Department of Stomatology, Beijing Luhe Hospital, Capital Medical University, Beijing, China.
Dev Growth Differ. 2019 Dec;61(9):466-474. doi: 10.1111/dgd.12638. Epub 2019 Nov 21.
Long non-coding RNAs (lncRNAs) play essential roles in the regulation of gene transcription in carcinogenesis and metastasis via interacting with microRNA. In this study, we explored the expressions and functions of lncRNA KCNQ1OT1 and miR-185-5p in oral squamous cell carcinoma (OSCC) cells. KCNQ1OT1 expression in OSCC tissues and cells was examined by qRT-PCR. Small interfering RNAs against KCNQ1OT1 (si- KCNQ1OT1) were used to knockdown KCNQ1OT1 in OSCC cells. Cell function was assessed by wound healing assay, transwell assay, and apoptosis detection. The binding region between KCNQ1OT1 and miR-185-5p was confirmed by luciferase assays. MiR-185-5p expression was measured by qRT-PCR. Rab14 was confirmed as a downstream target gene of miR-185-5p by measuring luciferase activities. The protein level of Rab14 in OSCC cells transfected with miR-185-5p or si-KCNQ1OT1 was determined by Western blot. The OSCC cell function and Rab14 expression after co-transfection of si-KCNQ1OT1 and miR-185-5p inhibitor were also examined. KCNQ1OT1 was upregulated in OSCC tissues and cells. KCNQ1OT1 silencing suppressed OSCC cell malignancy and downregulated miR-185-5p level, which showed upregulated expression in OSCC samples. Rab14 as a target gene of miR-185-5p was highly expressed in OSCC. KCNQ1OT1 knockdown impaired the invasion capability of OSCC cells, promoted apoptosis, and suppressed Rab14 expression. The inhibition of miR-185-5p in KCNQ1OT1 silencing cells reversed the suppression of Rab14 and restored the cancerous growth of OSCC cells. These results indicated that KCNQ1OT1 promoted OSCC tumorigenesis via the modulation of miR-185-5p/Rab14 axis, which may serve as a therapeutic target for the treatment of OSCC.
长链非编码 RNA（lncRNA）通过与 microRNA 相互作用，在致癌和转移过程中发挥着重要作用，调节基因转录。在这项研究中，我们探讨了 lncRNA KCNQ1OT1 和 miR-185-5p 在口腔鳞状细胞癌（OSCC）细胞中的表达和功能。通过 qRT-PCR 检测 OSCC 组织和细胞中的 KCNQ1OT1 表达。用针对 KCNQ1OT1 的小干扰 RNA（si-KCNQ1OT1）转染 OSCC 细胞，以敲低 KCNQ1OT1。通过划痕愈合试验、Transwell 试验和凋亡检测评估细胞功能。通过荧光素酶报告基因实验证实 KCNQ1OT1 和 miR-185-5p 之间的结合区域。通过 qRT-PCR 测量 miR-185-5p 的表达。通过测量荧光素酶活性证实 Rab14 是 miR-185-5p 的下游靶基因。Western blot 测定转染 miR-185-5p 或 si-KCNQ1OT1 的 OSCC 细胞中 Rab14 的蛋白水平。还检测了共转染 si-KCNQ1OT1 和 miR-185-5p 抑制剂后 OSCC 细胞功能和 Rab14 表达的变化。KCNQ1OT1 在 OSCC 组织和细胞中上调。KCNQ1OT1 沉默抑制 OSCC 细胞恶性程度，并下调 miR-185-5p 水平，OSCC 样本中显示上调表达。Rab14 作为 miR-185-5p 的靶基因在 OSCC 中高表达。KCNQ1OT1 敲低抑制 OSCC 细胞侵袭能力，促进凋亡，并抑制 Rab14 表达。在 KCNQ1OT1 沉默细胞中抑制 miR-185-5p 可逆转 Rab14 的抑制作用，并恢复 OSCC 细胞的癌变生长。这些结果表明，KCNQ1OT1 通过调节 miR-185-5p/Rab14 轴促进 OSCC 肿瘤发生，可能成为治疗 OSCC 的治疗靶点。
