Research Paper Volume 13, Issue 3 pp 3866—3885

Long intergenic non-coding RNA LINC00485 exerts tumor-suppressive activity by regulating miR-581/EDEM1 axis in colorectal cancer

LINC00485 modulates CRC cell proliferation, migration, and invasion by regulating miR-581/EDEM1 axis. (A) The transfection efficiency of small interfering RNA targeting EDEM1 was validated by RT-qPCR analysis and western blotting assay. (B) miR-581 levels are significantly down-regulated both in LINC00485-overexpressed LoVo cells and LINC00485-overexpressed LoVo cells transfected with siEDEM1. (C) EDEM1 expression increases in LINC00485-overexpressed LoVo cells, but decreases in LINC00485-overexpressed LoVo cells after transfection with siEDEM1. (D) Cell viability of LINC00485-overexpressed LoVo cells with or without siEDEM1 treatment was measured by the CCK-8 assy. (E) The colony formation assay results showing that overexpression of LINC00485 significantly inhibits the colony forming ability of LoVo cell, but EDEM1 knockdown reverses the result induced by LINC00485 overexpression. (F) Transwell migration and (G) invasion assays showing the effect of EDEM1 knockdown on LINC00485-overexpressed LoVo cells. (H) The mRNA levels of EDEM1, miR-581, cytokeratin, E-cadherin, N-cadherin, and vimentin in LINC00485-overexpressed LoVo cells transfected with or without siEDEM1. Comparison between two groups were assessed using student’s t-test. Multiple comparison was analyzed using the one-way ANOVA with LSD test. Bars were represented as S.D. *PPP

Figure 6. LINC00485 modulates CRC cell proliferation, migration, and invasion by regulating miR-581/EDEM1 axis. (A) The transfection efficiency of small interfering RNA targeting EDEM1 was validated by RT-qPCR analysis and western blotting assay. (B) miR-581 levels are significantly down-regulated both in LINC00485-overexpressed LoVo cells and LINC00485-overexpressed LoVo cells transfected with siEDEM1. (C) EDEM1 expression increases in LINC00485-overexpressed LoVo cells, but decreases in LINC00485-overexpressed LoVo cells after transfection with siEDEM1. (D) Cell viability of LINC00485-overexpressed LoVo cells with or without siEDEM1 treatment was measured by the CCK-8 assy. (E) The colony formation assay results showing that overexpression of LINC00485 significantly inhibits the colony forming ability of LoVo cell, but EDEM1 knockdown reverses the result induced by LINC00485 overexpression. (F) Transwell migration and (G) invasion assays showing the effect of EDEM1 knockdown on LINC00485-overexpressed LoVo cells. (H) The mRNA levels of EDEM1, miR-581, cytokeratin, E-cadherin, N-cadherin, and vimentin in LINC00485-overexpressed LoVo cells transfected with or without siEDEM1. Comparison between two groups were assessed using student’s t-test. Multiple comparison was analyzed using the one-way ANOVA with LSD test. Bars were represented as S.D. *P<0.05; **P<0.01; ***P<0.001. NS, not significant; OE, overexpression; si, small interfering RNA targeting EDEM1; EDEM1, ER-degradation-enhancing alpha-mannosidase-like protein-1.