PHF5A通过NF-κB通路影响胃癌细胞MGC803的增殖和迁移  

Effect of PHF5A on proliferation and migration of human gastric cancer cells via regulating NF-κB signaling

作　　者：曹一凡 张如通 苟雅雯 陈乐乐 陈思 张开光 Cao Yifan;Zhang Rutong;Gou Yawen(Dept of Gastroenterology,The Afiliated Provincial Hospial of Anhui Medical University,Hefei 230001)

机构地区：[1]安徽医科大学附属省立医院消化内科,合肥230001

出　　处：《安徽医科大学学报》2020年第8期1198-1203,共6页Acta Universitatis Medicinalis Anhui

基　　金：安徽省科技计划项目(编号:1604a0802075)。

摘　　要：目的观察PHD锌指结构域蛋白5A(PHF5A)基因在胃癌细胞和胃正常细胞中的表达情况,阐明敲低PHF5A基因可能通过影响NF-κB通路抑制胃癌细胞的增殖、克隆形成和迁移。方法使用Western blot和qPCR法检测PHF5A在胃正常上皮细胞GES-1和胃癌细胞系MGC803、MKN45、AGS中的表达情况。设置转染PLKO.1空载质粒的MGC803细胞为阴性对照,转染PLKO.1-ShPHF5A-Sh1和PLKO.1-ShPHF5A-Sh2质粒的作为实验组,使用两组不同序列在MGC803中抑制PHF5A的表达。采用克隆形成、细胞增殖计数检测细胞增殖情况,采用划痕实验检测细胞体外迁移能力变化。用Western blot检测阴性对照组和实验组中NF-κB P65(核内)、p-IkBα,IkBα,Cyclin D1蛋白表达情况。结果Western blot和qPCR结果显示胃癌细胞系MGC803、AGS、MKN45中PHF5A的表达量高于胃正常细胞系GES-1,差异有统计学意义。在MGC803细胞中抑制PHF5A的表达后,PHF5A蛋白的表达水平降低。与对照组比较,细胞的增殖和迁移都受到了抑制。同时,实验组细胞NF-κB P65核内表达降低,IκBα磷酸化水平下降,Cyclin D1表达水平下降(P<0.05)。结论PHF5A在胃癌肿瘤细胞中的表达显示高于正常细胞,可能通过NF-κB信号通路来影响胃癌细胞MGC803的增殖和迁移。Objective To investigate the expression and effects of PHD finger-like domain-containing protein 5 A(PHF5 A)in gastric cancer cells and to explore its mechanism.Methods QPT-PCR and Western blot were used to detect the expression levels of PHF5 A mRNA and protein in normal gastric cancer cell line GES-1 and gastric cancer cell lines MKN45,MGC803 and AGS.MGC803 cells transfected with negative control empty lentivirus were used as the control group.Two knockdown groups which were transfected with two different interfering sequences lentivirus against the PHF5 A gene were used as the experimental groups.The proliferation ability was observed by cell proliferation curves and colony formation assay.The migration ability was observed by wound healing assay.Western blot was used to detect the expression levels of some signaling molecules such as Transcription factor p65(NF-κB p65),NF-kappa-B inhibitor alpha(Iκbα),p-Iκbαand Cyclin-D1-binding protein(Cyclin D1)in NF-κB signal transduction pathway.Result The expression levels of PHF5 A mRNA and protein in gastric cancer cells were significantly higher than those in normal gastric mucosal epithelial cells.After knockdown the expression of PHF5 A gene,the level of PHF5 A protein decreased.Compared with the control group,the proliferation,migration and colony formation ability of MGC803 cells in two PHF5 A knockdown groups sh1 and sh2 decreased obviously.At the same time,the result of western blot showed that the expression of NF-κB,p-IκBα,Cyclin D1 significantly decreased in experiment groups(P<0.05).Conclusion PHF5 A is highly expressed in gastric cancer cells.Lentivirus-mediated inhibition of PHF5 A gene expression can reduce the proliferation and migration ability of MGC803 cells.The mechanism may be related to the inhibition of NF-κB signaling pathway.

关 键 词：胃癌 PHF5A NF-ΚB信号通路 MGC803 

分 类 号：R735.2[医药卫生—肿瘤]