Effects of lncRNA GIHCG targeting miR-429 on proliferation,migration,and invasion of esophageal squamous cell carcinoma cells
Objective To investigate the effects of long non-coding RNA(lncRNA)GIHCG targeting microRNA(miR)-429 on the proliferation,migration,and invasion of esophageal squamous cell carcinoma(ESCC)cells.Meth-ods Human ESCC cell lines(EC9706,TE1,and KYSE150 cells)and normal esophageal squamous epithelial cell line(Het-1A cells)were routinely cultured.The expression levels of lncRNA GIHCG and miR-429 in the cells were detected by RT-PCR,and the experimental cells were screened.The experimental cells in the logarithmic growth phase were ran-domly divided into the si-NC group and the si-GIHCG group,which were transfected with lncRNA knockdown plasmid neg-ative control and lncRNA GIHCG knockdown plasmid,respectively.The expression levels of lncRNA GIHCG and miR-429 in cells were detected by RT-PCR.The proliferation ability of the cells[optical density(OD)value]was detected by CCK-8.The migration ability of the cells was detected by cell scratch test.The invasion ability of the cells was detected by Transwell chamber test.The expression levels of cell proliferation phenotype proteins[CyclinD1,proliferating cell nuclear antigen(PCNA)]and metastasis phenotype proteins[matrix metalloproteinase 2(MMP-2),matrix metalloproteinase 9(MMP-9)]in the cells were detected by Western blotting.The experimental cells in the logarithmic growth phase were ran-domly divided into the GIHCG-WT+miR-429 mimics group,the GIHCG-WT+NC-mimics group,the GIHCG-MUT+miR-429 mimics group,and the GIHCG-MUT+NC-mimics group,respectively.The targeted relationship between lncRNA GIHCG and miR-429 was verified by dual luciferase reporter gene assay.Results Compared with Het-1A cells,the rela-tive expression levels of lncRNA GIHCG in EC9706,TE1 and KYSE150 were higher(all P<0.05),and the relative expression levels of miR-429 were lower(all P<0.05).Since the relative expression level of lncRNA GIHCG was the high-est and the relative expression level of miR-429 was the lowest in TE1 cells,TE1 cells were used as the experimental cells.Compared with the si-NC group,the relative expression level of lncRNA GIHCG in the si-GIHCG group was lower,the rel-ative expression level of miR-429 was higher,the OD values were lower,the percentage of cell movement distance was lower,and the relative expression levels of CyclinD1,PCNA,MMP-2 and MMP-9 were lower,and the differences were statistically significant(all P<0.05).Compared with the GIHCG-WT+NC-mimics group,the relative luciferase activity in the GIHCG-WT+miR-429 mimics group was lower(P<0.05);there was no statistically significant difference in the rel-ative luciferase activity between the GIHCG-MUT+miR-429 mimics group and the GIHCG-MUT+NC-mimics group(P>0.05).Conclusions The lncRNA GIHCG is highly expressed and the miR-429 is low expressed in ESCC cells.Knock-down of lncRNA GIHCG inhibits the proliferation,migration and invasion of ESCC cells by targeting miR-429.
万方数据
维普
