通过下调miR-182的表达，CAT104沉默在人白血病细胞中发挥肿瘤抑制作用。
CAT104 silence behaves as a tumor suppressor in human leukemia cells by down regulating miR-182 expression.
作者信息
Zhang Chengfang, Song Guanli, Song Guanbo, Li Ruolei, Gao Min, Zhang Haiguo
机构信息
Department of Clinical Laboratory, Jining No. 1 People's Hospital Jining 272011, Shandong, China.
Department of Preventive and Health Care, Guang'anmen Hospital, China Academy of Chinese Medical Sciences Beijing 100053, China.
出版信息
Int J Clin Exp Pathol. 2017 Dec 1;10(12):11393-11404. eCollection 2017.
BACKGROUND
LncRNAs and miRNAs are found to play crucial roles in the tumorigenesis of acute myeloid leukemia (AML). We aimed to investigate the functions and mechanisms of lncRNA-CAT104 and miR-182 in AML.
METHODS
Expression of CAT104, miR-182, and ZEB1 in K562 and HL60 cell lines was respectively or synchronously altered by transfection. Expressions of CAT104, miR-182 and ZEB1 in cell were then analyzed by qRT-PCR. Cell viability, migration, invasion and apoptosis were evaluated by MTT, transwell assays and flow cytometry, respectively. Protein expressions of ZEB1 and factors related with apoptosis and two signal pathways (Wnt/β-catenin and JNK) were detected by western blot.
RESULTS
CAT104 expressed highly in K562 and HL60 cells compared to embryonic kidney cell line HEK293 (P < 0.001). Knockdown of CAT104 inhibited cell viability, migration and invasion, but increased cell apoptosis of K562 and HL60 cells through inhibitionof miR-182 (P < 0.05). miR-182 promoted cell survival, migration and invasion through upregulatingthe expression of ZEB1 (P < 0.05). miR-182 silence deactivated Wnt/β-catenin and JNK signal pathways by downregulating the expression of ZEB1 in K562 and HL60 cells.
CONCLUSION
LncRNA-CAT104 expressed highly in leukemia cells and its silence inhibited cell survival, migration and invasion by downregulating miR-182 expression. miR-182 functioned as an oncogene by upregulating ZEB1 via which miR-182 silence deactivated Wnt/β-catenin and JNK signal pathways in leukemia cells.
背景
长链非编码RNA（lncRNAs）和微小RNA（miRNAs）在急性髓系白血病（AML）的肿瘤发生中发挥关键作用。我们旨在研究lncRNA-CAT104和miR-182在AML中的功能及机制。
方法
通过转染分别或同步改变K562和HL60细胞系中CAT104、miR-182和ZEB1的表达。然后采用qRT-PCR分析细胞中CAT104、miR-182和ZEB1的表达。分别通过MTT法、Transwell实验和流式细胞术评估细胞活力、迁移、侵袭和凋亡情况。采用蛋白质印迹法检测ZEB1及与凋亡相关的因子以及两条信号通路（Wnt/β-连环蛋白和JNK）的蛋白质表达。
结果
与胚胎肾细胞系HEK293相比，CAT104在K562和HL60细胞中高表达（P<0.001）。敲低CAT104可抑制K562和HL60细胞的活力、迁移和侵袭，但通过抑制miR-182增加细胞凋亡（P<0.05）。miR-182通过上调ZEB1的表达促进细胞存活、迁移和侵袭（P<0.05）。在K562和HL60细胞中，miR-182沉默通过下调ZEB1的表达使Wnt/β-连环蛋白和JNK信号通路失活。
结论
lncRNA-CAT104在白血病细胞中高表达，其沉默通过下调miR-182的表达抑制细胞存活、迁移和侵袭。miR-182通过上调ZEB1发挥癌基因作用，miR-182沉默使白血病细胞中的Wnt/β-连环蛋白和JNK信号通路失活。
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