Chen Xiaohua, Li Xiaohua, Qin Zhaobing
a Otolaryngology Department , First Affiliated Hospital of Zhengzhou University , Zhengzhou , Henan , PR China.
Acta Otolaryngol. 2016 Dec;136(12):1261-1266. doi: 10.1080/00016489.2016.1202447. Epub 2016 Jul 4.
The present study revealed that miR-21 promotes the proliferation and invasion of cholesteatoma keratinocytes. These results provide a partial explanation for the more aggressive clinical behavior observed in cholesteatoma.
This study aims to investigate the post-transcriptional regulatory effects that control proliferation, apoptosis, and invasion in cholesteatoma keratinocytes. In particular, the potential role of miR-21 was focused on in this study.
Thirty cholesteatoma tissues were processed for RNA and cell culture. Cholesteatoma keratinocytes were transfected with miR-21 mimics, miR-21 inhibitors, or negative control miRNAs; and growth curves were drawn. RT-PCR was used to assess the expression levels of miR-21. EdU incorporation assay and TUNEL staining were used to assess the proliferation and apoptosis of cholesteatoma keratinocytes, respectively. The invasive abilities of cholesteatoma keratinocytes were examined using 6-well Transwell plates.
MiRNA-21 was upregulated when cholesteatoma keratinocytes were transfected with miR-21 mimics. Furthermore, the number of proliferative EdU + cells increased in cholesteatoma keratinocytes transfected with miR-21 mimics; and the number of TUNEL-positive cells also increased in cells transfected with miR-21 mimics. In addition, the number of migrated cells transfected with miR-21 mimics was higher, compared with migrated cells transfected miR-21 inhibitors or control miRNA.
本研究表明,miR-21促进胆脂瘤角质形成细胞的增殖和侵袭。这些结果为胆脂瘤中观察到的更具侵袭性的临床行为提供了部分解释。
本研究旨在探讨控制胆脂瘤角质形成细胞增殖、凋亡和侵袭的转录后调控作用。本研究特别关注了miR-21的潜在作用。
对30个胆脂瘤组织进行RNA提取和细胞培养。用miR-21模拟物、miR-21抑制剂或阴性对照miRNA转染胆脂瘤角质形成细胞,并绘制生长曲线。采用RT-PCR评估miR-21的表达水平。分别采用EdU掺入法和TUNEL染色法评估胆脂瘤角质形成细胞的增殖和凋亡。使用6孔Transwell板检测胆脂瘤角质形成细胞的侵袭能力。
用miR-21模拟物转染胆脂瘤角质形成细胞时,miRNA-21上调。此外,用miR-21模拟物转染的胆脂瘤角质形成细胞中增殖的EdU+细胞数量增加;用miR-21模拟物转染的细胞中TUNEL阳性细胞数量也增加。此外,与用miR-21抑制剂或对照miRNA转染的迁移细胞相比,用miR-21模拟物转染的迁移细胞数量更多。