神经胶质成熟因子γ抑制人结直肠癌LoVo细胞增殖并影响人脐静脉内皮细胞的骨架运动  

Glia maturation factor-gamma inhibits the proliferation of human colorectal cancer LoVo cells and affects the cytoskeletal motion of human umbilical vein endothelial cells

作　　者：王惠丽[1] 陈志江[2] 吴炳义[3] Wang Huili;Chen Zhijiang;Wu Bingyi(Clinical Laboratory,Guangdong Women and Children Hospital,Guangzhou 511442,Guangdong Province,China;Pediatric Center,Zhujiang Hospital of Southern Medical University,Guangzhou 510280,Guangdong Province,China;Clinical Medicine Research Center,Nanfang Hospital,Southern Medical University,Guangzhou 510515,Guangdong Province,China)

机构地区：[1]广东省妇幼保健院检验科,广东省广州市511442 [2]南方医科大学珠江医院儿科中心,广东省广州市510280 [3]南方医科大学南方医院临床医学研究中心,广东省广州市510515

出　　处：《中国组织工程研究》2021年第13期2055-2059,共5页Chinese Journal of Tissue Engineering Research

基　　金：2020年广东省医学科学技术研究基金项目(A2020599),项目负责人:王惠丽。

摘　　要：背景:神经胶质成熟因子γ(glia maturation factor-gamma,GMFG)是细胞骨架运动重建过程中ADF/cofilin超家族蛋白成员之一,可以通过调节肌动蛋白介导细胞骨架重组参与细胞运动的调控。作者前期发现GMFG的高表达与结直肠癌肿瘤患者临床病理相关,但具体的上游及下游机制有待探究。目的:探索GMFG对人脐静脉内皮细胞骨架及LoVo细胞增殖的影响。方法:①将人脐静脉内皮细胞接种在激光共聚焦皿中,待细胞贴壁稳定生长后,细胞免疫荧光染色观察GMFG的表达与人脐静脉内皮细胞骨架蛋白F-actin表达的关系;然后加入秋水仙素抑制细胞有丝分裂,分为对照组、0.5 mg/L秋水仙素组、1.0 mg/L秋水仙素组,流式细胞术观察3组细胞周期的变化,Western blot蛋白印迹观察3组细胞GMFG蛋白表达;②将人结直肠癌LoVo细胞接种在激光共聚焦皿中,待细胞贴壁稳定生长后,细胞免疫荧光染色观察GMFG的表达与细胞周期的相关性,然后采用siRNA技术干扰LoVo细胞中GMFG的表达,分为空白对照组、空转染组和siRNA-GMFG组,Edu实验检测细胞增殖率的变化。结果与结论:①在人脐静脉内皮细胞中,GMFG的表达与人脐静脉内皮细胞骨架运动有关,细胞骨架回缩时GMFG表达增多;秋水仙素抑制人脐静脉内皮细胞有丝分裂后,随秋水仙素用量增加G2/M期细胞占比增加,GMFG蛋白表达下降;②在LoVo细胞中,GMFG的表达强度与细胞有丝分裂有关,在细胞有丝分裂中后期GMFG表达增强;③siRNA技术干扰LoVo细胞GMFG表达后,细胞增殖率下降;④结果表明,干扰GMFG表达能够抑制人结直肠癌LoVo细胞的增殖率,并且细胞骨架运动与GMFG的表达存在一定的相关性。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 t

关 键 词：人脐静脉内皮细胞 细胞骨架 LOVO细胞 细胞增殖 神经胶质成熟因子γ 秋水仙素 肠癌 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]