Differential expression of Epstein Barr virus-encoded miRNAs in nasopharyngeal carcinoma cell line C666-1 and CNE-2Z

Objective:Aberrant expression of microRNAs(miRNAs) is involved in the progression of carcinomas.Epstein Barr virus(EBV) encodes near 40 miRNAs.This study was aimed to investigate the differential expression of EBV-encoded miRNAs in two poor-differentiated nasopharyngeal carcinomas(NPC) cell line C666-1 and CNE-2Z,and to analyze targets of the ebv-miRNAs.Methods:Human NPC cell line C666-1(EBV+) and CNE-2Z(EBV may lose due to the mass passages) were cultured,total RNA was isolated and examined.mRNAs of LMP-1 and LMP-2A were assessed by RT-PCR to identify the infection of EBV,and the expression profiles of ebv-miRNAs was detected with miRNA microarray chip and compared array results was confirmed by real-time PCR.Ebv-miRNA targets were analyzed by literature review.Results:The purification of total RNA from C666-1 and CNE-2Z were high,and the expression of mRNAs of LMP-1 and LMP-2A was detected in C666-1 and CNE-2Z.According to microarray screening,19/39 ebv-miRNAs were differentially expressed.Real-time PCR revealed that ebv-miR-BHRF1-1 and ebv-miR-BART14* were up-regulated in C666-1,while ebv-miR-BART8* was down-regulated,these results were coincidence with the tendency of microarray.Up to date,limitable ebv-miRNA targets involved in cell signaling,transcription,proliferation,immunoreactions,apoptosis and viral DNA reproduction were confirmed or predicted by informatics analysis.Conclusion:Ebv-miRNAs may be involved in the pathogenesis of NPC by regulating their target genes.Differential expression of ebv-miRNAs in NPC cells implies the possibility of individual treatment of NPC targeting on EBV miRNAs.