The role of human papillomavirus 16 E6 in anchorage-independent and invasive growth of mouse tonsil epithelium
The role of human papillomavirus 16 E6 in anchorage-independent and invasive growth of mouse tonsil epithelium
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DOI：
10.1001/archotol.133.5.495
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发表时间：
2007-05-01
影响因子：
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通讯作者：
Lee, John H.
中科院分区：
文献类型：
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作者：
Hoover, Andrew C.;Spanos, William C.;Lee, John H.
目的：提供一个可操作的系统来研究人乳头瘤病毒 16 (HPV16) E6 相关的人类受 HPV16 影响的上皮细胞类型的转化机制。设计：对用 HPV16 癌基因加 H-ras 转化的小鼠扁桃体上皮细胞 (MTEC) 进行体外和体内生化和生理学研究。设置：基本 研究实验室。参与者：C57BL/6 小鼠。干预措施：将逆转录病毒载体中的 HPV16 癌基因 E6 和 E7 转导至 MTEC，并分离表达 E6、E7 或单独表达 E6、E7 或两者的单独克隆或与 H-ras 联合表达的单个克隆。主要结果指标：培养物中的生长、贴壁独立生长以及免疫活性、同基因中的生长 结果：表达 E6 的 MTEC 通过蛋白酶体阻断抑制的机制降解 p53。尽管正常MTEC在20次群体倍增后衰老，但单独使用E6或与E7组合足以使MTEC永生化超过25次群体倍增，缩短其群体倍增时间，并允许不依赖贴壁的生长。然而，只有表达 E6 加 H-ras 或 E6/E7 加 H-ras 的 MTEC 在免疫活性、同基因小鼠的原位口腔内和皮下部位形成侵袭性肿瘤。结论：我们发现单独的 HPV16 E6 和 E7 不足以进行侵袭性生长。然而，H-ras 和 E6 的协同活性足以导致侵袭性生长。
Objective: To provide a manipulatable system to study the mechanism of human papillomavirus 16 (HPV16) E6-related transformation of an epithelial cell type affected by HPV16 in humans.Design: Biochemical and physiological studies of mouse tonsil epithelial cells (MTECs) transformed with HPV16 oncogenes plus H-ras in vitro and in vivo.Setting: Basic research laboratory.Participants: C57BL/6 mice.Interventions: Transduction of the HPV16 oncogenes E6 and E7 in retroviral vectors into MTECs with isolation of multiple individual clones that expressed E6, E7, or both alone or in conjunction with H-ras.Main Outcome Measures: Growth in culture, anchorage-independent growth, and growth in immune competent, syngeneic mice.Results: The MTECs that expressed E6 degraded p53 by a mechanism that is inhibited by proteasomal blockade. Although normal MTECs senesced after 20 population doublings, E6 alone or in combination with E7 was sufficient to immortalize MTECs beyond 25 population doublings, lower their population-doubling time, and permit anchorage-independent growth. However, only MTECs that express E6 plus H-ras or E6/E7 plus H-ras formed invasive tumors in immune competent, syngeneic mice at orthotopic intraoral and subdermal sites.Conclusions: We found that HPV16 E6 and E7 alone are not sufficient for invasive growth. However, the synergistic activity of H-ras and E6 was sufficient to result in invasive growth.