基于Wnt/β-catenin 通路GPC3促异质卵圆细胞亚群恶变机制研究

Heterogeneous Oval cells were probably the precursor cells of hepatocellular carcinoma due to literatures. Persisting expression of memberane protein glypican 3 (GPC3) in activated oval cells could transform the oval cells to cancer cells, furthermore GPC3 could also stimulate Wnt/β-catenin pathway to make hepatoma cells proliferation. In previous research we have identified the oval cell subset (GPC3+) origined from 2AAF/PH SD rat model, cloned rat GPC3 gene and constructed two recombinated plasmids containing full lenghth or truncated GPC gene. In order to elucidate the effect of GPC3 promoting oval cell canceration and illustrate the relationship between the effect with Wnt/β-catenin pathway, three plasmids including pcDNA3.1(+)/GPC3-ORF, pcDNA3.1(+)/GPC3-550C (without C-terninal) and si RNA-GPC3 were transfected into the oval cell subset and established the over-expression, non-expression of GPC3-ORF or truncated GPC3 cell clones. In vitro the key elements of Wnt/β-catenin pathway and the factors related canceration were detected.

异质卵圆细胞很可能是肝癌前体细胞，GPC3 在卵圆细胞持续表达可使之恶变，又经刺激Wnt/β -catenin 通路促肝癌细胞生长。前期研究我们已鉴定2AAF/PH SD 大鼠模型GPC3+卵圆细胞亚群，克隆大鼠GPC3 基因、构建全长和截短型GPC3 质粒。为明晰GPC3促卵圆细胞癌变作用，阐明其与Wnt/β -catenin 通路关系，再构建si-RNA GPC3 质粒，三者转染该亚群细胞，建立过表达、无表达全长和截短GPC3 细胞克隆，体外检测Wnt/β-catenin 通路中关键因子以及细胞癌变分子；体内检测裸鼠荷瘤模型成瘤及生存状况；以期从体外、体内多个水平验证基于Wnt/β-catenin 通路GPC3促异质卵圆细胞亚群恶变假说，为阻断损伤肝癌变提供治疗靶点。