自分泌骨桥蛋白在泡状棘球蚴多功能干细胞迁移和转移中的作用研究

Alveoor echinococcosis (AE) is a potentially fatal zoonosis caused by Echinococcus multilocularis (E.m) , primary in the liver, showing the growth of maligant invasion, can secondary to lung and brain metastasis, the mechanism that invasive and metastasis in the host is not clear. The germinal layer cell is a kind of pluripotent stem cell, alveococcus new vesicle formation and growth, scolex occurrence and development, transmission and transfer of Echinococcus multilocularis in intermediate host depend on their self-renewal and differentiation ability. Osteopontin (OPN) is a kind of multifunctional cytokine and adhesion proteins, can promote cancer stem cell migration and metastasis. Our previous study found that OPN strong positive expression in rat liver alveolar hydatid cyst and metastatic lesions. So we propose an assumption that autocrine OPN of alveolar echinococcus germinal layer stem cell promotes invasive growth and metastasis. This study plan to obtain the germinal layer stem cell by culturing the cells in sphere medium, to detect the expression and correlation of OPN and Key stem cell factor. Exogenous administration of EGFR inhibitors to determine whether OPN regulates the development of the stem cell phenotype by EGFR mediated signaling pathway. Downregulate the OPN of germinal layer stem cell by RNA interference (RNAi) and construct nude mouse model, to observe the effect of OPN on alveoor echinococcosis growth and development. The aim of this study is to provide a new target and experimental basis for the treatment of the disease.

泡状棘球蚴病是一种危害极大的人兽共患寄生虫病，原发于肝脏，呈恶性浸润性生长，其浸润性生长和转移的机制尚不明确。泡球蚴生发层细胞是一种全能干细胞，泡球蚴新囊泡的形成和成长，头节的发生和发育以及泡球蚴在中间宿主体内的传播和转移，均以其自我更新和分化能力为基础。骨桥蛋白(OPN)是一种多功能的细胞因子和粘附蛋白，可促进肿瘤干细胞迁移和转移，前期研究发现，OPN在鼠肝泡球蚴和转移灶中强阳性表达，我们假设泡球蚴生发层干细胞自分泌OPN促进泡球蚴侵袭性生长和转移。本研究利用成球培养获得泡球蚴生发层干细胞，检测泡球蚴生发层干细胞OPN和关键干性因子表达及关系。外源性给予EGFR抑制剂,验证OPN是否通过EGFR介导的信号通路调控生发层干细胞表型；通过RNA干扰(RNAi)下调泡球蚴生发层干细胞OPN的表达并构建裸鼠模型，观察其在体外对泡球蚴生长的影响，为该病的药物治疗提供新的靶点及实验依据。

泡状棘球蚴病是一种危害极大的人兽共患寄生虫病，原发于肝脏，呈恶性浸润性生长，手术及药物治疗效果差，其在宿主体内浸润性生长和转移的机制尚不明确。骨桥蛋白(OPN)是一种多功能的细胞因子和粘附蛋白，可促进肿瘤干细胞迁移和转移，前期研究发现，OPN在鼠肝泡球蚴和转移灶中强阳性表达，我们假设泡球蚴原头节自分泌OPN促进泡球蚴侵袭性生长和转移。本研究克隆了泡球蚴OPN基因并研究分析其序列及蛋白，发现了泡球蚴原头节与肝癌细胞相互作用；建立泡状棘球蚴与肝细胞共培养模型，证实了泡球蚴原头节可促进细胞的增殖而抑制其凋亡；敲低 EmOPN 表达可显著抑制EGFR的磷酸化和多房棘球蚴的生长与肝外转移，阐明了多房棘球蚴自分泌的OPN可能通过EGFR通路参与其在肝内的生长、侵袭和肝外转移；通过对感染E.m患者的血清及手术标本进行OPN表达测定，发现OPN在感染E.m患者的血清及手术标本高表达，因此OPN可有望成为治疗AE的有效靶点，为后期临床试验提供帮助。