摘要
探讨miR-506靶向HOXA13对鼻咽癌细胞增殖、迁移和侵袭的影响。通过实时荧光定量PCR(RTq PCR)检测鼻咽上皮细胞NP69以及鼻咽癌细胞系CNE-1,CNE-2,HNE-1,C666-1中miR-506以及HOXA13 mRNA表达水平,选择CNE-1细胞用于后续实验,通过脂质体2000将miR-506 mimic、mimic-NC、siRNA-HOXA13、siRNANC转染CNE-1细胞,分为miR-506 mimic组、mimic-NC组、si-HOXA13组、si-NC组,对照组细胞不处理。通过RTq PCR检测各组细胞转染效率,CCK8检测各组细胞增殖情况,Transwell和划痕实验检测各组细胞侵袭和迁移情况,Western-blot实验检测各组细胞HOXA13、Ki-67、MMP-2蛋白表达水平,双荧光素酶实验验证靶向关系。与鼻咽癌上皮细胞NP69比,鼻咽癌细胞系中miR-506表达量显著下降(F=74.007,P<0.05),HOXA13 mRNA表达量显著升高(F=31.140,P<0.05)。与mimic-NC组和对照组比,miR-506 mimic组细胞增殖、侵袭和迁移能力显著下降,相关蛋白Ki-67、MMP-2蛋白表达量显著下降(P<0.05)。与si-NC组和对照组比,si-HOXA13组细胞增殖、侵袭和迁移能力显著下降,相关蛋白Ki-67、MMP-2蛋白表达量显著下降(P<0.05)。miR-506与HOXA13-3’UTR存在结合位点,双荧光素酶实验结果显示:HOXA13是miR-506的靶基因。miR-506通过靶向HOXA13抑制鼻咽癌细胞CNE-1增殖、侵袭和迁移,表明miR-506与鼻咽癌的发生发展密切相关。
To investigate the effects of miR-506 on the proliferation,migration and invasion of nasopharyngeal carcinoma cells by targeting HOXA13,quantitative real-time polymerase chain reaction(RT-qPCR)was used to detect expressions of miR-506 and HOXA13 mRNA in NPC cell line(CNE-1,CNE-2,HNE-1 and C666-1)and immortalized nasopharyngeal epithelial cells line(NP69).In the lowest-expressing nasopharyngeal carcinoma cell lines,the CNE-1 cells were transfected with miR-506 mimics,mimic-NC,siRNA-HOXA13 and siRNA-NC by liposome 2000,and the cells were divided into miR-506 mimic group,mimic-NC group,si-HOXA13 group and si-NC group.The control group cells were not treated.Real-time quantitative PCR was used to detect the efficiency of cell transfection.CCK-8 was used to detect cell proliferation.Transwell assay and Scratch test were used to detect cell invasion and migration.The protein expression levels of HOXA13,Ki-67 and MMP-2 were detected by Western-blot assay.Double luciferase assay was used to verify the targeted relationship.Compared with NP69 cell,the expression of miR-506 in NPC cell line was significantly decreased(F=74.007,P<0.05),and the expression of HOXA13 mRNA was significantly increased(F=31.140,P<0.05).Compared with the mimic-NC group and the control group,the bility of proliferation,invasion and migration of miR-506 mimic group and the protein expression levels of HOXA13,Ki-67 and MMP-2 were lower(P<0.05).Compared with the si-NC group and the control group,the bility of proliferation,invasion and migration of si-HOXA13 groups and the protein expression levels of HOXA13,Ki-67 and MMP-2 were lower(P<0.05).miR-506 had binding sites with HOXA13-3’UTR,and the results of double luciferase assay showed that HOXA13 was a target gene of miR-506.miR-506 inhibited the proliferation,invasion and migration of nasopharyngeal carcinoma cell CNE-1 by targeting HOXA13,indicating that miR-506 was closely related to the occurrence and development of nasopharyngeal carcinoma.
作者
黄妙铭
胡金成
毛志强
何锦添
HUANG Miao-ming;HU Jin-cheng;MAO Zhi-qiang;HE Jin-tian(Department of Otolaryngology,Dongguan People's Hospital,Dongguan 523059,China)
出处
《药物生物技术》
CAS
2021年第4期351-356,共6页
Pharmaceutical Biotechnology
基金
2020年东莞市社会科技发展(一般)项目(No.202050715001775)。
