小干扰RNA沉默半乳糖凝集素3基因表达对胆管癌细胞增殖凋亡的影响及机制  

Effects and its mechanism of silencing Galectin- 3 gene expression by small interfering RNA on proliferation and apoptosis of cholangiocarcinoma cells

作　　者：苏智雄[1] 叶新平[2] 尚丽明[2] 

机构地区：[1]广西医科大学第二附属医院普通外科,南宁530007 [2]广西医科大学第一附属医院肝胆外科,南宁530021

出　　处：《中华实验外科杂志》2018年第1期79-81,共3页Chinese Journal of Experimental Surgery

摘　　要：目的抑制半乳糖凝集素3（Galectin-3）基因表达对胆管癌细胞增殖凋亡的影响及机制。方法反转录-聚合酶链反应（RT-PCR）和Western blot检测胆管癌组织中Galectin-3基因的mRNA及蛋白表达；si-Galectin-3转染HCCC-9801细胞，同时转染阴性对照组和正常对照组，转染48 h后Western blot检测Galectin-3、细胞核增殖抗原（Ki-67）、活化的含半胱氨酸的天冬氨酸蛋白水解酶3（Cleaved Caspase-3）、β-连环蛋白（β-catenin）、细胞周期蛋白D1（Cyclin D1）蛋白表达；细胞计数试剂盒（CCK-8）实验和流式细胞仪分别检测细胞增殖及凋亡。结果胆管癌组织中Galectin-3基因的mRNA（5.558±0.056）及蛋白表达（0.269±0.025）显著高于癌旁组织（0.977±0.031、0.037±0.011）（tmRNA＝8.626，PmRNA＝0.003；t蛋白表达＝8.997，P蛋白表达＝0.002）；si-Galectin-3组Galectin-3（0.094±0.012）、Ki-67（0.201±0.025）、β-catenin（0.261±0.024）、Cyclin D1（0.137±0.017）蛋白表达及细胞存活率[（95.26±3.42）%]均显著低于Normal组[（0.358±0.031）%、（0.426±0.032）%、（0.894±0.047）%、（0.435±0.031）%、（71.12±4.48）%]（tGalectin-3＝9.007，PGalectin-3＝0.002；tKi-67＝8.138，PKi-67＝0.003；tβ-catenin＝9.345，Pβ-catenin＝0.002；tCyclin D1＝9.977，PCyclin D1＝0.001；t细胞存活率＝8.452，P细胞存活率＝0.003）；细胞凋亡率[（17.12±1.28）%]及Cleaved Caspase-3（0.121±0.015）表达均显著高于对照组[（1.54±0.47）%、0.031±0.011]（t细胞凋亡率＝10.002，P细胞凋亡率＝0.001；tCleaved Caspase-3＝9.879，PCleaved Caspase-3＝0.001）。结论下调Galectin-3基因表达可通过抑制Wnt/β-catenin信号通路降低HCCC-9801细胞增殖及诱导细胞的凋亡。Objective To study the effect and mechanism of inhibition of Galectin-3 gene expression on proliferation and apoptosis of cholangiocarcinoma cells.MethodsThe mRNA and protein expression of Galectin-3 gene in cholangiocarcinoma were detected by reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting; small interfering RNA （siRNA）-Galectin-3 was transfected into HCCC-9801 cells, and cells transfected with negative control group and normal control group, the expression of Galectin-3, Ki-67, Cleaved Caspase-3, β-catenin, Cyclin D1 protein were detected by Western blotting after 48 h; CCK-8 assay and flow cytometry were used to detect the proliferation and apoptosis of cell.ResultsThe mRNA （5.558±0.056）and protein （0.269±0.025）expression of Galectin-3 gene in cholangiocarcinoma was significantly higher than that in adjacent tissues （0.977±0.031） and （0.037±0.011） （tmRNA=8.626, PmRNA=0.003; tProtein expression=8.997, PProtein expression=0.002）; protein expression of Galectin-3 （0.094±0.012）, Ki-67（0.201±0.025）, β-catenin （0.261±0.024）, Cyclin D1 （0.137±0.017） and cell survival rate （95.26±3.42）% in si-Galectin-3 group was significantly lower than that of Normal group [（0.358±0.031）%, （0.426±0.032）%, （0.894±0.047）%, （0.435±0.031）%, （71.12±4.48）%] （tGalectin-3=9.007, PGalectin-3=0.002; tKi-67=8.138, PKi-67=0.003; tβ-catenin=9.345, Pβ-catenin=0.002; tCyclin D1=9.977, PCyclin D1=0.001; tcell survival rate=8.452, Pcell survival rate=0.003）; the apoptosis rate （17.12±1.28）% and Cleaved Caspase-3 （0.121±0.015） protein expression was significantly higher than the control group [（1.54±0.47）%, 0.031±0.011] （tapoptosis rate=10.002, Papoptosis rate=0.001; tCleaved Caspase-3=9.879, PCleaved Caspase-3=0.001）.ConclusionDown regulation of Galectin-3 gene expression can inhibit the proliferation of HCCC-9801 cells and induce cell apoptosis by inhibiting the Wnt/β-catenin signaling pathway.

关 键 词：半乳糖凝集素3基因 胆管癌 增殖 凋亡 

分 类 号：R735.8[医药卫生—肿瘤]