, 61(19), 2001, pp. 7034-7038
Methylation associated inactivation of RASSF1, a putative tumor suppressor
identified at 3p21.3, has been frequently observed in several human maligna
ncies, including lung and breast cancers. To explore the penetrance of RASS
F1 in gastric carcinogenesis, we performed expression and mutation analyses
of 3 isotypes; of RASSF1 (A, B, and C) in 150 gastric specimens, including
15 carcinoma cell lines. RASSF1A and RASSF1B transcripts were not expresse
d in 60% (9 of 15) and 33% (5 of 15) of gastric carcinoma cell lines, respe
ctively, whereas RASSF1C was detectable in all cell fines. Bisulfite DNA se
quencing analysis revealed that the CpG island in the RASSF1A promoter is h
ypermethylated in all RASSF1A-nonexpressing cell lines. In addition, both R
ASSF1A and RASSF1B were re-expressed by treatment with the demethylating ag
ent 5-aza-2'-deoxycytidine. Among 90 primary gastric adenocarcinomas examin
ed, 41 (46%) and 19 (21%) expressed no or abnormally low levels of RASSF1A
and RASSF1B, respectively, and 12 (13%) tumors showed no expression of both
isoforms. Loss or abnormal down-regulation of RASSF1A correlated with tumo
r stage and grade but not with histological types of tumors. Methylation-sp
ecific PCR analysis demonstrated that 95% (39 of 41) of RASSF1A-nonexpressi
ng primary tumors are methylated at the CpG sites in the promoter, whereas
none of the adjacent noncancerous or normal tissues are methylated. No soma
tic mutations were detected in RASSF1 transcripts expressed in unmethylated
tumors. However, 10 methylated tumors, including 4 cell lines, showed low
genomic levels of RASSF1 and expressed no RASSF1A transcripts, suggesting t
hat RASSF1A inactivation might be caused by both epigenetic and genetic mec
hanisms in a subset of gastric adenocarcinomas. In conclusion, our data ind
icate that epigenetic transcriptional silencing of RASSF1, especially RASSF
1A isoform, is a frequent event in gastric tumorigenesis and might play an
important role in the malignant progression of gastric adenocarcinomas.