A study on the mechanism of SNHG3 promoting proliferation,migration,invasion and epithelial-mesenchymal transition of hepatoma cells by regulating the miR-186-5p/ZEB1 axis

CHEN Xiaolan; SU Yayong; LIU Shuangping; WANG Lihui; XU Chengrun

doi:10.3969/j.issn.1674-3806.2024.04.12

Abstract：Objective To explore the mechanism of SNHG3 promoting proliferation,migration,invasion and epithelial-mesenchymal transition(EMT)of hepatoma cells by regulating the micro ribonucleic acid(miR)-186-5p/zinc finger E-box binding homeobox 1(ZEB1)axis.Methods Human-derived hepatoma(HepG2)cells were transfected with small interfering RNA(siRNA)to knock out SNHG3 and transfected with pcDNA3.1(+)/SNHG3 to overexpress SNHG3,and real-time fluorescent quantitative polymerase chain reaction(FQ-PCR)was used to detect whether the trans-fections were successful.HepG2 cell proliferation was detected by colony formation assay.Cell scratch wound healing assay was used to detect the migration rate of HepG2 cells.Transwell assay was used to detect the number of HepG2 cell invasion.Dual-luciferase reporter assay was used to verify the targeting relationship between SNHG3 and miR-186-5p as well as the targeting relationship between miR-186-5p and ZEB1.The expression levels of SNHG3,ZEB1,E-cadherin,N-cadherin,matrix metalloproteinase-2(MMP-2),vimentin,matrix metalloproteinase-9(MMP-9)and Snail were detected by using FQ-PCR and Western blot.Results The expression of SNHG3 in hepatocellular carcinoma(HCC)tissues was higher than that in normal tissues,and the survival prognosis of the HCC patients with low/moderate SNHG3 expression was better than that of the HCC patients with high SNHG3 expression,and the differences were statistically significant(P＜0.05).The results of cloncolony formation assay showed that compared with that in the pcDNA3.1(+)group,the pro-liferation of HepG2 cells in the pcDNA3.1(+)/SNHG3 group was significantly increased(P＜0.05).The results of cell scratch wound healing assay showed that the migration rate of HepG2 cells in the pcDNA3.1(+)/SNHG3 group was significantly higher than that in the pcDNA3.1(+)group(P＜0.05).The results of Transwell assay showed that the number of HepG2 cell invasion in the pcDNA3.1(+)/SNHG3 group was significantly higher than that in the pcDNA3.1(+)group(P＜0.05).The results of dual-luciferase reporter assay showed that after co-transfection with miR-186-5p mimic and pGL6-SNHG3-WT,the luciferase activity was lower than the other groups,and after co-transfection with miR-186-5p mimic and pGL6-ZEB1-WT,the luciferase activity was lower than the other groups,and the differences were statistically significant(P＜0.05).The results of FQ-PCR and Western blot showed that when SNHG3 was overexpressed,the relative expressions of E-cadherin mRNA and protein were significantly downregulated,while the relative expressions of N-cadherin,MMP-2,vimentin,MMP-9,and Snail mRNA and protein,and the relative expressions of ZEB1 protein were signifi-cantly upregulated,and after SNHG3 was knocked out,the relative expressions of E-cadherin mRNA and protein were significantly upregulated,while the relative expressions of N-cadherin,MMP-2,vimentin,MMP-9 and Snail mRNA and protein,and the relative expression of ZEB1 protein were significantly downregulated,and the differences were statistically significant(P＜0.05).Conclusion SNHG3 can promote the proliferation,migration,invasion and EMT of HCC cells by regulating the miR-186-5p/ZEB1,indicating that SNHG3 is a potential therapeutic target for HCC.