目的：分析circRHOT1、miR-144-3p在喉癌组织中的表达水平,探讨circRHOT1靶向miR-144-3p对喉癌细胞生物学行为的影响。方法：RT-qPCR检测circRHOT1、miR-144-3p在32例喉癌组织标本和15例声带息肉组织标本中表达情况。Pearson相关分析确定喉癌组织中circRHOT1、miR-144-3p表达关系。将si-NC、si-circRHOT1、miR-NC、miR-144-3p mimics、si-circRHOT1+ anti-miR-NC、si-circRHOT1+anti-miR-144-3p分别转染TU686细胞,采用CCK-8法、划痕愈合实验、流式细胞术分析TU686细胞的体外增殖、迁移能力和凋亡情况。双荧光素酶报告实验用于分析circRHOT1、miR-144-3p的靶向关系。结果：喉癌组织中circRHOT1表达水平较声带息肉组织显著升高,miR-144-3p表达水平较声带息肉组织显著降低。喉癌组织中circRHOT1、miR-144-3p表达呈负相关关系。下调circRHOT1表达后TU686细胞抑制率、凋亡率、miR-144-3p表达水平显著升高,划痕愈合率显著降低。过表达miR-144-3p后TU686细胞抑制率、凋亡率显著升高,划痕愈合率显著降低。miR-144-3p是circRHOT1的直接靶点。抑制miR-144-3p表达可减弱下调circRHOT1对TU686细胞增殖、凋亡和迁移的影响。结论：下调circRHOT1通过靶向上调miR-144-3p表达可诱导喉癌细胞凋亡,抑制其增殖和迁移。
Abstract
Objective To analyze the expression levels of circRHOT1 and miR-144-3p in laryngeal cancer tissues, and further to explore the effect of circRHOT1 targeting miR-144-3p on the biological behavior of laryngeal cancer cells. Methods circRHOT1 and miR-144-3p expression in 32 specimens of laryngeal cancer and 15 specimens of vocal cord polyp tissues was estimated by RT-qPCR. Pearson correlation analysis was used to determined the expression relationship of circRHOT1 and miR-144-3p in laryngeal cancer tissues. si-NC, si-circRHOT1, miR-NC, miR-144-3p mimics, si-circRHOT1+anti-miR-NC, si-circRHOT1+anti-miR-144-3p were respectively transfected into TU686 cells. CCK-8 method, scratch healing assays and flow cytometry were applied to analyze the proliferation, migration ability and apoptosis of TU686 cell in vitro. Dual luciferase reporter assay was used to analyze the targeting relationship between circRHOT1 and miR-144-3p. Results The expression level of circRHOT1 in laryngeal cancer tissue was significantly higher than that in vocal cord polyp tissues, while the expression level of miR-144-3p was significantly lower than that in vocal cord polyp tissues. The expression of circRHOT1 and miR-144-3p in laryngeal cancer tissues was negatively correlated. After down-regulating the expression of circRHOT1, the inhibition rate, apoptosis rate and miR-144-3p expression level of TU686 cells were significantly increased, and the scratch healing rate was significantly reduced. After overexpression of miR-144-3p, the inhibition rate and apoptosis rate of TU686 cells were significantly increased, and the scratch healing rate was significantly reduced. miR-144-3p was considered as a direct target of circRHOT1. Inhibition of miR-144-3p could attenuate the effect of circRHOT1 down-regulation on the proliferation, apoptosis and migration of TU686 cells. Conclusion Down-regulation of circRHOT1 could induce laryngeal cancer cell apoptosis and inhibit their proliferation and migration by targeting and up-regulating miR-144-3p expression.
关键词
喉癌 /
circRHOT1 /
miR-144-3p /
细胞增殖 /
迁移 /
凋亡
Key words
laryngeal carcinoma /
circRHOT1 /
miR-144-3p /
cell proliferation /
migration /
apoptosis
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基金
河北省保定市科技计划项目（2141ZF163）
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