Family with sequence similarity 107 member A suppresses prostate cancer progression:an experi-mental study
Objective To investigate the effects of family with sequence similarity 107 member A(FAM107A)on the migration,invasion and apoptosis of prostate cancer(PCa)by regulating hypoxia in-ducible factor 3 subunit alpha(HIF3A).Methods The target genes were screened and analyzed using bioinformatics tools.Human prostate cancer cells PC-3 were cultured in vitro,then stably transfected with retrovirus and divided into separate groups.The expression levels of HIF3A and DNA methyltransferase 1(DNMT1)were confirmed through Western blotting.Flow cytometry was employed to detect cell cycle and apoptosis.In addition,pyrosequencing was employed to evaluate promoter methylation levels.Cell func-tional assays were performed to investigate the effects on migration and invasion of PCa cells.Differential expression between two groups was compared using independent samples t-test.Results Bioinformatics a-nalysis revealed that the expression of FAM107A gene was lower in PCa tissues than in normal prostate tis-sues(P＜0.01),and its expression level decreased with higher Gleason score(GS)(P＜0.01).Scratch test and Transwell invasion assay demonstrated that overexpression of FAM107A inhibited the migration(C4-2:0.337±8.930 vs.29.334±2.651,t=-7.260,P＜0.05;PC-3:18.672±10.494 vs.52.043±3.512,t=-6.024,P＜0.05)and invasion(DU145:117.703±12.845 vs.242.303±21.401,t=10.273,P＜0.05;PC-3:51.674±9.502 vs.139.303±13.394,t=16.025,P＜0.05)of PCa cells,while promoting apoptosis(17.180±0.700 vs.0.527±0.121,t=40.600,P＜0.05).Cell cycle experiment results indicate that the proportion of cells in the G1 phase in the oe-FAM107A-ENZ group was higher than in the control group(80.410±0.786 vs.35.583±0.714,t=73.130,P＜0.05),while the proportion of cells in the S phase was lower than in the control group(11.233±0.374 vs.38.747±1.809,t=25.790,P＜0.05),and cell division was arrested at the S phase.Western blotting analysis demonstrated that the expression level of DNMT1 in the overexpressed FAM107A group was lower than in the control group(0.277±0.032 vs.0.900±0.026,t=25.93,P＜0.05),and the expression level of HIF3A was higher than in the control group(0.657±0.040 vs.0.523±0.043,t=4.041,P＜0.05).The expression level of HIF3A was significantly restored in the sh-FAM107A-5-aza group(0.550±0.020 vs.0.42±0.067,t=3.073,P＜0.05).Western blotting analysis demonstrated that the expres-sion level of E-cadherin was higher than in the control group when FAM107A was overexpressed(0.773±0.095 vs.0.307±0.046,t=7.683,P＜0.05),while the expression level of N-cadherin was lower than in the control group(0.380±0.060 vs.0.733±0.045,t=8.154,P＜0.05),and the expression level of Vimentin was also lower than in the control group(0.347±0.107 vs.0.700±0.040,t=5.361,P＜0.05).Conclusion High expression of FAM107A can inhibit the methylation of the HIF3A promoter,promoting its expression,thereby inhibiting the migration and invasion of PCa.This may serve as a poten-tial biomarker for the progression of PCa.
Prostate cancerFamily with sequence similarity 107 member AHypoxia inducible factor 3AMethyltransferase 1Cell function
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