骨桥蛋白介导的卵圆细胞增殖在乙肝相关性肝细胞肝癌侵袭转移中的作用及机制研究

Abstract:Although the treatment of HCC has been markedly improved, its recurrence and metastasis still are key causes of HCC bad prognosis. It is estimated that the liver cancer stem cell contributes to HCC recurrence and metastasis. Oval cell is regard as liver cancer stem cell because of its activated proliferation resulting in carcinogenesis of HCC. It was found that OPN expression was higher in the HBV-related HCC group than the HCC group without HBV infection. HBX enhanced the transcriptional activity of OPN promoter and OPN promoted the proliferation of hepatic oval cells. Therefore, we propose the hypothesis which HBX regulating OPN expression may cause the activation, over proliferation and malignant transformation of hepatic oval cells link to the recurrence and metastasis of HCC. This study was designed that IE/6 oval cell line was used as an experimental model and bioinformatics, biochemistry, molecular biology, pathology were applied as technical supports. The role of OPN will be confirmed in the activation and over proliferation of hepatic oval cells. This study will further clarify that HBX regulating-OPN plays a role in malignant transformation of hepatic oval cells and a significance in the recurrence and metastasis of HCC. The prospective results will provide new idea and target spot of prevention and cure of recurrence and metastasis of HCC postoperation.

肝细胞癌（HCC）术后的侵袭转移是肝癌高复发率的重要原因，癌干细胞可能在肝癌细胞增殖、侵袭转移中发挥重要作用。近期发现，肝癌干细胞具有较强的迁移、侵袭能力，可促进肝癌的转移，但机制不清。卵圆细胞是肝癌干细胞。骨桥蛋白（OPN）与肝癌的侵袭转移密切相关。我们前期研究发现，OPN可促进卵圆细胞的增殖并在乙肝相关性HCC中高表达；乙肝X蛋白可激活OPN启动子的转录活性。本课题将采用基因转染技术观察HBx对OPN基因表达的影响，探讨HBx对骨桥蛋白表达及卵圆细胞增殖侵袭的影响机制。明确OPN/卵圆细胞途径在肝癌侵袭转移中的作用，对肝癌术后侵袭转移的防治具有重要意义。

摘要：肝细胞癌术后的侵袭转移是肝癌高复发率的重要原因，癌干细胞可能在肝癌细胞增殖、侵袭转移中发挥重要作用。我们的研究，通过对肝卵圆细胞LE/6培养，构建OPN启动子区不同长度的DNA片段并将其克隆到荧光素酶报告基因载体上，构建一系列OPN启动子区不同长度的质粒。将X蛋白表达载体分别与上述OPN启动子区各片段质粒共转染，通过检测荧光素酶活性，发现加入HBx的实验组荧光素酶活性与未加入HBx的对照组差异显著，呈现明显的抑制作用，证实HBx对OPN启动子具有调控作用。接着分析发现整片段与opn-541片段差异显著，可以认为HBx对OPN启动子的调控区域在opn-541片段的可能性很大。通过对OPN启动子的调控区域可能存在的转录因子进行预测，然后经过缺失或者突变，观察其活性是否明显改变，初步确定出参与转录调控的可能转录因子结合位点。最后希望能明确OPN/卵圆细胞途径在肝癌侵袭转移中的作用，对肝癌术后侵袭转移的防治具有重要意义，为HCC转移复发的临床防治提供理论依据。