Aberrant promoter hypermethylation and silencing of the critical 3p21 tumour suppressor gene, RASSF1A, in Chinese oesophageal squamous cell carcinoma

3p21 is an important locus harbouring critical tumour suppressor genes (TSG), which are implicated in the pathogenesis of multiple tumours, including oesophageal carcinoma. RASSF1A is a 3p21.3 candidate TSG frequently inactivated by promoter methylation in multiple tumours. We investigated RASSF1A p...

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Veröffentlicht in:International journal of oncology 2006-03, Vol.28 (3), p.767-773
Hauptverfasser: LY WONG, Michelle, QIAN TAO, TSAO, George S. W, LAM, King-Yin, LAW, Simon, WONG, John, SRIVASTAVA, Gopesh, LI FU, WONG, Kai-Yau, QIU, Guo-Hua, LAW, Fian B. F, TIN, Pui-Chi, CHEUNG, Wai-Ling, LEE, Ping-Yin, TANG, Johnny Cheuk-On
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container_issue 3
container_start_page 767
container_title International journal of oncology
container_volume 28
creator LY WONG, Michelle
QIAN TAO
TSAO, George S. W
LAM, King-Yin
LAW, Simon
WONG, John
SRIVASTAVA, Gopesh
LI FU
WONG, Kai-Yau
QIU, Guo-Hua
LAW, Fian B. F
TIN, Pui-Chi
CHEUNG, Wai-Ling
LEE, Ping-Yin
TANG, Johnny Cheuk-On
description 3p21 is an important locus harbouring critical tumour suppressor genes (TSG), which are implicated in the pathogenesis of multiple tumours, including oesophageal carcinoma. RASSF1A is a 3p21.3 candidate TSG frequently inactivated by promoter methylation in multiple tumours. We investigated RASSF1A promoter methylation and gene expression in Chinese oesophageal squamous cell carcinoma (ESCC) to compare it to data from Japanese patients. Methylation-specific PCR (MSP) showed that RASSF1A was partially methylated in 3/7 (43%) cell lines; 22/64 (34%) primary tumours and 3/64 (5%) corresponding non-tumour samples; and was not methylated in 2 immortalized normal oesophageal epithelial cell lines and 6 normal oesophageal epithelium samples. Bisulfite genome sequencing confirmed the MSP results. Promoter hypermethylation correlated well with RASSF1A mRNA down-regulation. Treatment of cell lines with 5-aza-2'-deoxycytidine activated RASSF1A mRNA expression along with promoter demethylation. RASSF1A hypermethylation in the Chinese cohort was much lower than in a published report of Japanese ESCC patients (52%) and cell lines (74%). Our own analysis of Japanese ESCC cell lines for direct comparison also detected a high frequency of RASSF1A hypermethylation (8/10; 80%) and high levels of hypermethylation at each CpG site. No significant association between RASSF1A hypermethylation and histological differentiation (p=0.953), tumour staging (p=0.117), or survival (p=0.7571) was found in Chinese ESCC, unlike the results of Japanese patients. The incidence of oesophageal cancer shows marked variation by geographic area and ethnic group; it is almost three times higher in China than in Japan, indicating possible different pathogenetic mechanisms. Our results show that RASSF1A hypermethylation in ESCC has epidemiological/ethnic differences, and suggest that Chinese ESCC may result from different pathogenetic mechanisms.
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W ; LAM, King-Yin ; LAW, Simon ; WONG, John ; SRIVASTAVA, Gopesh ; LI FU ; WONG, Kai-Yau ; QIU, Guo-Hua ; LAW, Fian B. F ; TIN, Pui-Chi ; CHEUNG, Wai-Ling ; LEE, Ping-Yin ; TANG, Johnny Cheuk-On</creator><creatorcontrib>LY WONG, Michelle ; QIAN TAO ; TSAO, George S. W ; LAM, King-Yin ; LAW, Simon ; WONG, John ; SRIVASTAVA, Gopesh ; LI FU ; WONG, Kai-Yau ; QIU, Guo-Hua ; LAW, Fian B. F ; TIN, Pui-Chi ; CHEUNG, Wai-Ling ; LEE, Ping-Yin ; TANG, Johnny Cheuk-On</creatorcontrib><description>3p21 is an important locus harbouring critical tumour suppressor genes (TSG), which are implicated in the pathogenesis of multiple tumours, including oesophageal carcinoma. RASSF1A is a 3p21.3 candidate TSG frequently inactivated by promoter methylation in multiple tumours. We investigated RASSF1A promoter methylation and gene expression in Chinese oesophageal squamous cell carcinoma (ESCC) to compare it to data from Japanese patients. Methylation-specific PCR (MSP) showed that RASSF1A was partially methylated in 3/7 (43%) cell lines; 22/64 (34%) primary tumours and 3/64 (5%) corresponding non-tumour samples; and was not methylated in 2 immortalized normal oesophageal epithelial cell lines and 6 normal oesophageal epithelium samples. Bisulfite genome sequencing confirmed the MSP results. Promoter hypermethylation correlated well with RASSF1A mRNA down-regulation. Treatment of cell lines with 5-aza-2'-deoxycytidine activated RASSF1A mRNA expression along with promoter