长链非编码RNA NKILA调节miR-889-3p介导肝细胞癌进展的研究  

Long non-coding RNA NKILA mediates the progression of hepatocellular carcinoma through regulating miR-889-3p

作　　者：王红梅[1] 陈思瑞[1] 杜红[1] 曹佳 Wang Hongmei;Chen Sirui;Du Hong;Cao Jia(Department of Hepatobiliary and Breast Surgery,Mianyang Central Hospital,Mianyang 621000,China;Department of Hepatobiliary and Breast Surgery,Affiliated Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610000,China)

机构地区：[1]四川省绵阳市中心医院肝胆乳腺外科,绵阳621000 [2]成都中医药大学附属医院肝胆乳腺外科,成都610000

出　　处：《广西医科大学学报》2021年第4期627-635,共9页Journal of Guangxi Medical University

基　　金：supported by a grant from the Key projects of science and Technology Department of Sichuan Province(No. 2019SZ0069)。

摘　　要：目的:探究NKILA在肝癌发展过程中的潜在作用。方法:采用实时荧光定量PCR(qRT-PCR)法检测HCC组织和癌旁组织、肝癌细胞HepG2和正常肝细胞L-02中NKILA和miR-889-3p的相对表达量。分别通过克隆形成和血管生成实验来评估细胞生长和肿瘤血管生成。免疫蛋白印迹用于检测上皮间充质转化(EMT)相关蛋白的表达。NKILA与miR-889-3p之间的联系采用荧光素酶报告基因分析。建立小鼠移植瘤模型,免疫组化检测其相关蛋白的表达。结果:miR-889-3p是NKILA的直接靶标,NKILA负向调控miR-889-3p的表达。q RT-PCR分析显示,在肝癌组织和HepG2细胞中NKILA表达明显下调,而miR-889-3p表达上调。NKILA过表达通过下调miR-889-3p而抑制HCC细胞增殖、EMT和肿瘤血管生成。体内模型显示NKILA显著降低异种移植肿瘤的体积和重量,并伴有E-钙粘蛋白的增加和VEGF的减少。此外,免疫组化实验发现单独使用NKILA可以减少血管内皮细胞标记物(CD34和CD105)的阳性表达。结论:过表达的NKILA通过下调miR-889-3p抑制HCC的发生,miR-889-3p可能是HCC患者的新治疗靶点。Objective:To explore the potential role of NKILA in the process of hepatocellular carcinoma(HCC) development. Methods:The relative expression of NKILA and mi R-889-3 p in HCC tissues and cells was quantified by quantitative real-time reverse transcription PCR(q RT-PCR). Cell growth and tumor angiogenesis were assessed by colony formation and angiogenesis assay, respectively. Western blotting was used to detect epithelial-mesenchymal transition(EMT)-related proteins. The association between NKILA and mi R-889-3 p was conducted by luciferase reporter analysis. A model of mouse xenograft tumor was established to detect the expression of related proteins by immunohistochemistry(IHC) analysis.Results: mi R-889-3 p was a direct target of NKILA, and NKILA negatively regulated mi R-889-3 p. q RT-PCR assay showed the NKILA expression was conspicuously down-regulated and mi R-889-3 p expression was up-regulated in HCC tissues and cells. NKILA overexpression suppressed HCC cell proliferation, EMT and tumor angiogenesis through directly sponging mi R-889-3 p. More importantly, in vivo model displayed NKILA significantly reduced the volume and weight of xenograft tumor, accompanied by E-cadherin increase and VEGF decrease. Furthermore, IHC revealed NKILA alone reduced the number of vascular endothelial cell markers(CD34 and CD105). Conclusion: NKILA overexpression curbed the development of HCC via regulating mi R-889-3 p, which may be a novel therapeutic target in patients with HCC.

关 键 词：长的非编码RNA 恶性肿瘤 上皮-间质转化 肿瘤血管生成 

分 类 号：R735.7[医药卫生—肿瘤]