Overexpresssion of NDUFA4 induces epithelial-mesenchymal transition of human colorectal cancer cells / 医学研究生学报

ABSTRACT

Objective The role and action mechanisms of NADH dehydrogenase 1 alpha subcomplex 4 (NDUFA4) in the development of human colorectal cancer (CRC) are not yet clarified. This article aims to study the effect of overexpressed NDUFA4 on the epithelial-mesenchymal transition (EMT) of human CRC cells. Methods The recombinant plasmid NDUFA4 (p-NDUFA4) and control plasmid (p-Cont) were transiently transfected into human CRC HCT116 cells. The expression of NDUFA4 in the cells was determined by real-time PCR and Western blot respectively, and the migration ability of the cells detected by Transwell migration and wound healing assays. The expression levels of MMP2, MMP9 and CXCR4 in the cells were measured by qRT-PCR, and those of Twist, Snail, E-cadherin and Vimentin by Western blot and by immunofluorescence assay, respectively. Results Compared with the p-Cont group, the human CRC HCT116 cells of the p-NDUFA4 group showed significantly up-regulated expressions of NDUFA4 mRNA ([0.96±0.15]% vs [1.94±0.08]%, P<0.05) and NDUFA4 protein ([0.06±0.05]% vs [1.07±0.12]%, P<0.05), increased in vitro migration ability ([29.51±3.17]% vs [54.36±4.08]%, P<0.01) and migrated cell rate ([0.99±0.12]% vs [1.85±0.10]%, P<0.01), elevated expression levels of MMP2, MMP9, CXCR4, N-cadherin,Vimentin, Snail and Twist, but down-regulated level of E-cadherin (P<0.05). Conclusion Overexpressed NDUFA4 promotes the epithelial-mesenchymal transition of human colorectal cancer cells.