Glia maturation factor-gamma inhibits the proliferation of human colorectal cancer LoVo cells and affects the cytoskeletal motion of human umbilical vein endothelial cells / 中国组织工程研究

ABSTRACT

BACKGROUND: Glial maturation factor-gamma (GMFG) is one of the members of the ADF/cofilin superfamily protein in the process of cytoskeleton remodeling. It can regulate the cell movement by regulating actin-mediated cytoskeleton reorganization. The authors previously found that the high expression of GMFG is associated with the clinicopathological features of colorectal cancer patients, but the specific upstream and downstream mechanisms need to be explored. OBJECTIVE: To study the effects of GMFG on cytoskeleton and LoVo cell proliferation. METHODS: (1) Human umbilical vein endothelial cells were inoculated in a laser confocal dish. After the cells grew stably, the correlation between the expression of GMFG and the expression of cytoskeletal protein F-actin was observed by immunofluorescence staining. Colchicine inhibited mitosis. The cells were divided into control group, 0.5 mg/L colchicine group and 1.0 mg/L colchicine group. The cell cycle was observed by flow cytometry and GMFG protein expression was observed by western blot assay. (2) Human colorectal cancer LoVo cells were inoculated in a laser confocal dish. After the cells grew stably, the correlation between GMFG expression and cell cycle was observed by immunofluorescence staining. Afterwards, siRNA was used to interfere the expression of GMFG in LoVo cells. The LoVo cells were divided into blank control group, empty transfer group and siRNA-GMFG group. The cell proliferation rate was detected by Edu assay. RESULTS AND CONCLUSION: (1) In human umbilical vein endothelial cells, the expression of GMFG was associated with the cytoskeletal motion of human umbilical vein endothelial cells, and the expression of GMFG increased when the cytoskeleton retracted. After colchicine inhibited the mitosis of human umbilical vein endothelial cells, the proportion of cells in G2/M phase increased and the expression of GMFG protein decreased with the increase of colchicine dosage. (2) In LoVo cells, the expression intensity of GMFG was associated with mitosis, and the expression of GMFG was enhanced in the middle and late stages of cell mitosis. (3) After siRNA interference with GMFG in LoVo cells, the cell proliferation rate was decreased. (4) Results verify that interference with GMFG can inhibit the proliferation rate of LoVo cells of human colorectal cancer, and there is a certain correlation between cytoskeletal motion and GMFG expression.