人乳头状瘤病毒16型与促癌物TPA协同作用诱发人胚口腔细胞恶性转化研究

Induction of human oral carcinoma by human papilomavirus 16 E6/E7 and TPA

目的 研究人乳头状瘤病毒 16型 (HPV16 )与TPA(12 O tetradecanog 1phorbol 13 acetate)协同作用在scid小鼠体内诱发人胚口腔细胞的恶性转化。方法 制备包装含有HPV 16E6 E7基因的逆转录病毒 ,用此病毒感染人胚口腔粘膜 ,将组织块接种于scid小鼠右侧肩部皮下 ,共分为 4组 :实验组为感染病毒的口腔粘膜和TPA ,共接种 8只小鼠 ;病毒组为感染病毒的口腔粘膜 ,共接种 6只小鼠 ;促癌组为正常口腔粘膜和TPA ,共接种 6只小鼠 ;对照组为正常口腔粘膜 ,共接种 5只小鼠。于接种第 3日起在左侧肩部皮下注射TPA ,每周一次。观察 16周后处死动物 ,对瘤组织进行病理诊断 ,并作聚合酶链反应 (PCR)检测HPV 16E6 E7基因。结果 实验组成瘤率为 7 8,其他 3组成瘤率皆为 0 6、0 6、0 5。实验组肿瘤组织的病理学检查结果证实为生长活跃的纤维组织细胞瘤 ,在肿瘤组织中用PCR检测到HPV 16E6 E7基因。结论 口腔细胞在感染含有HPV 16E6 E7基因的逆转录病毒后 。

Objective To study the effect of human papillomavirus (HPV) 16 E6/E7 and TPA (12 O tetradecanog 1 phorbol 13 acetate) on malignant transformation of human embryo oral tissue Methods Recombinant plasmid with HPV 16 E6/E7 was constructed and transfected into human embryo oral tissue The oral tissue with HPV 16 E6/E7 gene or without the gene was inoculated into the hypophloeodal of right shoulder in scid mice, respectively The study was conducted in four groups: the first group was the oral tissue transfected plasmid with HPV 16 E6/E7 plus TPA,which were inoculated into 8 scid mice; the second group was only oral tissue transfected with plasmid with HPV 16 E6/E7 into 6 scid mice; the third group was normal oral tissue plus TPA inoculated into 6 scid mice; and the final group was only normal oral tissue inoculated into 5 scid mice Three days after inoculation, TPA was injected at the left shoulder of the mice once a week Twelve weeks after inoculation, tumor was found in 7 scid mice from the first group HPV 16 E6/E7 gene in tumor tissues was analyzed by PCR Results The rate of tumor formation was 7/8 in the first group; no tumor was found in the other groups Pathological diagnosis of the tumor was fibrohistiocytoma HPV 16 E6/E7 gene was detected by PCR in tumor tissues Conclusion With the cooperating action of TPA, human oral tissue containing HPV 16 E6/E7 gene could cause malignant transformation in scid mice