Effects of different transcriptional variants of platelet-derived growth factor A on proliferation and invasion of bladder cancer cells
Objective To explore the effects of different transcriptional variants of platelet-derived growth factor A(PDGFA),PDGFA-L and PDGFA-S,on the proliferation and migration of bladder cancer cells.Methods The exome sequencing data in the cancer genome atlas(TCGA)were analyzed by bioin-formatics methods to screen out abnormal alternative splicing events of PDGFA in bladder cancer.PDGFA-L and PDGFA-S were overexpressed in bladder cancer cell lines(EJ and T24).The experimental groups included EJ and T24 cell lines control group,EJ(EJ-PDGFA-L group and EJ-PDGFA-S group)and T24 cell lines(T24-PDGFA-L group and T24-PDGFA-S group)stably overexpressing PDGFA.Cell count-ing kit-8(CCK-8)assay,plate clone formation assay and Transwell assay were used to observe the effect of PDGFA on the proliferation and migration of bladder cancer.Independent sample t test was used for com-parison between groups.Results Through bioinformatics analysis of the exon sequencing data of PDGFA gene in each tumor,abnormal alternative splicing of PDGFA was found in a variety of cancers,including bladder cancer.As compared with normal tissues,the expression of PDGFA-L in tumor tissues was higher than that of PDGFA-S(P＜0.05).The results of CCK-8 showed that as compared with the control group,the proliferation ability of PDGFA overexpression group was significantly increased[the optical density of EJ cells in PDGFA-L and PDGFA-S groups was significantly higher than that in the control group[(1.958±0.081 vs.0.982±0.357,t=12.765,P＜0.05)and(1.324±0.476 vs.0.982±0.357,t=17.977,P＜0.05)].In T24 cell lines,the optical density of PDGFA-L and PDGFA-S groups was sig-nificantly higher than that of the control group[(1.619±0.053 vs.0.821±0.029,t=17.61,P＜0.05)and(1.219±0.073 vs.0.821±0.029,t=16.32,P＜0.05)].The results of colony formation assay showed that the proliferation ability of PDGFA overexpression group was significantly higher than that of control group[the colony number of EJ cell line in PDGFA-L and PDGFA-S group colony number was sig-nificantly greater than that in the control group(235.50±10.50 vs.160.10±2.11,t=5.12;P＜0.05)and(181.30±6.32 vs.160.10±2.11,t=3.15,P＜0.05).The colony number of T24 cell line in the PDGFA-L and PDGFA-S groups was significantly greater than that in the control group[(590.0±27.0 vs.351.0±16.0,t=6.713,P＜0.05)and(527.0±23.3 vs.351.0±16.0,t=6.907,P＜0.05)].Transwell assay showed that the migration ability of PDGFA overexpression group was significantly higher than that of control group[the number of migrating EJ cells in the PDGFA-L and PDGFA-S groups was sig-nificantly greater than that in the control group[(635.0±15.0 vs.125.0±7.5,t=17.32,P＜0.05)and(611.0±19.0 vs.125.0±7.5,t=18.55,P＜0.05)].The number of migrating T24 cells in the PDGFA-L and PDGFA-S groups was significantly greater than that in the control group[(733±13 vs.117±9,t=21.18,P＜0.05)and(607±12 vs.117±9,t=22.37,P＜0.05)].Conclusion PDGFA can promote the proliferation and invasion of bladder cancer cells,and the full-length transcript PDGFA-L has a stronger cancer-promoting effect.
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