LncRNASNHG14靶向miR-449a/CCNE2对前列腺癌细胞生物行为学的影响  

Effects of LncRNA SNHG14 on the biological behaviors of prostate cancer cells by targeting miR-449a/CCNE2

作　　者：刘晶 张国民 李强 王亮 刘志飞[2] Liu Jing;Zhang Guomin;Li Qiang;Wang Liang;Liu Zhifei(Department of Urology,Linxi Hospital,Kailuan General Hospital,Tangshan 063100,Hebei,China;Department of Urology,Tangshan People's Hospital,Tangshan 063100,Hebei,China)

机构地区：[1]开滦总医院林西医院泌尿外科,河北唐山063100 [2]唐山市人民医院泌尿外科,河北唐山063000

出　　处：《中国男科学杂志》2024年第5期49-57,共9页Chinese Journal of Andrology

摘　　要：目的探究长链非编码RNA(LncRNA)核内小RNA宿主基因14(SNHG14)靶向微小RNA-449a(miR-449a)/细胞周期蛋白E2(CCNE2)对前列腺癌(PCa)细胞生物行为学的影响。方法用qRT-PCR法检测前列腺组织及癌细胞中LncRNASNHG14、miR-449a、CCNE2 mRNA水平;用双荧光素酶实验检测LncRNASNHG14与miR-449a及CCNE2与miR-449a的靶向关系;将DU145细胞分为Control组、si-NC、si-SNHG14、si-SNHG14+anti-NC、si-SNHG14+anti-miR-449a,除Control组外,其余各组转染相应质粒;用Edu实验检测DU145细胞增殖;用流式细胞仪检测DU145细胞凋亡;用划痕愈合实验检测DU145细胞迁移;用Transwell实验检测DU145细胞侵袭;用Western blot检测Ki67、Cyclin D1、Bax、caspase-3、CCNE2蛋白表达;用裸小鼠移植瘤检测LncRNA SNHG14对前列腺移植瘤生长以及对miR-449a/CCNE2的影响。结果在PCa组织及细胞中LncRNA SNHG14、CCNE2呈高表达,miR-449a呈低表达(P<0.05);生物信息学、双荧光素酶分析显示LncRNA SNHG14与miR-449a及CCNE2与miR-449a存在靶向结合位点;与si-NC组相比,si-SNHG14组细胞LncRNASNHG14、CCNE2、Edu阳性率、Cyclin D1、Ki67表达水平、划痕愈合率、侵袭细胞数目降低,miR-449a、细胞凋亡率、Bax、caspase-3表达水平升高(P<0.05);与si-SNHG14+anti-NC组相比,si-SNHG14+anti-miR-449a组miR-449a、细胞凋亡率、Bax、caspase-3表达水平降低,CCNE2、Edu阳性率、Cyclin D1、Ki67表达水平、划痕愈合率、侵袭细胞数目升高。裸小鼠移植瘤实验结果显示,si-SNHG14组裸小鼠移植瘤体积、质量比si-NC组更小,LncRNASNHG14、CCNE2表达量下降,miR-449a表达量上升(P<0.05)。结论在PCa细胞中下调LncRNA SNHG14可通过调节miR-449a/CCNE2信号通路,抑制癌细胞增殖、迁移、侵袭,促进癌细胞凋亡。Objective To investigate the effects of long non coding RNA(LncRNA)small nucleolar RNA host gene 14(SNHG14)on the biological behaviors of prostate cancer(PCa)cells by targeting miR-449a/cyclin E2(CCNE2).Methods The expressions of LncRNA SNHG14,miR-449a,and CCNE2 mRNA in prostate cancer tissues and cancer cells were detected by qRT-PCR;The targeting relationship between LncRNA SNHG14 and miR-449a,and between CCNE2 and miR-449a were analyzed by dual luciferase assay;DU145 cells were divided into 5 groups including the control group,si-NC group,si-SNHG14 group,si-SNHG14+anti-NC group,si-SNHG14+anti miR-449a group.Except for the cells in the control group,the cells in all other groups were transfected with corresponding plasmids;Proliferation of DU145 cells was measured by EdU assay;Apoptosis of DU145 cells was analyzed by flow cytometry;Migration of DU145 cells was evaluated by wound healing assay;Invasion of DU145 cells was examined by Transwell invasion assay;The expressions of Ki67,Cyclin D1,Bax,caspase-3,and CCNE2 proteins were detected by western blot;Subcutaneous tumor nude mouse model was used to assess the effects of LncRNA SNHG14 on the growth of tumors and miR-449a/CCNE2.Results LncRNA SNHG14 and CCNE2 were highly expressed in PCa tissues and cells,while miR-449a was lowly expressed(P<0.05);Dual luciferase analyses showed there were targeted binding sites between LncRNA SNHG14 and miR-449a,and between CCNE2 and miR-449a;Compared with the si-NC group,the expression levels of LncRNA SNHG14,CCNE2,Cyclin D1 and Ki67,the positive rate of Edu,wound healing rate,and number of invaded cells in the si-SNHG14 group were all significantly decreased,whereas the expressions of miR-449a,Bax and caspase-3,cell apoptosis rate were all increased(P<0.05);Compared with the si-SNHG14+anti-NC group,the expression levels of miR-449a,Bax and caspase-3,and cell apoptosis rate in the si-SNHG14+anti miR-449a group were all decreased,whereas the expression levels of CCNE2,Cyclin D1 and Ki67,positive rate of Edu,scratch healing rate,

关 键 词：前列腺肿瘤 RNA 长链非编码 微RNAS 细胞周期蛋白E 裸小鼠 

分 类 号：R737.25[医药卫生—肿瘤]