雄黄对高危型HPV感染的宫颈上皮内瘤变中PI3K-Akt信号通路的干预研究

Persistent expression of the viral E6/E7 oncogenes of high-risk human papillomaviruses (HR-HPV) acts as a key factor in making precancerous lesions progress into invasive cervical cancer. Currently, on the CIN treatment traditional surgery is the major choice, but there are not non-invasive and effective blocking methods. In the previous studies a traditional Chinese medicine prescription containing realgar (AS4S4) was found to cut off HPV infection and promote the healing of CIN with positive HR-HPV by inducing CIN cell apoptosis and down-regulating PI3K、p-Akt and E6/E7 protein. PI3K-Akt signaling pathway plays an important role in regulating apoptosis and maintaining process of consistant HPV infection. The project will observe the interversion effect of AS4S4 on PI3K-Akt pathway in HPV16 positive CIN cells model and HPV transgenic mouse model and detect the impact of AS4S4 on PI3K-Akt pathway and its downstream signaling molecules by using molecular biologic technology to activate or inhibit the PI3K-Akt pathway. Expliciting the interversion role of AS4S4 on PI3K-Akt pathway and uncovering the molecular mechanism of AS4S4 clearing HPV infection and reversing CIN disease would provide the theoretical and scientific basis for AS4S4 effectively treating CIN.

高危型人乳头瘤病毒（HR-HPV）E6/E7癌基因持续表达是宫颈癌前病变（CIN）进展为宫颈癌的关键因素。目前治疗CIN仍以手术为主，尚缺乏无创、特效的阻断方法。本课题组前期研究发现含雄黄中药验方可阻断HPV感染，促进HR-HPV阳性CIN愈合。其君药雄黄（As4S4）可诱导CIN细胞凋亡，下调PI3K、p-Akt及HPV16E6/E7表达。PI3K-Akt通路在调节细胞凋亡，维持HPV感染过程中发挥重要作用。因此，本项目拟在HPV16阳性CIN细胞和HPV转基因小鼠模型中观察AS4S4对PI3K-Akt通路的干预作用，采用分子生物学手段，激活或抑制PI3K-Akt通路，检测AS4S4对PI3K-Akt通路及其下游信号分子的影响。明确AS4S4对PI3K-Akt通路的干预作用，阐明AS4S4清除HPV、逆转CIN的作用机制，为AS4S4治疗CIN提供理论基础和科学依据。

雄黄的主要成分是AS4S4。在前期研究发现As4S4可诱导宫颈癌细胞凋亡，下调PI3K、AKT及HPV16E6/E7表达的基础上，我们进一步观察了AS4S4对HPV阳性细胞增殖、侵袭的抑制作用并研究了其作用机制；同时，对以“雄黄”为君药的中药复方促高危型HPV转阴、逆转宫颈病变的作用机制进行了研究。本课题发现As4S4对HPV阳性细胞具有浓度依赖性的生长抑制和诱导凋亡作用；As4S4通过调控P53的表达从而引起Bcl-2/Bax、Caspase3等凋亡相关基因的调变。另外，我们用以“雄黄”为君药的中药复方治疗高危型HPV持续感染的宫颈病变，应用基因芯片技术筛选了宫颈组织在药物治疗前后差异表达的基因，验证了药物显著抑制PI3K-AKT信号通路外，还对吞噬体以及免疫相关信号通路的作用也尤为明显，这为我们进一步深入的研究雄黄治疗高危型HPV感染提供了理论基础和科学依据。