Department of Otolaryngology, Qingpu Branch of Zhongshan Hospital, Fudan University, Shanghai, China.
Department of Otolaryngology, Zhongshan Hospital, Fudan University, Shanghai, China.
Neoplasma. 2018 Sep 19;65(5):730-735. doi: 10.4149/neo_2018_171119N752. Epub 2018 Sep 4.
miRNAs are among the most important factors that regulate gene expression. According to bioinformatic analysis, miR-365a-3p was predicted to interact with the TET1 mRNA. We predicted that it might affect tumor biological processes through TET1. TET1 interference and miR-365a-3p inhibitor constructs were generated. qRT-PCR was used to verify the expression level of miR-365a-3p and TET1 in Hep-2 and BESB-2B cells. qRT-PCR and Western blot were used to confirm the TET1 expression level in Hep-2 and miR-365a-3p inhibitor cells. Cell proliferation, cell cycle progression and cell invasion were further studied to identify the relationship between TET1 and miR-365a-3p. Luciferase reporter gene assays were used to find the binding site of miR-365a-3p in the 3'-UTR (3'-untranslated region) of the TET1 mRNA. TET1 was weakly expressed in Hep-2 cells and highly expressed in BESB-2B cells, while miR-143-3p and miR-365a-3p were highly expressed in Hep-2 cells and lowly expressed in BESB-2B cells. Inhibiting miR-365a-3p could up-regulate the expression of TET1. The negative effects of miR-365a-3p on cell proliferation, cell cycle progression and cell invasion could be abolished by TET1 interference. The binding site of miR-365a-3p was in the 3'-UTR of the TET1 mRNA. TET1 is one of the targets of miR-365a-3p. miR-365a-3p regulates the biological behavior of laryngeal cancer by down-regulating TET1.
miRNAs 是调节基因表达的最重要因素之一。根据生物信息学分析，miR-365a-3p 被预测与 TET1 mRNA 相互作用。我们预测它可能通过 TET1 影响肿瘤的生物过程。生成了 TET1 干扰和 miR-365a-3p 抑制剂构建体。使用 qRT-PCR 验证 Hep-2 和 BESB-2B 细胞中 miR-365a-3p 和 TET1 的表达水平。使用 qRT-PCR 和 Western blot 验证 Hep-2 和 miR-365a-3p 抑制剂细胞中的 TET1 表达水平。进一步研究细胞增殖、细胞周期进程和细胞侵袭，以确定 TET1 与 miR-365a-3p 之间的关系。使用荧光素酶报告基因检测法寻找 miR-365a-3p 在 TET1 mRNA 3'-UTR（3'-非翻译区）中的结合位点。TET1 在 Hep-2 细胞中表达较弱，在 BESB-2B 细胞中表达较高，而 miR-143-3p 和 miR-365a-3p 在 Hep-2 细胞中表达较高，在 BESB-2B 细胞中表达较低。抑制 miR-365a-3p 可以上调 TET1 的表达。TET1 干扰可以消除 miR-365a-3p 对细胞增殖、细胞周期进程和细胞侵袭的负性影响。miR-365a-3p 的结合位点位于 TET1 mRNA 的 3'-UTR。TET1 是 miR-365a-3p 的靶标之一。miR-365a-3p 通过下调 TET1 调节喉癌细胞的生物学行为。
