摘要
该文旨在探究长链非编码RNA CBR3-AS1(lncRNA CBR3-AS1)靶向微小RNA-145-5p(miR-145-5p)/肌动蛋白束蛋白1(FSCN1)轴对鼻咽癌(NPC)细胞增殖、凋亡和侵袭的影响。qRTPCR法检测鼻咽癌组织中lncRNACBR3-AS1和miR-145-5p的表达水平。将鼻咽癌细胞CNE-1分为si-NC组、si-CBR3-AS1组、si-CBR3-AS1+anti-miR-NC组、si-CBR3-AS1+anti-miR-145-5p组、miR-NC组、miR-145-5pmimics组、miR-145-5pmimics+pcDNA组、miR-145-5pmimics+FSCN1组。双荧光素酶实验检测lncRNA CBR3-AS1和miR-145-5p及FSCN1和miR-145-5p的靶向关系;MTT法检测细胞增殖情况;Annexin V-FITC/PI法检测细胞凋亡情况;Transwell实验检测细胞侵袭能力;Western blot检测细胞周期负调控因子(P21)、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)的蛋白表达变化。建立小鼠移植瘤模型,探究lncRNA CBR3-AS1对鼻咽癌移植瘤生长及miR-145-5p/FSCN1轴的影响。在鼻咽癌组织中,lncRNA CBR3-AS1表达上调,miR-145-5p表达下调(P<0.05)。干扰lncRNA CBR3-AS1或上调miR-145-5p表达可以抑制鼻咽癌细胞的增殖与侵袭,促进鼻咽癌细胞凋亡(P<0.05)。下调miR-145-5p表达或上调FSCN1表达可以逆转干扰lncRNA CBR3-AS1表达或上调miR-145-5p表达对鼻咽癌细胞的增殖与侵袭的抑制作用(P<0.05)。小鼠移植瘤实验显示,下调lncRNA CBR3-AS1表达可上调miR-145-5p表达,从而下调FSCN1表达,抑制鼻咽癌移植瘤的生长(P<0.05)。lncRNA CBR3-AS1在鼻咽癌组织中上调表达,下调lncRNA CBR3-AS1表达可通过调控miR-145-5p/FSCN1轴抑制鼻咽癌细胞的增殖与侵袭,促进细胞凋亡。
This study aims to investigate the impacts of lncRNA CBR3-AS1(long non-coding RNA CBR3-AS1)on the proliferation,apoptosis and invasion of NPC(nasopharyngeal carcinoma)cells by targeting the miR-145-5p(microRNA-145-5p)/FSCN1(fascin 1)axis.The qRT-PCR method was applied to analyze the expression levels of lncRNA CBR3-AS1 and miR-145-5p in nasopharyngeal carcinoma tissues.Nasopharyngeal cancer cell CNE-1 was divided into si-NC group,si-CBR3-AS1 group,si-CBR3-AS1+anti-miR-NC group,si-CBR3-AS1+anti-miR-145-5p group,miR-NC group,miR-145-5p mimics group,miR-145-5p mimics+pcDNA group,and miR-145-5p mimics+FSCN1 group.Double luciferase experiment was applied to detect the targeting relationship between lncRNA CBR3-AS1 and miR-145-5p,and between FSCN1 and miR-145-5p;MTT method was applied to detect cell proliferation;Annexin V-FITC/PI method was applied to detect cell apoptosis;Transwell experiment was applied to detect cell invasion ability;Western blot was applied to detect the protein expression changes of P21(cell cycle negative regulator),Bcl-2(B cell lymphoblastoma-2),Bax(Bcl-2 associated X protein),MMP-2(ma-trix metalloproteinase 2),and MMP-9(matrix metalloproteinase 9).Mouse transplantation tumor experiment was applied to explore the effect of LncRNA CBR3-AS1 on the growth of nasopharyngeal carcinoma transplantation tumors and miR-145-5p/FSCN1.In nasopharyngeal carcinoma tissue,the expression of lncRNA CBR3-AS1 was up-regulated,while the expression of miR-145-5p was down-regulated(P<0.05).Interference with lncRNA CBR3-AS1 or up-regulation of miR-145-5p expression was able to inhibit the proliferation and invasion of nasopharyngeal carcinoma cells,and promote the apoptosis of nasopharyngeal carcinoma cells(P<0.05).Down-regulation of miR-145-5p expression or up-regulation of FSCN1 expression was able to reverse the inhibitory effects of interfering with lncRNA CBR3-AS1 expression or up-regulation of miR-145-5p expression on the proliferation and invasion of nasopharyngeal carcinoma cells(P<0.05).The mouse transplantation tumor experiment showed that down-regu-lating the expression of lncRNA CBR3-AS1 was able to up-regulate the expression of miR-145-5p,thereby down-regulating the expression of FSCN1 and inhibiting the growth of nasopharyngeal carcinoma transplantation tumors(P<0.05).lncRNA CBR3-AS1 is up-regulated in nasopharyngeal carcinoma tissue,down-regulating the expression of lncRNA CBR3-AS1 can inhibit the proliferation and invasion of nasopharyngeal carcinoma cells,and promote cell apoptosis by regulating the miR-145-5p/FSCN1 axis.
作者
王华新
娄丹
杨磊
张海忠
刘丽莎
WANG Huaxin;LOU Dan;YANG Lei;ZHANG Haizhong;LIU Lisha(Department of Otolaryngology,the Sixth Hospital of Shijiazhuang,Shijiazhuang 050000,China;Department of Otolaryngology,Head and Neck Surgery,the First Hospital of Qinhuangdao,Qinhuangdao 066000,China;Department of Otolaryngology,the Second Hospital of Hebei Medical University,Shijiazhuang 050000,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2024年第3期515-525,共11页
Chinese Journal of Cell Biology
基金
2022年度河北省医学科学研究课题计划(批准号:20220198)资助的课题。
