CAJ | 학술논문

目的：探讨 HPV16E6/E7对 KYSE450细胞中凋亡调控因子 Bcl-2和 Bad 表达的影响。方法采用瞬时转染技术将 HPV1E6/E7融合基因的真核表达载体转染食管癌细胞株 KYSE450中，分组用 TSA（HDACi）处理后，收集 RNA，应用反转录-聚合酶链反应（reverse transcription polymerase chain reaction RT-PCR）技术检测Bcl-2、Bad 的 mRNA 表达情况。结果（1）用 RT-PCR 检测转染后的结果为阳性。（2）Bad mRNA 在食管癌细胞KYSE450中表达较低，E6E7过表达对细胞内 Bad 基因 mRNA 水平无影响，各转染组之间没有差异。（3）Bcl-2 mRNA 在食管癌细胞 KYSE450中均有表达，E6/E7对 Bcl-2有轻度调控作用。（4）HDAC 抑制剂 TSA 处理后， TSA 明显增加了 E6对 Bcl-2基因表达的诱导调控作用。结论KYSE450细胞中 E6和 E7基因促进 Bcl-2在转录水平上的表达，而对 Bad 的表达无明显的影响。在病毒感染鳞状上皮细胞后早期表达致癌基因 E6、E7可能诱导凋亡调控因子 Bcl-2在转录水平上的表达。
목적：탐토 HPV16E6/E7대 KYSE450세포중조망조공인자 Bcl-2화 Bad 표체적영향。방법채용순시전염기술장 HPV1E6/E7융합기인적진핵표체재체전염식관암세포주 KYSE450중，분조용 TSA（HDACi）처리후，수집 RNA，응용반전록-취합매련반응（reverse transcription polymerase chain reaction RT-PCR）기술검측Bcl-2、Bad 적 mRNA 표체정황。결과（1）용 RT-PCR 검측전염후적결과위양성。（2）Bad mRNA 재식관암세포KYSE450중표체교저，E6E7과표체대세포내 Bad 기인 mRNA 수평무영향，각전염조지간몰유차이。（3）Bcl-2 mRNA 재식관암세포 KYSE450중균유표체，E6/E7대 Bcl-2유경도조공작용。（4）HDAC 억제제 TSA 처리후， TSA 명현증가료 E6대 Bcl-2기인표체적유도조공작용。결론KYSE450세포중 E6화 E7기인촉진 Bcl-2재전록수평상적표체，이대 Bad 적표체무명현적영향。재병독감염린상상피세포후조기표체치암기인 E6、E7가능유도조망조공인자 Bcl-2재전록수평상적표체。
Objective To investigate the human papillomavirus 16 (HPV16 )E6E7′s impact on the expres-sion of apoptosis regulation genes Bcl-2 and Bad in esophageal squamous cancer cell line KYSE-450.Meth-ods E6E7 fusion eukaryotic expression vector was transfected into the esophageal cell lines KYSE450 by transient transfection technique,and grouped with TSA(HDACi)treatment;the expressions of Bcl-2 and Bad were detected by reverse transcription polymerase chain reaction RT-PCR.Results (1)The result af-ter transfection was positive by RT-PCR.(2)There was low expression of Bad gene in KYSE-450 cell lines;E6E7overexpression had no effect on Bad mRNA and there was no any difference between the groups.(3)Bcl-2 gene was expressed in KYSE-450 cell lines,suggesting that Bcl-2 was slightly regulated by E6E7.(4)Treatment after TSA(HDACi),which significantly increase the regulation of E6 on the Bcl-2 expres-sion.Conclusion E6,E7 genes in KYSE450 cell promote Bcl-2 expression at the transcriptional level ,while they have no any effect on the expression of Bad .Therefore,the viral infection of early squamous cells oncogenes E6, E7 may induce the expression of apoptosis regulation gene Bcl-2 at the transcriptional level.