miR-1269a调控HOXD10基因抑制胆管癌细胞侵袭的作用及其机制  

Role and Mechanism of miR-1269a on Targeted Regulation of HOXD10 in Invasion of Cholangiocarcinoma Cells

作　　者：杨海霞[1,2] 米建强[3] 梁军[1,2] 邵秋菊[1,2] 齐宇红[1,2] 王启明[1,2] 常浩 李恩孝[4] YANG Haixia;MI Jianqiang;LIANG Jun;SHAO Qiuju;QI Yuhong;WANG Qiming;CHANG Hao;LI Enxiao(Department of Radiotherapy,Tangdu Hospital,Fourth Military Medical University,Xi’an 710038,China;Cancer Institute of Fourth Military Medical University,Xi'an 710038,China;Department of Pathology,First Affiliated Hospital of He’nan Science and Technology University,Luoyang 471003,China;Department of Medical Oncology,First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710062,China)

机构地区：[1]第四军医大学唐都医院放疗科,西安710038 [2]第四军医大学肿瘤研究所,西安710038 [3]河南科技大学第一附属医院病理科,洛阳471003 [4]西安交通大学第一附属医院肿瘤内科,西安710062

出　　处：《肿瘤防治研究》2018年第11期894-899,共6页Cancer Research on Prevention and Treatment

摘　　要：目的探讨miR-1269a对HOXD10基因的调控作用及对胆管癌细胞侵袭能力的影响。方法预测并筛选出调控HOXD10基因的miR-1269a作为研究对象;转染miR-1269a模拟物(mimic)及抑制剂(inhibitor)至胆管癌细胞系后检测HOXD10mRNA及蛋白的表达变化,并观察miR-1269a对胆管癌细胞侵袭能力的影响。双荧光素酶报告基因实验验证miR-1269a与HOXD10之间的靶向作用关系。结果miR-1269a在胆管癌组织中表达显著高于癌旁组织(P=0.0023);miR-1269a模拟物可显著降低胆管癌细胞中HOXD10 mRNA(Mz-CHA-1:P=0.0025; RBE:P=0.0038)及蛋白表达水平,且细胞的侵袭能力较对照组显著增强(Mz-CHA-1:P=0.004; RBE:P=0.004)。miR-1269a抑制剂转染则出现相反的结果(QBC939:P=0.16; HCCC9810:P=0.13)。miR-1269a明显抑制野生型HOXD10-3’UTR的荧光素酶活性,而对突变型质粒转染细胞的荧光素酶活性无影响。结论 miR-1269a靶向负性调控HOXD10基因,在胆管癌细胞侵袭过程中发挥重要作用。Objective To explore the possible mechanism of microRNA-1269a(miR-1269a)on the targeted regulation of HOXD10 in the invasion of human cholangiocarcinoma(CCC)cells.Methods We detected six miRNAs levels in human CCC samples and cells for screening miR-1269a as the research object.miR-1269a mimic and inhibitor were transfected into four CCC cells by Lipofectamine liposome respectively.The expressions of HOXD10 mRNA and protein were detected by real-time quantitative PCR(qPCR)and Western blot.The effect of miR-1269a on the invasion of CCC cells was observed.Double luciferase reporter assay was applied to verify the targeting relationship between miR-1269a and HOXD10.Results miR-1269a expression was significantly upregulated in CCC tissues,compared with adjacent normal tissues(P=0.0023).The mRNA and protein levels of HOXD10 in miR-1269a mimic transfection group were lower than those in control group(Mz-CHA-1:P=0.0025;RBE:P=0.0038).miR-1269a mimic significantly elevated the invasion capacity of CCC cells(Mz-CHA-1:P=0.004;RBE:P=0.004),while miR-1269a inhibitor remarkably inhibited the invasion(QBC939:P=0.16;HCCC9810:P=0.13).Double luciferase reporter gene test showed that miR-1269a could significantly inhibit the luciferase activity of wild-type HOXD10-3’UTR,but had no effect on the luciferase activity of mutant plasmid transfected cells.Conclusion miR-1269a may regulate the invasion of CCC cells by targeting HOXD10,and could be used as an effective target for the molecular therapy of CCC.

关 键 词：胆管细胞癌 miR-1269a HOXD10 侵袭 

分 类 号：R735.8[医药卫生—肿瘤]