miR-152对SHI-1细胞增殖、转移及致瘤性的调控机制  

Regulatorg Mchanism of MiR-152 on Proliferation, Metastasis and Tumorigenicity of SHI-1 Cells

作　　者：武坤[1,2,3] 袁峰 祝艳翠[5] 曾云[6,7] WU Kun;YUAN Feng;ZHU Yan-Cui;ZENG Yun(Department of Laboratorial Medicine,The First Affiliated Hospital of Kunming Medical University;Yunnan Key Laboratory of Laboratorial Medicine;Yunnan Institute of Laboratorial Diagnosis;Department of Medical Imaging,The First Affiliated Hospital of Kunming Medical University;Department of Intensive Care Medicine,The First Affiliated Hospital of Kunming Medical University;Department of Hematology,The First Affiliated Hospital of Kunming Medical University;Yunnan Blood Disease Research Center,Kunming 650032,Yunnan Province,China)

机构地区：[1]昆明医科大学第一附属医院医学检验科 [2]云南省检验医学重点实验室 [3]云南省实验诊断研究所 [4]昆明医科大学第一附属医院医学影像科 [5]昆明医科大学第一附属医院重症医学科 [6]昆明医科大学第一附属医院血液科 [7]云南省血液病研究中心,云南昆明650032

出　　处：《中国实验血液学杂志》2019年第5期1455-1462,共8页Journal of Experimental Hematology

基　　金：云南省联合专项基金(2017FE468-035);云南省卫生科技计划(2016NS047);云南省卫生科技计划(2018NS0129)

摘　　要：目的:探究miR-152对人急性单核细胞白血病细胞系SHI-1增殖、转移及致瘤性的调控机制。方法:将购买的SHI-1细胞系进行miR-152过表达(miR-152 agomir组)或抑制处理(miR-152 antagomir组)并设置阴性对照组(NC组)。应用CCK-8法检测各组细胞活力,划痕愈合实验检测各组细胞迁移能力,Transwell法检测各组细胞侵袭能力,Western blot检测各组细胞Cyclin D1、Caspase-3、MMP-2、TIMP-2、E-cadherin和N-cadherin的表达,Annexin V-FITC/PI双染流式细胞术检测各组细胞凋亡情况,裸鼠成瘤实验检测各组细胞的致瘤性。结果:与NC组相比,miR-152 agomir组细胞活力显著下降,细胞迁移和侵袭能力均显著下降(P<0.05),而miR-152antagomir组的细胞活力以及细胞迁移和侵袭能力均显著上升(P<0.05)。与NC组相比,miR-152 agomir组细胞Cyclin D1、MMP-2、N-cadherin蛋白表达均显著下调(P<0.05),但Caspase-3、TIMP-2和E-cadherin的表达在上述各组中明显升高;同时,细胞凋亡增强,裸鼠致瘤性降低(P<0.05)。miR-152 antagomir组Cyclin D1、MMP-2和N-cadherin被显著诱导,但该组Caspase-3、TIMP-2和E-cadherin的蛋白表达显著下调;同时,细胞凋亡减少,裸鼠致瘤性增强(P<0.05)。结论:miR-152能抑制SHI-1细胞系增殖、转移及致瘤的能力,同时诱导细胞凋亡,其机制可能与miR-152调控MMP-2以及TIMP-2等侵袭转移相关因子有关联。Objective:To investigate the regulatory mechanism of miR-152 on proliferation,metastasis and tumorigenesis of human acute monocytic leukemia SHI-1 cells.Methods:The purchased SHI-1 cell line was treated with miR-152 over-expression(miR-152 agomir group)or miR-152 inhibition(miR-152 antagomir group),and the negative control(NC)group was set up.The cell proliferation of each group was detected by CCK-8 assay.Scratch healing assay was employed to determine the migration of cells.Transwell assay was used to measure the invasion of cells.The expressions of Cyclin D1,Caspase-3,MMP-2,TIMP-2,E-cadherin and N-cadherin were detected by Western blot.The flow cyronetry with annexin V-FITC/PI double staining was applied to detect the cell apoptosis.The tumorigenesis of cells was examined by tumor formation experiment in nude mice.Results:Compared with the NC group,the cell proliferation,migration and invasion ability in miR-152 agomir group were significantly decreased(P<0.05),while that in miR-152 antagomir were significantly up-regulated(P<0.05).Compared with the NC group,the protein expression of Cyclin D1,MMP-2,N-cadherin were down-regulated in miR-152 agomir group,but the protein expression of Caspase-3,TIMP-2 and E-cadherin were all up-regulated siginificantly.At the same time,the apoptosis were enhanced,but the timorigenicity in nude mice were significantly decreased(all P<0.05).The protein expression of Cyclin D1,MMP-2,N-cadherin in miR-152 antagomir group,showed high levels but Caspase-3,TIMP-2 and E-cadherin protein showed low levels in comparison with NC group.At the same time,the apoptosis was decreased but the timorigenicity in nude mice was significantly enhanced(all P<0.05).Conclusion:miR-152 can inhibit the proliferation,metastasis and tumorigenesis of SHI-1 cell line,at the same time induce cell apoptosis,thus providing a theoretical basis for the treatment of acute lymphoblastic leukemia.

关 键 词：miR-152 SHI-1细胞 细胞增殖 细胞侵袭 细胞致瘤性 

分 类 号：R733.71[医药卫生—肿瘤]