HPV持续感染与宫颈癌特异性改变的共性蛋白质表达调控网络研究

Cervical cancer has been one the high-incident diseases in Xinjiang. In previous works, we have preliminarily understood the mechanisms and early prognostic markers of cervical cancer in Uighur women at systems biology level by a series of studies including serum proteomics, metabonomics, and epigenetics. However, many questions arised about the cellular regulation network of protein expression in cervical cancer tissues, which was poorly described in the literature. The infection by human papillomaviruses (HPV) especially the persistent infection by high-risk type HPV16 may be the main etiological factor responsibe for cervical carcinogenesis, but there is a relatively long latent period for cervical precursor development after virus infection, thus HPV screening has a very restrictive potential in cancer prognosis.In this project, we will focus on Uighur women with cervical carcinoma and analyze HPV16-positive cervical lesions and identify candidate differentially expressed proteins followed by quantitative detection, by a proteomics approach using technological platforms including two-dimensional liquid-phase chromatography, two-directional fluorescence in-gel electrophoresis and mass spectrometry etc., study the regulation network of protein expression altered by, and specific to, persistent HPV infection and cervical carcinogenesis,for understanding the mechanisms of cervical carcinogenesis common to tumors and the characteristics unique to the ethnic population，and establishing an auxiliary diagnostic profile for early prognosis of cervical cancer in addition to HPV detection.This study is important to improve the management quality of cervical cancer prevention and treatment in Xinjiang, and promote multidisciplinary development as well.

宫颈癌是新疆特高发疾病之一。前期工作中，通过维吾尔族妇女宫颈癌的血清蛋白质组学、代谢组学和表观遗传学等多方面研究，我们从系统生物学水平初步认识宫颈癌发病机制和早期预警指标。但是，很多问题直接涉及宫颈癌组织细胞内蛋白质表达调控网络，而与此相关的文献甚少。人乳头瘤病毒（human papillomavirus, HPV）感染，尤其是高危型HPV16持续感染可能是宫颈癌发生的主要原因，但是病毒感染与癌前病变的潜伏期较长，HPV筛查预警潜力极其有限。故本课题拟利用维吾尔族妇女宫颈癌疾病资源,依托二维液相色谱、双向荧光差异电泳和质谱等技术平台，通过HPV16阳性宫颈病变组织的蛋白质组学分析及候选差异表达蛋白的发现和定量鉴定，探讨HPV持续感染及宫颈癌特异性改变的共性蛋白质表达调控网络，了解宫颈癌发病机制的肿瘤共性和族群特点，建立基于HPV筛查的宫颈癌预警辅助诊断指标，提高新疆宫颈癌防治水平，并促进多学科发展。

维吾尔族女性宫颈癌是新疆高发肿瘤，其早期发现是有效治疗的关键。人乳头瘤病毒（human papillomavirus, HPV）感染，尤其是高危型HPV16持续感染是宫颈癌发生的主要因素。以往工作中，我们通过在明确宫颈鳞癌发生与多种血浆蛋白质和小分子代谢物含量变化有一定的因果关系。但是，一系列问题直接涉及宫颈癌组织内蛋白质表达调控网络。故本课题以汉族和维吾尔族女性宫颈鳞癌患者（临床分期Ia 至 IIa）为研究对象，选择基于iTRAQ（同重标签相对和绝对定量）技术的蛋白质组学方法，对HPV16阳性宫颈鳞癌和HPV阴性正常对照组织进行蛋白质表达谱比较分析，鉴别出67种差异表达蛋白质；根据生物信息学和查新分析结果，宫颈癌发生及HPV感染与大部分候选蛋白质表达调控的关系研究尚未见报道；以此为基础，选择24种候选蛋白质指标，对76例宫颈癌及癌前病变（CIN Ⅱ-Ⅲ）组织进行了定量RT-PCR分析，明确宫颈鳞癌发生伴有23种候选基因的mRNA表达水平显著差异，其变化趋势与蛋白质组学数据一致，验证了高通量分析结果的特异性和准确度；同时，选择7种候选蛋白质指标，对116例石蜡包埋组织切片进行了免疫组织化学分析，对宫颈鳞癌与癌前病变（CIN Ⅱ-Ⅲ）及正常对照相比较，明确所有蛋白质指标均有显著差异，进一步支持蛋白质组学和定量RT-PCR分析结果。由于宫颈癌及癌前病变组织的高危型HPV检出率很高，无法从临床组织标本水平上，评价HPV感染与单一候选蛋白质表达水平的关系，从而启动了细胞水平的研究工作。通过本研究，明确宫颈癌发生及HPV16感染与肿瘤组织内整体水平的蛋白质表达调控存在密切关系，并为基于HPV筛查的宫颈癌预警提供了一系列候选辅助诊断指标；发表了11篇科研论文，探讨了宫颈癌发病机制及早期预警，产生了一定的学术影响；培养了4名博士和硕士学位研究生；3名课题组成员申报与获得省级自然科学基金资助，为继续开展本课题后续研究奠定了基础。