miR-199调控CFL2蛋白表达对慢性粒细胞白血病细胞增殖和侵袭影响  

Effects of miR-199 on the expression of CFL2 protein in proliferation and invasion of chronic myeloid leukemia cells

作　　者：杨满[1] 吴隼[1] 黄琰[1] 张灵秀[1] 字友梅[1] 吕国庆[1] YANG Man;WU Sun;HUANG Yan;ZHANG Ling-xiu;ZI Yowmei;LU Guo-qing(Department of Hematology, First Affiliated Hospital of Xinxiang Medical College ,Weihui 453100, P. R. China)

机构地区：[1]新乡医学院第一附属医院血液病一科,河南卫辉453100

出　　处：《中华肿瘤防治杂志》2018年第7期466-470,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：河南省教育厅基金(2006320039)

摘　　要：目的肿瘤细胞内的CFL2参与细胞凋亡转移过程,本研究探讨miR-199调控CFL2蛋白的表达对慢性粒细胞白血病细胞增殖和侵袭的影响。方法qPCR检测不同慢性粒细胞白血病细胞株miR-199和CFL2表达水平;分析miR-199表达和慢性粒细胞白血病临床患者复发率和生存期的相关关系;双荧光素酶实验检测miR-199和CFL2之间的相互关系;平板克隆实验检测miR-199对KG-1细胞增殖和凋亡的影响,Transwell侵袭实验检测miR-199对KG-1细胞侵袭能力的影响。平板克隆实验和Transwell侵袭实验检测过表达CFL2后对KG-1细胞增殖和侵袭能力的逆转作用。结果与其他慢性粒细胞白血病细胞株相比,KG-1细胞中miR-199表达为(1.02±0.11)%,明显较高,且CFL2表达为(0.36±0.05)%,明显较低;双荧光素酶实验显示,miR-199可以调控CFL2蛋白的表达水平;平板克隆实验和凋亡实验显示,miR-199可以促进KG-1细胞的增殖,抑制其凋亡行为;Transwell侵袭实验结果显示,miR-199-mimic组细胞侵袭量为426.5±29.6,较对照组(193.6±18.6)明显升高,差异有统计学意义,P=0.016;而过表达CFL2可以逆转其作用。结论 miR-199可以靶向CFL2蛋白影响慢性粒细胞白血病细胞的增殖和侵袭行为。OBJECTIVE CFL2 in tumor cells is involved in the process of apoptosis. The aim of this study was to investi- gate the effect of miR 199 on the proliferation and invasion of human chronic myeloid leukemia cells. METHODS The expression of miR-199 and CFL2 in different chronic myeloid leukemia cell lines was detected by qPCR. The relationships between the expression of miR-199 and the recurrence rate and survival of clinical patients with chronic myeloid leukemia were ana- lyzed. Double luciferase assay was used to detect the relationship between miR-199 and CFL2. The effects of miR-199 on proliferation and apoptosis of KC-I cells were detected by plate cloning assay. Transwell invasion assay was used to detect the effect of miR-199 on the invasion of KG-1 cells. Platelet cloning assay and Transwell invasion assay were used to detect the reversal effect of CFL2 on the proliferation and invasion of KG-1 cells. RESULTS Compared with other chronic mye- loid leukemia cell lines,the expression of miR-199 was （1.02±0.11）% ,which was significantly higher in KG-1 cells,and the expression of CFL2 was （0.36±0.05）%, which was significantly lower. Double luciferase experiments showed that miR-199 could regulate the expression of CFL2 protein,Platelet cloning and apoptosis experiments showed that miR-199 could promote the proliferation of KG-1 cells and inhibit the apoptotic behavior. The results of Transwell invasion assay showed that the cell invasion in the miR-199-mimic group （426.5±29.6） was significantly higher than that in the control group （193.6±18. 6）, the difference was statistically significant （P = 0. 016）. Overexpression of CFL2 can reverse its role. CONCLUSION miR 199 can target CFL2 protein to influence the proliferation and invasion of chronic myeloid leu-kemia cells.

关 键 词：白血病 慢性粒细胞 miR-199 CFL2 侵袭 

分 类 号：R733.72[医药卫生—肿瘤]