沉默PHLDA1基因表达抑制胃癌细胞增殖

Silencing of PHLDA1 expression inhibits proliferation of gastric cancer cells

目的:明确普列克底物蛋白同源样结构域家族A成员1蛋白(PHLDA1)在各胃癌细胞系中的表达,探讨其对胃癌细胞生物学特性的影响。方法:先采用蛋白质印迹和qRT-PCR法检测MGC-803、BGC-823、MKN45、SGC-7901和HGC-27共5种胃癌细胞系中PHLDA1蛋白和mRNA的表达;再将MGC-803细胞分为对照组(转染乱序RNA)和干扰组(转染靶向干扰PHLDA1的siRNA),蛋白质印迹法检测2组细胞中PHLDA1蛋白的表达;CCK8实验检测细胞增殖率;平板克隆形成实验检测细胞克隆形成能力;蛋白质印迹检测各组细胞中Bax、Caspase-8、Cleaved-caspase-8、Caspase-3、Cleaved-caspase-3、Caspase-9和Cleaved-caspase-9凋亡相关蛋白的表达。结果:PHLDA1在5种胃癌细胞中呈不同程度高表达,在MGC-803细胞中表达量最高;与对照组比较,干扰组PHLDA1表达量显著降低(P<0.01),细胞增殖率明显减慢,克隆形成数显著减少、单个克隆的细胞数明显减少(P均<0.05),Cleaved-caspase-8、Cleavedd-caspasee-3、Caspase-9和Cleaved-caspase-9蛋白表达明显升高(P均<0.05)。结论:PHLDA1在胃癌细胞系中呈高表达,沉默其表达可抑制胃癌细胞增殖。

Objective： To investigate the expression of pleckstrin homology-like domain family A member 1（PHLDA1） in gastric cancer cell lines and its effect on the biological characteristics of gastric cancer cells. Methods： Western blotting and qRT-PCR were used to detect the expression of PHLDA1 in gastric cancer cell lines MGC-803, BGC-823, MKN45, SGC-7901 and HGC-27. MGC-803 cells were divided into control group（transfected with scrambled RNA） and interference group（transfected with siRNA targeting PHLDAl）, Western blotting was used to detect the expression of PHLDA1;CCK8 assay was used to test the cell proliferation rate; plate colony formation assay was used to detect the colony formation ability; Western blotting was used to detect expression of apoptosis-related proteins such as Bax,Caspase-8,Cleaved-caspase-8,Caspase-3,Cleaved-caspase-3,Caspase-9,and Cleaved-caspase-9 in each group of cells. Results ： PHLDA1 was highly expressed in five gastric cancer cell lines at different degree,among which the PHLDAI expression was the highest in MGC-803 cells. Compared with the control group, the interference group showed lower expression of PHLDA1（P〈0.01）, and less cell proliferation rate, smaller number of cell clones and single cloned cells（P〈0. 05）, and higher expression of Cleaved caspase-8,Cleaved-caspase-3,Caspase-9,and Cleaved-caspase-9 proteins（P〈0. 05）. Conclusion： PHLDA1 was highly expressed in gastric cancer cell lines. Silencing PHLDA1 expression could inhibit proliferation of gastric cancer cell.