Adenovirus-mediated transfer of HPV 16 E6/E7 antisense RNA to human cervical cancer cells.
Adenovirus-mediated transfer of HPV 16 E6/E7 antisense RNA to human cervical cancer cells.
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腺病毒介导的 HPV 16 E6/E7 反义 RNA 转移至人宫颈癌细胞。
DOI：
10.1006/gyno.1996.0310
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Mitchell,MF
中科院分区：
文献类型：
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作者：
Hamada,K;Sakaue,M;Alemany,R;Zhang,WW;Horio,Y;Roth,JA;Mitchell,MF
为探讨腺病毒反义RNA转录物在宫颈癌基因治疗中的应用前景，我们将人乳头瘤病毒（HPV）16型E6和E7基因的反义RNA转录物通过重组腺病毒载体Ad 5CMV-HPV 16 AS导入携带HPV 16型的宫颈癌细胞中，分析了这些基因的表达对细胞生长和肿瘤生长的影响。Ad 5CMV-HPV 16 AS含有巨细胞病毒启动子、HPV 16的E6和E7基因的反义序列和SV 40多聚腺苷酸化信号，并将其插入到腺病毒5的E1缺失区。用聚合酶链反应（PCR）扩增HPV 16 E6/E7区全长，克隆入微基因盒。逆转录PCR检测到HPV 16 E6/E7反义RNA在Ad 5CMV-HPV 16 AS感染的SiHa细胞中的表达。腺病毒p53能显著增强Ad 5CMV-HPV 16 AS对SiHa细胞的生长抑制作用。这些数据表明，HPV 16 E6/E7反义RNA以Ad 5CMV-HPV 16 AS的形式转染宫颈癌细胞是一种潜在的治疗HPV 16阳性宫颈癌的新方法。
To explore the potential of an adenoviral antisense RNA transcript for gene therapy of cervical cancer, we introduced the antisense RNA transcript ofE6andE7genes of human papillomavirus (HPV) 16 into cervical cancer cells harboring HPV 16 via a recombinant adenoviral vector, Ad5CMV–HPV 16 ASand analyzed the effects of expression of these genes on cell growth and tumor growth. Ad5CMV–HPV 16 AScontains the cytomegalovirus-promoter,E6andE7genes of HPV 16 in antisense orientation, and the SV40 polyadenylation signal in a mini-gene cassette, which is inserted into theE1-deleted region of modified adenovirus 5. The entireE6/E7region of HPV 16 was amplified by polymerase chain reaction (PCR) before cloning into the mini-gene cassette. By reverse transcriptase–PCR, HPV 16E6/E7antisense RNA was detected in SiHa cells infected with Ad5CMV–HPV 16 AS.The growth of the Ad5CMV–HPV 16 AS-infected cells was greatly suppressed, as evidenced by a decrease in cell count. The growth inhibitory effect of Ad5CMV–HPV 16 ASwas significantly enhanced by an adenoviralp53construct, Ad5CMV–p53.In anex vivostudy in nude mice, tumorigenicity was completely inhibited in mice injected with Ad5CMV–HPV 16 AS-infected SiHa cells. These data suggest that transfection of cervical cancer cells with HPV 16E6/E7antisense RNA in a form such as Ad5CMV–HPV 16 ASis a potential novel approach to the therapy of HPV 16-positive cervical cancer.