Correlation of Epstein-Barr virus and its encoded proteins with Helicobacter pylori and expression of c-met and c-myc in gastric carcinoma.
OBJECTIVE
To investigate the interrelationship of Epstein-Barr virus (EBV) and EBV- encoded proteins with Helicobacter pylori (H pylori) infection and the expression of c-met and c-myc oncogene proteins in gastric carcinoma, and to explore their role in gastric carcinogenesis.


METHODS
One hundred and eighty-five gastric carcinoma tissues were detected by polymerase chain reaction (PCR)-Southern blot for EBV genome and in situ hybridization (ISH) for EBV-encoded small RNA 1 (EBER1). Gastric carcinoma with positive EBER1 signals was confirmed EBV-associated gastric carcinoma (EBVaGC). The status of H pylori infection in 185 gastric carcinomas was assessed by rapid urease test and PCR. The samples with positive PCR and urease test were defined as H pylori infection. The expression of c-met and c-myc oncogene proteins in tissues of EBVaGC and matched EBV-negative gastric carcinoma (EBVnGC) were examined by immunohistochemistry. RT-PCR and Southern hybridization were used to detect the expression of nuclear antigens (EBNAs) 1 and 2, latent membrane protein (LMP) 1, early genes BARF1 and BHRF1 in EBVaGC cases.


RESULTS
The positive rate of H pylori and EBV in 185 gastric carcinomas was 59.45% (110/185) and 7.03% (13/185) respectively. No difference was found in sex, age, pathological differentiation, clinical stages and lymph node metastasis between H pylori-positive and H pylori-negative gastric carcinomas. However, the positive rate of H pylori infection in the antrum gastric carcinomas was higher than that of cardia and body gastric carcinomas. In our series, age, pathological differentiation, clinical stages, lymph node metastasis and location of cancer were not different between EBVnGC and EBVaGC, while the positive rate of EBV in male patients was significantly higher than that of female patients. The positivity of H pylori in EBV-associated and EBV-negative gastric carcinomas was 46.15% (6/13) and 81.40%(104/172) respectively. There was no significant correlation between EBV and H pylori infection. The c-met overexpression was significantly higher in the EBVaGC group than in the EBVnGC group. However, c-met and c-myc expression did not show significant difference between the two groups. Transcripts of EBNA1 were detected in all 13 EBVaGCs, while both EBNA2 and LMP1 mRNA were not detected. Six of the 13 cases exhibited BARF1 transcripts and 2 exhibited BHRF1 transcripts.


CONCLUSIONS
The positivity of H pylori in EBVnGCs is higher than that of EBVaGCs, but no significant correlation is found between EBV infection and H pylori infection. H pylori-positive gastric carcinoma is predominant in antrum location, while EBVaGC has a tendency of predominance in cardia/body location. EBV infection is associated with c-met abnormal expression but not with c-myc protein in EBVaGC. c-met overexpression is not induced by LMP1. BARF1 and BHRF1 may play important roles in the tumorigenesis of EBVaGC through different pathways.
Luo
Bing
B
Department of Microbiology, Qingdao University Medical College, 38 Dengzhou Road, Qingdao 266021, Shandong Province, China. qdluobing@yahoo.com
Wang
Yun
Y
Wang
Xiao-Feng
XF
Gao
Yu
Y
Huang
Bao-Hua
BH
Zhao
Peng
P
D000328:Adult; D000368:Aged; D001426:Bacterial Proteins / Q000032:analysis; D020031:Epstein-Barr Virus Infections / Q000503:physiopathology*; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics; D005260:Female; D015964:Gene Expression Regulation, Bacterial; D015972:Gene Expression Regulation, Neoplastic; D015967:Gene Expression Regulation, Viral; D016481:Helicobacter Infections / Q000503:physiopathology*; D016480:Helicobacter pylori / Q000302:isolation & purification*; D004854:Herpesvirus 4, Human / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008875:Middle Aged; D019859:Proto-Oncogene Proteins c-met / Q000235:genetics*; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics; D013270:Stomach / Q000382:microbiology / Q000473:pathology / Q000821:virology; D013274:Stomach Neoplasms / Q000235:genetics* / Q000382:microbiology* / Q000821:virology; D014764:Viral Proteins / Q000032:analysis / Q000235:genetics
C086495:BARF1 protein, Human herpesvirus 4; C083278:BHRF1 protein, Human herpesvirus 4; D001426:Bacterial Proteins; D019309:Epstein-Barr Virus Nuclear Antigens; C099537:Epstein-Barr virus encoded RNA 1; D016271:Proto-Oncogene Proteins c-myc; D012367:RNA, Viral; D014764:Viral Proteins; D019859:Proto-Oncogene Proteins c-met; C417029:EBV-encoded nuclear antigen 1