Department of Stomatology Center, Shenzhen Hospital, Southern Medical University, Shenzhen, Guangdong 518100, P.R. China.
Mol Med Rep. 2021 Mar;23(3). doi: 10.3892/mmr.2020.11811. Epub 2021 Jan 5.
MicroRNAs (miRs) exhibit oncogenic or tumor suppressive functions that contribute to the initiation and development of various types of human cancer. miR‑149‑3p has been reported to serve multiple roles in the regulation of proliferation, apoptosis and metastasis. However, the effects and detailed mechanism of miR‑149‑3p in oral squamous cell carcinoma (OSCC) remain unclear. In the present study, miR‑149‑3p mimic, mimic control, miR‑149‑3p inhibitor and inhibitor control were transiently transfected into Cal27 and SCC‑9 cells. The viability, proliferation and apoptosis of OSCC cells were determined using Cell Counting Kit‑8, colony formation and Annexin V assays, respectively. The mRNA expression levels of miR‑149‑3p and AKT2 were determined by reverse transcription‑quantitative PCR. The protein expression levels of AKT2, cleaved caspase‑3 and cleaved PARP were examined by western blot analysis. The binding of miR‑149‑3p to the AKT2 3'‑untranslated region was evaluated by a dual luciferase reporter assay. In the present study, overexpression of miR‑149‑3p reduced the viability and proliferation of OSCC cells. By contrast, increased cell viability and proliferation was observed in miR‑149‑3p‑deficient OSCC cells. Dual luciferase reporter assay indicated that miR‑149‑3p significantly decreased the luciferase activity of the wild‑type AKT2 3'‑untranslated region. Moreover, overexpression of miR‑149‑3p downregulated the mRNA and protein expression levels of AKT2, suggesting that miR‑149‑3p was a negative modulator of AKT2. Restoration of AKT2 efficiently reversed the miR‑149‑3p‑mediated reduction in the proliferative capacity of OSCC cells. In addition, miR‑149‑3p enhanced the sensitivity of OSCC cells to the chemotherapeutic drug 5‑fluorouracil. Taken together, the current findings revealed an inhibitory effect of miR‑149‑3p on the proliferation of OSCC cells through the post‑transcriptional suppression of AKT2, and indicated a potential chemosensitizing function of miR‑149‑3p for the treatment of patients with OSCC.
微小 RNA（miRs）具有致癌或肿瘤抑制功能，有助于各种类型人类癌症的发生和发展。已经报道 miR-149-3p 在调节增殖、凋亡和转移方面具有多种作用。然而，miR-149-3p 在口腔鳞状细胞癌（OSCC）中的作用和详细机制尚不清楚。在本研究中，瞬时转染 miR-149-3p 模拟物、模拟物对照、miR-149-3p 抑制剂和抑制剂对照到 Cal27 和 SCC-9 细胞中。通过 Cell Counting Kit-8、集落形成和 Annexin V 测定分别确定 OSCC 细胞的活力、增殖和凋亡。通过逆转录-定量 PCR 测定 miR-149-3p 和 AKT2 的 mRNA 表达水平。通过 Western blot 分析检测 AKT2、cleaved caspase-3 和 cleaved PARP 的蛋白表达水平。通过双荧光素酶报告基因检测评估 miR-149-3p 与 AKT2 3'UTR 的结合。在本研究中，miR-149-3p 的过表达降低了 OSCC 细胞的活力和增殖。相反，在 miR-149-3p 缺陷的 OSCC 细胞中观察到细胞活力和增殖增加。双荧光素酶报告基因检测表明，miR-149-3p 显著降低了野生型 AKT2 3'UTR 的荧光素酶活性。此外，miR-149-3p 的过表达下调了 AKT2 的 mRNA 和蛋白表达水平，表明 miR-149-3p 是 AKT2 的负调节剂。AKT2 的恢复有效地逆转了 miR-149-3p 介导的 OSCC 细胞增殖能力的降低。此外，miR-149-3p 增强了 OSCC 细胞对化疗药物 5-氟尿嘧啶的敏感性。总之，目前的研究结果揭示了 miR-149-3p 通过 AKT2 的转录后抑制对 OSCC 细胞增殖的抑制作用，并表明 miR-149-3p 具有治疗 OSCC 患者的潜在化疗增敏作用。
