EB病毒对人胚鼻咽上皮细胞的转化

In vitro transformation of human embryonic nasopharyngeal epithelial cells with Epstein-Barr virus

为观察EBV和／或促癌物四癸酸佛波醇二酯（TPA）对其的转化作用，以人胚鼻咽上皮作体外原代组织培养，采用自B95－8细胞分离的EB病毒直接感染或结合TPA处理体外培养的人胚鼻咽上皮细胞，着重观察感染细胞在半固体培养基中的集落形成率；并采用PCR扩增法探讨EB病毒是否直接进入鼻咽上皮细胞。结果显示：单独EB病毒或灭活（56℃，30分钟）EB病毒加TPA感染时，病毒不能进入细胞导致表型改变；活性EB病毒结合TPA同时处理或先用EB病毒后用TPA处理时，EB病毒能直接进入细胞并导致细胞集落形成率明显增高（P＜0．05）。从而表明EB病毒体外能部分转化人胚鼻咽上皮细胞，其转化作用依赖于TPA的存在和病毒基因组的完整。

EBV from B95-8 cells were used to infect directly the human embryonic nasopharyngealepithelial (HENE) cells in vitro. Results: Primary HENE cells treated with EBV did not have a signifi cantly increased colony-forming rate in soft agarose, while cells that were treated with EBV in combina tion with tumor promotor 12 - 0-tetradecanoylphorbol - 13 acetat e (TPA ) sh owed a m a rk ed inc rease incolony forming rate from 0-5 cells to 20-4O colonies per 10000 cells. (2) HENE cells treated with inacti vated (56℃ for 30 minutes )virus, in spite of adding TPA at the same time, did n0t significantly in crease agar0se colony forming rate; (3) HENE cells treated with TPA after EBV exposure for a weekhad significantly increased colony forming rate in soft aragose, while the cells exposed to EBV after us ing TPA treatment did not. In addition, EBV BNLF1 (LMP) fragment in HENE cells treated withEBV or EBV+TPA was detected by polymerase chain reaction using specific oligonucleotide primer.Only HENE cells treated with EBV+TPA presented positive band in aragose gel elecrophoresis. Theresults suggest that transformation of EBV on HENE cells depends on infectious virus and an intact viralgen0me. TPA not only can promote the transformation, but also promote EBV's entering HENE cells.