UNASSIGNED: The transcriptional levels of THAP9-AS1 and miR-185-5p were estimated via quantitative real time polymerase chain reaction (qRT-PCR) assay. The protein level of SOX13 was detected with western blotting assay. Additionally, methyl thiazolyl tetrazolium (MTT) assay as well as colony formation assay were utilized to measure cell growth. The apoptotic cells were observed by employing Terminal-deoxynucleoitidyl Transferase Mediated Nick End Labeling (TUNEL) staining analysis, and transwell assay was introduced to test cell migration in addition to invasion. Moreover, the relationship between miR-185-5p and THAP9-AS1 or SOX13 was estimated through dual-luciferase reporter gene assay.
UNASSIGNED: THAP9-AS1 was overexpressed in head and neck squamous cell carcinoma (HNSCC) tissues and NPC cells. Besides, silencing of THAP9-AS1 depressed the life processes of NPC cells including cell growth, migration as well as invasion but facilitated cell apoptosis. Further investigation proved that miR-185-5p was the direct target of THAP9-AS1. Besides, the knockdown of THAP9-AS1 notably reduced the transcriptional level of miR-185-5p. Furthermore, THAP9-AS1 served as a sponge of miR-185-5p to modulate the expression of SOX13, which regulated the development of NPC cells.
UNASSIGNED: This work verified that THAP9-AS1 promoted NPC cell progression at least partly by mediating the miR-185-5p/SOX13 axis.
■通过定量实时聚合酶链反应(qRT-PCR)测定来估计THAP9-AS1和miR-185-5p的转录水平。用蛋白质印迹法检测SOX13的蛋白质水平。此外,甲基噻唑基四唑(MTT)测定法以及集落形成测定法用于测量细胞生长。通过使用末端脱氧核苷酸转移酶介导的尼克末端标记(TUNEL)染色分析观察凋亡细胞。并引入transwell测定法来测试细胞迁移以及侵袭。此外,miR-185-5p与THAP9-AS1或SOX13之间的关系是通过双荧光素酶报告基因测定来评估的.
■THAP9-AS1在头颈部鳞状细胞癌(HNSCC)组织和NPC细胞中过表达。此外,沉默THAP9-AS1抑制了NPC细胞的生命过程,包括细胞生长,迁移以及侵袭,但促进细胞凋亡。进一步研讨证明miR-185-5p是THAP9-AS1的直接靶点。此外,THAP9-AS1的敲除显著降低了miR-185-5p的转录水平。此外,THAP9-AS1充当miR-185-5p的海绵来调节SOX13的表达,从而调节NPC细胞的发育。
■这项工作验证了THAP9-AS1至少部分地通过介导miR-185-5p/SOX13轴来促进NPC细胞进展。