Analytical Cytology Laboratory and the Key Immunopathology Laboratory of Guangdong Province, Shantou University Medical College, Shantou, People's Republic of China.
Biochem Cell Biol. 2010 Aug;88(4):697-704. doi: 10.1139/O10-005.
The objective of this investigation was to determine if simultaneous silencing of the human papillomavirus type 18 (HPV-18) E6 and E7 oncogenes using RNA interference (RNAi) would be a potential therapeutic approach against the carcinogenic activity of this virus. Two synthetic double-stranded oligonucleotides, encoding short hairpin transcripts corresponding to HPV-18 E6 and E7 genes, were cloned into pGenesilence (pGS) 1.0 vectors to produce pGS-E6, pGS-E7, and pGS-(E6+E7), respectively. Our results showed that the expression of HPV-18 E6 class 1 and HPV-18 E7 in HeLa cells was markedly decreased after being transfected with pGS-E6, pGS-E7, and pGS-(E6+E7) vectors. Of the three vectors, pGS-(E6+E7) had a greater ability to decrease the growth rate of HeLa cells, inhibit colony formation in soft agar, and significantly reduce tumor growth in nude mice. We also found that depletion of HPV-18 E6 and E7 in this manner promoted apoptosis of HeLa cells. Our data showed that simultaneously decreasing HPV-18 E6 and E7 gene expression in HeLa cells by RNAi could significantly inhibit tumor growth under in vitro conditions and in nude mice. These data suggest that gene therapy may be a possible therapeutic approach for HPV-positive cervical cancers.
本研究旨在探讨应用 RNA 干扰(RNAi)同时沉默人乳头瘤病毒 18 型(HPV-18)E6 和 E7 癌基因是否可能成为针对该病毒致癌活性的一种潜在治疗方法。我们将分别编码 HPV-18 E6 和 E7 基因短发夹转录本的两条合成双链寡核苷酸克隆到 pGenesilence(pGS)1.0 载体中,以构建 pGS-E6、pGS-E7 和 pGS-(E6+E7)载体。结果表明,转染 pGS-E6、pGS-E7 和 pGS-(E6+E7)载体后,HeLa 细胞中 HPV-18 E6 类 1 和 HPV-18 E7 的表达明显下降。在这三种载体中,pGS-(E6+E7)对 HeLa 细胞生长速度的抑制、软琼脂集落形成的抑制和裸鼠肿瘤生长的抑制作用最强。我们还发现,以这种方式耗尽 HPV-18 E6 和 E7 可促进 HeLa 细胞凋亡。我们的数据表明,应用 RNAi 同时降低 HeLa 细胞中 HPV-18 E6 和 E7 基因的表达可显著抑制体外和裸鼠条件下的肿瘤生长。这些数据提示基因治疗可能成为 HPV 阳性宫颈癌的一种潜在治疗方法。