Background:EBV DNA is found within the malignant cells of 10% of gastric cancers. Modern molecular technology facilitates identification of virus-related biochemical effects that could assist in early diagnosis and disease management.
Methods:In this study, RNA expression profiling was performed on 326 macrodissected paraffin-embedded tissues including 204 cancers and, when available, adjacent non-malignant mucosa. Nanostring nCounter probes targeted 96 RNAs (20 viral, 73 human, and 3 spiked RNAs).
Results:In 182 tissues with adequate housekeeper RNAs, distinct profiles were found in infectedversusuninfected cancers, and in malignantversusadjacent benign mucosa. EBV-infected gastric cancers expressed nearly all of the 18 latent and lytic EBV RNAs in the test panel. Levels ofEBER1andEBER2RNA were highest and were proportional to the quantity of EBV genomes as measured by Q-PCR. Among protein coding EBV RNAs,EBNA1from the Q promoter andBRLF1were highly expressed whileEBNA2levels were low positive in only 6/14 infected cancers. Concomitant upregulation of cellular factors implies that virus is not an innocent bystander but rather is linked to NFKB signaling (FCER2, TRAF1) and immune response(TNFSF9, CXCL11, IFITM1, FCRL3, MS4A1 and PLUNC), withPPARGexpression implicating altered cellular metabolism. Compared to adjacent non-malignant mucosa, gastric cancers consistently expressedINHBA, SPP1, THY1, SERPINH1, CXCL1, FSCN1, PTGS2 (COX2), BBC3, ICAM1, TNFSF9, SULF1, SLC2A1, TYMS, three collagens, the cell proliferation markersMYCandPCNA, and EBVBLLF1while they lackedCDH1 (E-cadherin), CLDN18,PTEN, SDC1(CD138),GAST(gastrin) and its downstream effectorCHGA(chromogranin). Compared to lymphoepithelioma-like carcinoma of the uterine cervix, gastric cancers expressedCLDN18, EPCAM, REG4, BBC3, OLFM4, PPARG, andCDH17while they had diminished levels ofIFITM1andHIF1A. The druggable targets ERBB2 (Her2), MET, and the HIF pathway, as well as several other potential pharmacogenetic indicators (including EBV infection itself, as well asSPARC, TYMS, FCGR2BandREG4) were identified in some tumor specimens.
Conclusion:This study shows how modern molecular technology applied to archival fixed tissues yields novel insights into viral oncogenesis that could be useful in managing affected patients.