Effect of ZNF750 on the biological behavior of endometrial cancer cells and its mechanism
Objective To study the role of zinc finger protein 750(ZNF750)in the proliferation,invasion and migration of endometrial cancer cells and to screen out its potential target genes.Methods Real time fluorescence quantitative poly-merase chain reaction(qRT-PCR)and western blotting were used to detect the mRNA and protein expression levels of ZNF750 in endometrial cancer cell line Ishikawa after transfection of ZNF750 overexpression plasmid(OE-ZNF750 group)and ZNF750 interference fragment(si-ZNF750 group).Cell counting kit 8(CCK8)was used to detect cell proliferation a-bility,while Trans well assay was used to detect cell invasion and migration ability.Gene expression microarray analysis combined with bioinformatics methods to screen candidate target genes regulated by ZNF750.Set up a control group(NC group),and compare between groups by using t-tests and non parametric tests.Results At 72 hours of upregulation of the ZNF750 gene,the CCK8 experiment showed that compared with the NC group(5.79±0.20),the proliferation abili-ty of Ishikawa cells in the OE-ZNF750 group(4.70±0.10)decreased,t=10.571,P＜0.001.Transwell experiment showed that the number of cell invasions and migrations in the NC group was(156.44±7.84)and(125.55±19.69),while the number of cell invasions and migrations in the OE-ZNF750 group was(103.44±19.21)and(49.33±14.15),with statistically significant differences.The t-values were 7.660 and 9.428,respectively,both P＜0.001.After down-regulating the ZNF750 gene for 72 hours,the CCK8 experiment results showed that compared with the NC group(5.21±0.07),the proliferation ability of Ishikawa cells in the si-ZNF750 group(5.59±0.12)was increased,t=-5.876,P＜0.001.Transwell experiment results showed that the invasion number of cells in the NC group was 159.11±32.91,migration number was 84.88±11.47,and the invasion number of cells in the si-ZNF750 group was 314.77±24.06,migration number was 181.11±18.01,and the differences were statistically significant,with t-values of-11.454 and-13.518,respectively,both P＜0.001.Through gene expression microarray analysis,downstream target genes of ZNF750 were screened.The results showed that after upregulating ZNF750,there were 414 differentially expressed genes,and after downregulating ZNF750,there were 50 differentially expressed genes.Combined with bioinformatics,cancer related genes were screened,including RAC2,KLF4,FN1,IGF2,and MAGEA2.The qRT-PCR results showed that after up/down regulation of ZNF750,compared with the NC group,the mRNA expression levels of KLF4,RAC2,MAGEA2 and IGF2 significantly increased/decreased,while the mRNA expression levels of FN1 significantly decreased/increased,with statistical significance(all P＜0.05).Conclusion In endometrial cancer cells,ZNF750 acts as a tumor suppressor gene,inhibiting the proliferation,invasion and migration of endometrial cancer cells.RAC2,KLF4,FN1,IGF2 and MAGEA2 are potential candidate target genes for ZNF750.
zinc-finger protein 750endometrial carcinomaproliferationinvasionmigration
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