Department of Dental Surgery, Yantaishan Hospital, Yantai, China.
Eur Rev Med Pharmacol Sci. 2020 May;24(9):4803-4809. doi: 10.26355/eurrev_202005_21169.
The aim of this study was to investigate the role of micro ribonucleic acid (miR)-92a in the pathogenesis of oral squamous cell carcinoma (OSCC).
The relative expression level of miR-92a in OSCC cell lines and normal oral epithelial keratinocyte cell lines was detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Subsequently, the effects of miR-92a silencing on proliferation and apoptosis of OSCC cells were determined via cell counting kit-8 (CCK-8) assay, flow cytometry, and colony formation assay, respectively. Whether miR-92a could bind to the 3'-untranslated region (3'-UTR) of target mRNA was detected using Dual-Luciferase reporter gene assay. Furthermore, changes in Wnt/β-catenin pathway-associated proteins were explored via Western blotting.
The expression of miR-92a in OSCC cell lines was significantly higher than that of normal oral epithelial keratinocyte cell lines (p<0.05). Silencing of miR-92a significantly inhibited proliferation and promoted apoptosis of OSCC cells (p<0.05). Luciferase reporter gene assay confirmed that miR-92a could bind to the 3'-UTR of Kruppel-like factor 4 (KLF4) gene. After miR-92a silencing, the expressions of Wnt/β-catenin pathway-associated proteins were remarkably down-regulated, including β-catenin, c-Myc, and Wnt3a (p<0.05).
Silencing of miR-92a inactivates the Wnt/β-catenin signaling pathway by targeting KLF4, thereby inhibiting proliferation and promoting apoptosis of OSCC cells. Our findings suggest that miR-92a may be a potential therapeutic target for OSCC patients.
本研究旨在探讨微小 RNA-92a（miR-92a）在口腔鳞状细胞癌（OSCC）发病机制中的作用。
采用实时定量逆转录聚合酶链反应（qRT-PCR）检测 miR-92a 在 OSCC 细胞系和正常口腔上皮角质形成细胞系中的相对表达水平。随后，通过细胞计数试剂盒-8（CCK-8）检测、流式细胞术和集落形成实验分别检测 miR-92a 沉默对 OSCC 细胞增殖和凋亡的影响。采用双荧光素酶报告基因检测法检测 miR-92a 是否能与靶 mRNA 的 3′非翻译区（3′UTR）结合。此外，通过 Western blot 探讨 Wnt/β-catenin 通路相关蛋白的变化。
OSCC 细胞系中 miR-92a 的表达明显高于正常口腔上皮角质形成细胞系（p<0.05）。沉默 miR-92a 可显著抑制 OSCC 细胞的增殖并促进其凋亡（p<0.05）。荧光素酶报告基因检测证实 miR-92a 可与 Kruppel 样因子 4（KLF4）基因的 3′UTR 结合。沉默 miR-92a 后，Wnt/β-catenin 通路相关蛋白表达显著下调，包括β-catenin、c-Myc 和 Wnt3a（p<0.05）。
沉默 miR-92a 通过靶向 KLF4 使 Wnt/β-catenin 信号通路失活，从而抑制 OSCC 细胞的增殖并促进其凋亡。研究结果提示，miR-92a 可能成为 OSCC 患者的潜在治疗靶点。
