Objective This paper is to investigate the effect of miR-15b-3p targeting CKLF-like MARVEL transmembrane domain-containing protein 6(CMTM6)on epithelial-mesenchymal transformation(EMT)in gastric cancer cells.Methods The expression levels of miR-15b-3p and CMTM6 in human gastric cancer cells SGC-7901,BGC-823,MKN-1,and normal gastric mucosal epithelial cells GES-1 were detected by real-time fluorescence quantitative PCR.The dual luciferase reporter gene assay was used to detect the targeting relationship between miR-15b-3p and CMTM6.BGC-823 cells were randomly divided and assingned to the control group(no treatment),the miR-15b-3p inhibitor group(transfected with miR-15b-3p inhibitor),and the miR-15b-3p inhibitor+shRNA-CMTM6 group(transfected with miR-15b-3p inhibitor and shRNA-CMTM6).The MTT assay,scratch healing assay,and Transwell assay were used to detect cell proliferation,migration,and invasion.The expression level of EMT-related proteins was detected by Western blotting.Results Compared with GES-1 cells,the relative expression levels of miR-15b-3p and CMTM6 mRNA in SGC-7901,BGC-823,and MKN-1 cells are significantly increased(P＜0.05).The results of double luciferase reporter gene assay show that compared with the miR-NC+CMTM6-WT group,the relative luciferase activity of the miR-15b-3p inhibitor+CMTM6-WT group is significantly decreased(P＜0.05).Compared with the control group,the relative expression levels of miR-15b-3p and CMTM6 mRNA in the miR-15b-3p inhibitor group are significantly decreased(P＜0.05).Compared with the miR-15b-3p inhibitor group,the relative expression level of miR-15b-3p in the miR-15b-3p inhibitor+shRNA-CMTM6 group has no significant changes(P＞0.05),but the relative expression level of CMTM6 mRNA is significantly decreased(P＜0.05).Compared with the control group,the cell proliferation inhibition rate in the miR-15b-3p inhibitor group is increased;the cell scratch healing rate,the number of cell invasion,and the protein expression levels of N-cadherin and Vimentin are decreased,but the protein expression level of E-cadherin is increased,with statistically significant differences(P＜0.05).Compared with the miR-15b-3p inhibitor group,the cell proliferation inhibition rate in the miR-15b-3p inhibitor+shRNA-CMTM6 group is increased;the cell scratch healing rate,the number of cell invasion,and the protein expression levels of N-cadherin and Vimentin are decreased,but the protein expression level of E-cadherin is increased,with statistically significant differences(P＜0.05).Conclusion Down-regulation of miR-15b-3p expression can inhibit the proliferation,migration,invasion ability and EMT level of gastric cancer cells by targeting CMTM6.
Gastric cancermiR-15b-3pCKLF-like MARVEL transmembrane domain-containing protein 6Epithelial-mesenchymal transitionProliferationInvasionMigrate
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