CDKN2B-AS1通过调控miR-98-5p/E2F2轴促进鼻咽癌细胞的增殖、克隆形成及侵袭。
CDKN2B-AS1 promotes the proliferation, clone formation, and invasion of nasopharyngeal carcinoma cells by regulating miR-98-5p/E2F2 axis.
作者信息
Li Zhengwen, Cai Xiaojing, Zou Wentao, Zhang Jiaxiong
机构信息
Department of Otorhinolaryngology, Shanghai Tenth Peoples' Hospital, Tongji University Shanghai 200072, China.
出版信息
Am J Transl Res. 2021 Dec 15;13(12):13406-13422. eCollection 2021.
OBJECTIVE
To explore the effect of CDKN2B antisense RNA 1 (CDKN2B-AS1) on the proliferation, clone formation, and invasion of nasopharyngeal carcinoma (NPC) cells by regulating miR-98-5p/E2F transcription factor 2 (E2F2) axis.
METHODS
The expressions of CDKN2B-AS1, miR-98-5p, and E2F2 in NPC tissues and cell lines (SUNE-1, 5-8F, 6-10B, and HK-1) as well as in peritumoral normal tissues and cell line NP69 were determined by qRT-PCR. Subcellular localization of CDKN2B-AS1 was detected using the fluorescence hybridization assay. The targeting relationships between CDKN2B-AS1 and miR-98-5p as well as between miR-98-5p and E2F2 were analyzed by the dual-luciferase reporter assay and RNA binding protein immunoprecipitation assay. The proliferation, clone formation and invasion of 5-8F cells were measured using the CCK-8 assay, Clone formation assay, and transwell assay, respectively.
RESULTS
CDKN2B-AS1 was highly expressed in NPC tissues and cells, whereas the expression of miR-98-5p decreased in the NPC tissues and cells. Silencing of CDKN2B-AS1 inhibited the proliferation, clone formation, and invasion of NPC cells (all P<0.05). CDKN2B-AS1 acted asceRNA of miR-98-5p, and miR-98-5p inhibitor could partially reverse the inhibitory effect of silencing CDKN2B-AS1 on NPC cells (all P<0.05). CDKN2B-AS1 upregulated E2F2 by inhibiting miR-98-5p, and the upregulation of E2F2 partially reversed the inhibitory effect of miR-98-5p overexpression on the NPC cells (all P<0.05).
CONCLUSION
CDKN2B-AS1, as a lncRNA, can regulate E2F2 by sponging miR-98-5p to promote the proliferation, clone formation, and invasion of NPC cells.
目的
通过调节miR-98-5p/E2F转录因子2（E2F2）轴，探讨CDKN2B反义RNA 1（CDKN2B-AS1）对鼻咽癌（NPC）细胞增殖、克隆形成及侵袭的影响。
方法
采用qRT-PCR检测NPC组织和细胞系（SUNE-1、5-8F、6-10B和HK-1）以及癌旁正常组织和细胞系NP69中CDKN2B-AS1、miR-98-5p和E2F2的表达。利用荧光杂交试验检测CDKN2B-AS1的亚细胞定位。通过双荧光素酶报告基因试验和RNA结合蛋白免疫沉淀试验分析CDKN2B-AS1与miR-98-5p以及miR-98-5p与E2F2之间的靶向关系。分别采用CCK-8试验、克隆形成试验和Transwell试验检测5-8F细胞的增殖、克隆形成和侵袭能力。
结果
CDKN2B-AS1在NPC组织和细胞中高表达，而miR-98-5p在NPC组织和细胞中的表达降低。沉默CDKN2B-AS1可抑制NPC细胞的增殖、克隆形成及侵袭（均P<0.05）。CDKN2B-AS1作为miR-98-5p的ceRNA，miR-98-5p抑制剂可部分逆转沉默CDKN2B-AS1对NPC细胞的抑制作用（均P<0.05）。CDKN2B-AS1通过抑制miR-98-5p上调E2F2，E2F2的上调部分逆转了miR-98-5p过表达对NPC细胞的抑制作用（均P<0.05）。
结论
CDKN2B-AS1作为一种lncRNA，可通过吸附miR-98-5p来调节E2F2，从而促进NPC细胞的增殖、克隆形成及侵袭。
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