The Study of Integrated EBV DNA in the Genome of NPC Cell Line H2B17-7

• Main Authors: Yi-Chen Kuo, 郭宜蓁
• Other Authors: Sou-De Cheng
• Format: Others
• Language: zh-TW
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/09764444424195987651

碩士 === 長庚大學 === 基礎醫學研究所 === 95 === EBV has been known to be correlated with nasopharyngeal carcinoma (NPC). The used cell line H2B17-7 was modified from NPC cell line HONE-1 followed by transfecting with EBV particles. This study aims to investigate the nature of integrated EBV DNA fragment, and the integration sites in human genome. The cell line was found harbouring no episomal EBV DNA, and proving that several EBV DNA fragments are integrated in one of its chromosomes by fluorescence in situ hybridization. With Southern analyses, the genome of H2B17-7 shows positive results after examined with multiple EBV probes, including BamHI C, W, O, Z, R, K, B, G, D, b, V, and A fragments. Other than the regular size bands, an irregular fragment was also hybridized by BamHI C and W probes. To examine this particular fragment, a partial genomic library from BamHI fragments 4~6 kb was made with ZAP Express® vector. After further screening this library with W probe, a specific clone was selected. After consequent DNA sequencing, this 4589 bp DNA insert contains part of BamHI Q and part of W fragments connecting with each other and directing reversely. The BamHI Y, H, and F regions between the two are eliminated. The region from terminal repeat to BamHI C, and part region of BamHI W are also not included in this insert. Further investigating of this integrated sequence in the genome of H2B17-7 shall lead to the flanking human sequence, which may consequently reveals the EBV integration sites at human genome, and the possible impact to the host cells.