叉头框蛋白O3在胰腺癌中的表达及其对胰腺癌细胞的影响

The expression of FOXO3 in pancreatic cancer and its effects on pancreatic cancer cells

目的:分析叉头框蛋白O3(FOXO3)在胰腺癌中的表达及其对胰腺癌细胞运动和增殖的影响。方法:检索LinkedOmics数据库胰腺癌患者胰腺癌和癌旁组织FOXO3表达。Western印迹和实时定量聚合酶链反应检测胰腺癌细胞系和人胰腺星状细胞HPSC中FOXO3表达。选择低表达FOXO3的PANC-1、MIAPaCa-2胰腺癌细胞,分别转染FOXO3过表达质粒和阴性对照质粒。克隆形成实验、细胞划痕实验、Transwell侵袭实验和细胞周期检测各组胰腺癌细胞运动和增殖能力。结果:LinkedOmics数据库中,64例胰腺癌患者癌组织中FOXO3蛋白相对表达水平明显低于癌旁组织(t=8.36,P<0.001)。PANC-1细胞过表达FOXO3后细胞克隆数为(30.0±6.6)个(40倍视野下计数),低于阴性对照细胞的(92.7±6.7)个,差异有统计学意义(t=11.54,P<0.001)。PANC-1和MIAPaCa-2在上调FOXO3表达后细胞划痕修复率较对照组显著下降。Transwell实验PANC-1细胞FOXO3过表达后(每个100倍视野下)穿膜数(21.0±6.6)个,低于转染阴性对照质粒细胞的(55.7±8.5)个,差异具有统计学意义(t=5.59,P=0.005)。MIAPaCa-2细胞检测结果与PANC-1基本一致。PANC-1和MIAPaCa-2在过表达FOXO3后,细胞在G0/G1期的比例下降,而在S期比例增加。结论:FOXO3在胰腺癌中表达降低,上调FOXO3表达后能够显著抑制胰腺癌细胞的增殖、迁移、侵袭能力,还使细胞周期停滞在S期。FOXO3是治疗胰腺癌的潜在靶点。

Objective To investigate the expression of forkhead box protein O3(FOXO3)in pancreatic cancer and its effect on the motility and proliferation of pancreatic cancer cells.Methods The FOXO3 expression in pancreatic cancer and adjacent tissues was retrieved from LinkedOmics database.Western blotting and real-time quantitative polymerase chain reaction were used to detect FOXO3 expression in pancreatic cancer cells and human pancreatic stellate cells.PANC-1 and MIAPaCa-2 pancreatic cancer cells with low FOXO3 expression were selected to transfect FOXO3 overexpression plasmid and negative control plasmid,respectively.The motility and proliferation ability of pancreatic cancer cells were detected by colony formation assay,cell scratch assay,Transwell assay and flow cytometry.Results In the LinkedOmics database,the relative expression of FOXO3 protein in the cancer tissues of 64 patients with pancreatic cancer was significantly lower than that in the adjacent tissues(t=8.36,P<0.001).The number of clones in PANC-1 cell line was(30.0±6.6)after overexpressed FOXO3,which was lower than that in negative control cells(92.7±6.7),and the difference was statistically significant(t=11.54,P<0.001).After overexpressed FOXO3 in PANC-1 and MIAPaCa-2 cell lines,the scratch repair rate was significantly decreased compared with the control group.In Transwell experiment,the number of cells in FOXO3 overexpressed group in PANC-1 cell lines was(21.0±6.6),which was lower than that of negative control groups(55.7±8.5),and the difference was statistically significant(t=5.59,P=0.005).The results of MIAPaCa-2 cell line were consistent with that of PANC-1 cell line.After overexpressing of FOXO3 in PANC-1 and MIAPaCa-2 cell lines,the proportion of cells in the G0/G1 phase decreased,while the proportion in the S phase increased.Conclusion The expression of FOXO3 was decreased in pancreatic cancer.Overexpression of FOXO3 could significantly inhibit the proliferation,migration and invasion of pancreatic cancer cells and induce cell cycle arrest,which is a potential target for the treatment of pancreatic cancer.