The effect and mechanism of esophageal cancer related gene 4 on diffuse large B-cell lymphoma
Objective To investigate the effect of esophageal cancer related gene 4(ECRG4)on the proliferation,apoptosis and cycle of diffuse large B-cell lymphoma(DLBCL).Methods The lentivir-us ECRG4-green fluorescent protein(GFP)was prepared and infected in vitro to DLBCL cell lines WSU DLCL2 and DHL4 which low expressed ECRG4 protein.Western blotting and real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR)were used to detect the transcription and expression level of ECRG4 in WSU DLCL2 and DHL4 cells transfected with ECRC4.Hoechst 33342 staining was used to detect the changes of nuclear shape;Cell proliferation activity was detected by methyl thiazolyl tetrazoli-um(MTT)colorimetric method.Cell cycle and apoptosis were observed by flow cytometry,Paired sample t-test was used for comparison between groups.Results Western blotting and real-time fluorescence quan-titative results showed that the transcription and expression levels of ECRG4 in WSU DLCL2 and DHL4 lymphoma cells infected with GFP-ECRC4 lentivirus were higher than those in the control group(24.82±0.02 vs.1.00±0.52).t=4.51,P＜0.05);(34.54±0.03 vs.1.00±0.06,t=4.99,P＜0.05);The results of Hoechst 33342 staining showed that the nuclear shape changed after ECRG4 high expression.Flow cytometry showed that the proportion of WSU DLCL2 and DHL4 cells with ECRG4 overexpression in S phase was higher than that in the non-transfected group[(50.60±4.30)％vs.(43.87±2.60)％,t=9.50,P＜0.05;(48.33±2.15)％vs.(42.56±3.50)％,t=7.85,P＜0.05];MTT results showed that the proliferation of WSU DLCL2 and DHL4 lymphoma cells after ECRG4 overexpression was lower than that of the control group.24 h after ECRG4 transfection[(32.00±0.06)％vs.(60.00±0.05)％,t=3.15,P＜0.05;(56.00±0.03)％vs.(31.00±0.04)％,t=5.32,P＜0.05];After transfection with ECRG4 for 72 h,the ratio of[(47.00±0.03)％vs.(110.00±0.03)％,t=6.15,P＜0.05;(31.00±0.04)％vs.(100.00±0.01)％,t=9.11,P＜0.05].Conclusion ECRG4 gene can inhibit the proliferation of diffuse large B lymphoma cells by blocking cell cycle and inducing apoptosis,and it is a potential tumor suppressor gene.
Esophageal cancer related gene 4Diffuse large B lymphoma cellsCell proliferationCell apoptosis
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