沉默PHLDA1基因表达抑制胃癌细胞增殖

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、Cleaved-caspase-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。

Silencing of PHLDA1 expression inhibits proliferation of gastric cancer cells

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 PHLDA1), 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 PHLDA1 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. ［Key words］PHLDA1; gastric cancer; proliferation; apoptosis related protein