【作者】 周晓波；

【导师】 徐宁志；

【作者基本信息】 中国协和医科大学 ， 细胞生物学， 2003， 博士

【摘要】 恶性肿瘤是严重危害人类健康的疾病之一，虽然经过各国科学家多年的努力，取得了许多可喜的成绩，但许多肿瘤的发病原因与发病机制至今仍然没有确切的答案。可以肯定的是，肿瘤的发生是一个多因素作用、多基因参与、经过多个阶段才最终形成的极其复杂的生物学过程。而感染因素作为肿瘤发生过程中一个比较重要的因素，越来越多地受到了人们的关注。人乳头瘤病毒(human papillomavirus，HPV)就是一类与多种肿瘤发生相关的感染因素。它与宫颈癌因果关系的确定大大推动了宫颈癌预防与治疗的进展，使得宫颈癌成为最有希望通过疫苗接种而得到控制的第一个人类恶性肿瘤。 除宫颈癌之外，HPV感染还可能与头颈部肿瘤及食管癌等其他肿瘤的发生相关，如果这些肿瘤与HPV的关系得到进一步确证，将会大大扩大HPV疫苗对肿瘤的有效预防范围，而食管癌又是我国的高发肿瘤之一，因此，很有必要对二者相关性作深入的研究。 为进一步确定HPV在我国食管癌中的作用，我们选取来自中国两个不同高发区(安阳与山西)的食管癌标本48例以及医科院肿瘤医院的食管癌30例(代表非高发区的群体)作为研究对象。针对HPV16 E6基因及其产物作检测，用PCR，原位杂交，免疫组化三种方法从DNA，mRNA，蛋白三个水平检测病毒感染及表达状况。研究对象包括78例食管鳞癌组织与23例癌旁“正常”食管上皮组织。结果显示：首先，HPV感染率具有明显的地域性差异。HPV感染率依次为安阳最高，其次为山西，而北京的最低。北京收诊病人来自全国各地，总体上代表了来自非高发区的病人群体，免疫组化方法检测仅有一例病毒癌蛋白E6阳性。其次，从三种方法的分析来看，来自安阳与山西的食管癌中有65％PCR阳性；而且，在PCR检测了HPV DNA基础之上，病毒DNA阳性的病例中大约三分之二被原位杂交证实为病毒的mRNA也为阳性。而病毒的E6蛋白的阳性率经免疫组化证实并没有其mRNA表达频率那么高，但也占出现了HPV16感染的肿瘤组织的40％(13／31)。第三，在检测癌旁所谓“正常”上皮细胞时，也可检出HPV病毒，说明病理上诊断为形态正常的上皮很有可能已经发生了微观的病变。 然后，我们对HPV16重要的癌蛋白之一，E7的作用机制进行探讨，主要从引起更多还原

【Abstract】 The malignant tumor is one of the most serious diseases that endangering people’s health. After years of hard work, the scientists have achieved a great number of breakthroughs in the basic research of carcinogenesis, while the exact reasons and mechanism of tumors are not completely known. But it is sure that the development of tumor is a very complex biological process with multifactor and many genes involved. And the virus infection as one of the most important factors among these is attracting more and more attention upon it. Human Papillomavirus (HPV) is a good example, which is believed to relate with many kinds of human tumors. After HPV was confirmed as the definite cause for cervical cancer, the prevention and therapy of cervical cancer were highly pushed forward. So the cervical cancer would possibly become the first tumor that could be controlled completely by vaccination.Besides cervical cancer, the HPV infection also related with head and neck cancer, esophageal cancer and so on. If the relationship between HPV and other tumors was confirmed, the application of HPV vaccine will be expanded. While the incidence of esophageal cancer in China is high, it would be beneficial to study the relationship between HPV and Chinese esophageal cancers further though it has been a controversial question for long time.To investigate the putative role of human papillomavirus (HPV) infection in the carcinogenesis of esophageal cancers in China, we studied the infection rate of HPV in 78 esophageal cancer cases from North China. Among them 48 cases were from high-incidence regions (Anyang city and Shanxi province) and 30 from non-high incidence region (Beijing). And the 23 cancers from Shanxi had the corresponded paracancer epithelium as normal control. All the samples were detected for the existence of HPV 16 DNA by PCR, mRNA in situ hybridization (ISH) and immunohistochemistry (IHC) targeting HPV16 E6 gene. As a result, the infection rate varied with different sample sources: Anyang had the highest HPV16 positive rate, with Shanxin in the middle, Beijing the least. From these two high-incidence regions, there更多还原