DMBT1过表达对胆囊癌细胞系GBC-SD粘附增殖的影响

目的:观察DMBT1体外过表达对胆囊癌细胞同质、异质粘附能力及细胞增殖能力的影响。方法:以脂质体为介导在体外建立稳定表达DMBT1的胆囊癌细胞系,细胞分组:GBC-SD/DMBT1,转染DMBT1组;GBC-SD/Vector,转染空载体组;GBC-SD,未处理组;平板克隆形成实验检测细胞集落形成能力,细胞聚集实验观察细胞同质粘附能力,层粘蛋白实验及人脐静脉上皮细胞(HUVEC)粘附实验观察细胞异质粘附能力。统计学采用配对或独立样本t检验,P<0.05有统计学意义。结果:平板克隆实验发现GBC-SD/DMBT1组克隆形成数20±3,GBC-SD/Vector组46±8,差异有统计学意义(P<0.01);细胞聚集实验表明GBC-SD/DMBT1聚集指数在振摇时间40、60min时分别为0.471±0.041、0.610±0.050,显著高于GBC-SD/Vector(40、60min时分别为0.250±0.024、0.312±0.026),差异有统计学意义(P<0.01);层粘蛋白Ln粘附实验表明GBC-SD/DMBT1在培养60、90min时残留细胞吸光度分别为0.157±0.008及0.139±0.010显著低于GBC-SD/Vector(60、90min时分别为0.347±0.026、0.476±0.023),差异有统计学意义(P<0.01);人脐静脉粘附表明GBC-SD/DMBT1粘附率为0.341±0.042、GBC-SD/vector粘附率为0.694±0.058,差异有统计学意义(P<0.01)。结论:DMBT1在体外过表达抑制胆囊癌细胞的增殖能力,促进胆囊癌细胞与细胞之间的粘附,抑制细胞与基质或异种细胞的粘附。

Effects of DMBT1 over-expression in gallbladder carcinoma cell line GBC-SD on its adhesion and proliferation

Objective:To observe the effects of DMBT1 over-expression in gallbladder carcinoma cell line GBC-SD on its adhesion and proliferation.Methods:The cell model with stable expression of DMBT1 has been successfully established in vitro.Flat plate clone formation assay was employed to assess its proliferation ability.Cell aggregation assay was applied to evaluate alterations of homogeneous adhesion ability,whereas laminin(Ln) adhesion assay and human umbilical vein endothelial cell (HUEC) adhesion assay were used to assess its heterogeneous adhesion ability.Results:Flat plate clone formation assay demonstrated more clones in GBC-SD/DMBT1(20±3)(P<0.01),versus those in GBC-SD/Vector(46±8).Cell aggregation assay witnessed markedly higher aggregation index of GBC-SD/DMBT1 (P<0.01),versus that of GBC-SD/vector when incubation time was 40 min and 60 min.Ln adhesion assay further demonstrated a decrease of Ln adhesion ability of GBC-SD/DMBT1(P<0.01),versus GBC-SD/vector after 60min and 90min of interaction.Furthermore,HUEC adhesion assay indicated that adherence ratio of GBC-SD/DMBT1 was significantly lower than that of GBC-SD/Vector (P<0.01).Conclusion:DMBT1 over-expression in vitro promotes homogeneous adhesion yet inhibited heterogeneous adhesion of GBC-SD and its proliferation.