demethylation. RASSF1A hypermethylation in the Chinese cohort was much lower than in a published report of Japanese ESCC patients (52%) and cell lines (74%). Our own analysis of Japanese ESCC cell lines for direct comparison also detected a high frequency of RASSF1A hypermethylation (8/10; 80%) and high levels of hypermethylation at each CpG site. No significant association between RASSF1A hypermethylation and histological differentiation (p=0.953), tumour staging (p=0.117), or survival (p=0.7571) was found in Chinese ESCC, unlike the results of Japanese patients. The incidence of oesophageal cancer shows marked variation by geographic area and ethnic group; it is almost three times higher in China than in Japan, indicating possible different pathogenetic mechanisms. 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Methylation-specific PCR (MSP) showed that RASSF1A was partially methylated in 3/7 (43%) cell lines; 22/64 (34%) primary tumours and 3/64 (5%) corresponding non-tumour samples; and was not methylated in 2 immortalized normal oesophageal epithelial cell lines and 6 normal oesophageal epithelium samples. Bisulfite genome sequencing confirmed the MSP results. Promoter hypermethylation correlated well with RASSF1A mRNA down-regulation. Treatment of cell lines with 5-aza-2'-deoxycytidine activated RASSF1A mRNA expression along with promoter demethylation. RASSF1A hypermethylation in the Chinese cohort was much lower than in a published report of Japanese ESCC patients (52%) and cell lines (74%). Our own analysis of Japanese ESCC cell lines for direct comparison also detected a high frequency of RASSF1A hypermethylation (8/10; 80%) and high levels of hypermethylation at each CpG site. 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RASSF1A is a 3p21.3 candidate TSG frequently inactivated by promoter methylation in multiple tumours. We investigated RASSF1A promoter methylation and gene expression in Chinese oesophageal squamous cell carcinoma (ESCC) to compare it to data from Japanese patients. Methylation-specific PCR (MSP) showed that RASSF1A was partially methylated in 3/7 (43%) cell lines; 22/64 (34%) primary tumours and 3/64 (5%) corresponding non-tumour samples; and was not methylated in 2 immortalized normal oesophageal epithelial cell lines and 6 normal oesophageal epithelium samples. Bisulfite genome sequencing confirmed the MSP results. Promoter hypermethylation correlated well with RASSF1A mRNA down-regulation. Treatment of cell lines with 5-aza-2'-deoxycytidine activated RASSF1A mRNA expression along with promoter demethylation. RASSF1A hypermethylation in the Chinese cohort was much lower than in a published report of Japanese ESCC patients (52%) and cell lines (74%). Our own analysis of Japanese ESCC cell lines for direct comparison also detected a high frequency of RASSF1A hypermethylation (8/10; 80%) and high levels of hypermethylation at each CpG site. No significant association between RASSF1A hypermethylation and histological differentiation (p=0.953), tumour staging (p=0.117), or survival (p=0.7571) was found in Chinese ESCC, unlike the results of Japanese patients. The incidence of oesophageal cancer shows marked variation by geographic area and ethnic group; it is almost three times higher in China than in Japan, indicating possible different pathogenetic mechanisms. Our results show that RASSF1A hypermethylation in ESCC has epidemiological/ethnic differences, and suggest that Chinese ESCC may result from different pathogenetic mechanisms.</abstract><cop>Athens</cop><pub>Editorial Academy of the International Journal of Oncology</pub><pmid>16465383</pmid><doi>10.3892/ijo.28.3.767</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record>
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source Spandidos Publications Journals; MEDLINE; EZB-FREE-00999 freely available EZB journals
subjects Biological and medical sciences
Carcinoma, Squamous Cell - epidemiology
Carcinoma, Squamous Cell - genetics
Carcinoma, Squamous Cell - pathology
Cell Line, Tumor
China - epidemiology
Chromosomes, Human, Pair 3 - genetics
DNA Methylation
Esophageal Neoplasms - epidemiology
Esophageal Neoplasms - genetics
Esophageal Neoplasms - pathology
Esophagus
Gastroenterology. Liver. Pancreas. Abdomen
Genes, Tumor Suppressor
Humans
Incidence
Japan - epidemiology
Medical sciences
Neoplasm Staging
Promoter Regions, Genetic - genetics
Survival Analysis
Tumor Suppressor Proteins - genetics
Tumors
title Aberrant promoter hypermethylation and silencing of the critical 3p21 tumour suppressor gene, RASSF1A, in Chinese oesophageal squamous cell carcinoma
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