Al Moustafa Ala-Eddin, Foulkes William D, Wong Annick, Jallal Houda, Batist Gerald, Yu Qunyan, Herlyn Meenhard, Sicinski Piotr, Alaoui-Jamali Moulay A
Program in Cancer Genetics, McGill University, Montreal, Quebec, Canada.
Oncogene. 2004 Jul 1;23(30):5252-6. doi: 10.1038/sj.onc.1207679.
More than 25% of head and neck squamous cell carcinomas (HNSCC) and 99% of cervical cancers (CxCa) are positive for high-risk human papillomaviruses (HPVs). Furthermore, the type I tyrosine kinase receptor ErbB-2 is overexpressed in at least 30% of HNSCC and CxCa. Recently, we demonstrated that E6/E7 of HPV type 16 cooperate with ErbB-2 to induce cell transformation of human normal oral epithelial (NOE) cells. This is accompanied by overexpression of cyclin D1 in NOE cells. To determine the role of cyclin D1 in E6/E7/ErbB-2 cooperation, we examined the independent effects of E6/E7 and ErbB-2, and the combined effect of E6/E7 and ErbB-2 in mouse normal embryonic fibroblast (NEF), wild type (wt), and knockout cyclin D1 (D1(-/-)) cells. We report that NEF-wt cells transduced with E6/E7 alone and E6/E7/ErbB-2 together form small and large tumors in nude mice, respectively, as well as different sized colonies in soft agar; whereas ErbB-2 alone elicits neither tumor formation in vivo nor colony formation in soft agar. More importantly, E6/E7, ErbB-2 and E6/E7/ErbB-2 together all fail to induce neoplastic transformation of cyclin D1(-/-) cells in vivo and in vitro. Furthermore, using antisense cyclin D1 we completely inhibited tumor and colony formation of NEF-wt-E6/E7 and wt-E6/E7-ErbB-2 as well as human NOE-E6/E7-ErbB-2-transformed cells. These analyses reveal that cyclin D1 is the downstream target of the neoplastic transformation induced by E6/E7 or E6/E7/ErbB-2 cooperation in normal cells. Our data suggest that anti-cyclin D1 therapy may be highly specific in the treatment of all human cancers expressing high-risk HPVs or HPVs/ErbB-2.
超过25%的头颈部鳞状细胞癌（HNSCC）和99%的宫颈癌（CxCa）高危型人乳头瘤病毒（HPV）检测呈阳性。此外，I型酪氨酸激酶受体ErbB - 2在至少30%的HNSCC和CxCa中过表达。最近，我们证明16型HPV的E6/E7与ErbB - 2协同作用可诱导人正常口腔上皮（NOE）细胞发生细胞转化。这伴随着NOE细胞中细胞周期蛋白D1的过表达。为了确定细胞周期蛋白D1在E6/E7/ErbB - 2协同作用中的作用，我们检测了E6/E7和ErbB - 2的独立作用，以及E6/E7和ErbB - 2在小鼠正常胚胎成纤维细胞（NEF）、野生型（wt）和细胞周期蛋白D1基因敲除（D1(-/-)）细胞中的联合作用。我们报告称，单独用E6/E7以及同时用E6/E7/ErbB - 2转导的NEF - wt细胞分别在裸鼠中形成小肿瘤和大肿瘤，并且在软琼脂中形成大小不同的集落；而单独的ErbB - 2在体内既不引发肿瘤形成，在软琼脂中也不形成集落。更重要的是，E6/E7、ErbB - 2以及E6/E7/ErbB - 2共同作用均无法在体内和体外诱导细胞周期蛋白D1(-/-)细胞发生肿瘤转化。此外，使用反义细胞周期蛋白D1，我们完全抑制了NEF - wt - E6/E7和wt - E6/E7 - ErbB - 2以及人NOE - E6/E7 - ErbB - 2转化细胞的肿瘤和集落形成。这些分析表明，细胞周期蛋白D1是正常细胞中由E6/E7或E6/E7/ErbB - 2协同作用诱导肿瘤转化的下游靶点。我们的数据表明，抗细胞周期蛋白D1疗法在治疗所有表达高危型HPV或HPV/ErbB - 2的人类癌症中可能具有高度特异性。
