Expression of anti-silencing functional protein 1B in human gastric cancer cells infected with Helicobacter pylori and its effect on proliferation and metastasis of AGS cell line
Expression of anti-silencing functional protein 1B in human gastric cancer cells infected with Helicobacter pylori and its effect on proliferation and metastasis of AGS cell line
Objective To detect the expression level of histone chaperone anti-silencing functional protein 1B(ASF1B)in gastric cancer(GC)tissues and its relationship with the prognosis of GC patients.To investigate the effects of ASF1B on the proliferation and metastasis ability of human GC cell line AGS and the possible molecular mechanism.To analyze the relationship between Helicobacter pylori and the expression of ASF1B.Method The expression level of ASF1B in various of tumor tissues and adjacent normal tissues in the cancer genome atlas database was analyzed,and then the mRNA expression level of ASF1B in 59 pairs of GC tissues and paired adjacent normal tissues of our cohort were measured via real time fluorescent quantitative polymerase chain reaction(RT-qPCR).The relationship between the expression levels of ASF1B and the prognosis of GC patients in Kaplan-Meier plotter database was analyzed.Besides,the expression levels of ASF1B in GC tissue microarray were detected by immunohistochemistry,and then the relationship between ASF1B and clinical prognosis and clinicopathological parameters of GC patients was analyzed.Small interfering RNA(siRNA)technology was used to knockdown the expression of ASF1B in GC cell line AGS,and AGS cells were divided into siNC group(negative control transfected cells),siASF1B#1 group,siASF1B#2 group and siASF1B#3 group.Then the proliferation and metastasis ability of different groups were detected by CCK-8,plate monoclonal assay and Transwell cell migration assay.The expression levels of ASF1B in GC tissues with positive and negative Helicobacter pylori in gene expression omnibus database(GSE27411)were analyzed,and then the expression levels of ASF1B in GC cell line AGS before and after Helicobacter pylori infection were detected by RT-qPCR and western blot.Result The mRNA and protein expression levels of ASF1B in GC tissues of the database and our cohort were higher than those in adjacent normal tissues.Moreover,higher expression of ASF1B in GC tissues was correlated with the poorer prognosis of GC patients.Compared with siNC group,the ability of cell proliferation and metastasis were reduced in the ASF1B knockdown group(siASF1B#1 group,siASF1B#2 group and siASF1B#3 group).Finally,the expression levels of ASF1B in Helicobacter pylori-positive GC tissues and Helicobacter pylori-infected human GC cells AGS were up-regulated.Conclusion ASF1B may play an important role in the occurrence and development of GC induced by Helicobacter pylori infection.Besides,high expression of ASF1B is associated with earlier metastasis and poorer prognosis of GC patients,which can be used as a molecular marker to predict recurrence and prognosis of GC patients.In addition,knocking down ASF1B significantly inhibited the proliferation and metastasis ability of AGS cell.
关键词
胃癌/抗沉默功能蛋白1B/幽门螺杆菌/细胞增殖/细胞转移
Key words
Gastric cancer/Anti-silencing functional protein 1B/Helicobacter pylori/Cell proliferation/Cell metastasis
维普
万方数据