Effects of epidermal growth factor-like domain-3 on proliferation,apoptosis,migration,and inva-sion of SNU-449 hepatocellular carcinoma cells
Objective To investigate the influence of epidermal growth factor-like domain-3(EGFL3)on the proliferation,apoptosis,migration,and invasion of SNU-449 human hepatocellular carci-noma cells,while also delving into the underlying molecular mechanisms.Methods Lentivirus-mediated small interfering RNA technology was used to construct EGFL3 knockdown SNU-449 cell lines,and real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR)technology was used to detect the interference efficiency of the EGFL3 gene,and the group with the highest knockdown efficiency was se-lected as the knockdown group(KD group),and the negative control group(NC group)was set up,and then the methyl thiazolyl tetrazolium(MTT)assay,cell clone formation assay,apoptosis assay,Transwell assay and Transwell invasion assay were performed to compare the proliferation,apoptosis,invasion and metastasis of the two SNU-449 cell lines,and finally,the relative expression of transforming growth factor-β1(TGF-β1),SMAD gene relative expression levels were detected.Independent sample t-test was used for comparison of samples between groups.Results Compared with the NC group,the knockdown efficiency of EGFL3 gene in the KD group was 75.7％(t=13.730,P＜0.01),and the growth rate of SNU-449 cells in the KD group was significantly slowed down(5.29±0.23 vs.6.30±0.26,t=6.606,P＜0.01;1±1 vs.19±2,t=14.690,P＜0.01),apoptosis rate increased significantly[(11.06± 0.29)％vs.(3.20±0.13)％,t=-43.280,P＜0.01],and the number of metastasized and infiltrated cells decreased significantly[(104±6)cells vs.(178±3)cells,t=20.090,P＜0.01;(209.00± 0.00)cells vs.(306.00±3.51)cells,t=48.170,P＜0.01].The RT-qPCR assay showed that the rela-tive expression of TGF-β1,SMAD2,SMAD3 and SMAD4 genes was significantly up-regulated in the SNU-449 cell line of the KD group as compared with that of the NC group(t=3.759,5.464,5.979,8.956,and P＜0.05),while the relative expression of SMAD7 gene was slightly down-regulated(t=0.368,P＞0.05).Conclusion EGFL3 gene promotes growth,proliferation,invasion,metastasis and in-hibits apoptosis of SNU-449 hepatocellular carcinoma cells by a mechanism that may be related to the inhi-bition of TGF-β1/SMAD signaling pathway.
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