Cutaneous malignant melanoma (CMM) is responsible for ≥1/2 of skin cancer‑related mortalities. The aberrant expression of long non‑coding RNAs (lncRNAs) has been associated with the development of CMM. However, to the best of our knowledge, the role of the lncRNA TINCR ubiquitin domain containing (TINCR) in CMM has not been previously investigated, and thus, the current study aimed to evaluate this in vitro and in vivo. Reverse transcription‑quantitative PCR (RT‑qPCR) was used to analyze microRNA (miR)‑424‑5p expression, and RT‑qPCR and western blotting were used to measure TINCR, large tumor suppressor kinase 1 (LATS1), cellular communication network factor 2 (CTGF), cellular communication network factor 1 (CCN1) and AXL receptor tyrosine kinase (AXL) mRNA and protein expression levels, respectively. Cell Counting Kit‑8, flow cytometry and Transwell assays were used to detect the proliferation, apoptosis and invasion of CMM cell lines, respectively. The binding sites between TINCR and miR‑424‑5p were predicted using the miRDB database. A dual luciferase reporter assay and RT‑qPCR were used to identify the relationship between TINCR and miR‑424‑5p in CMM cell lines. The bioinformatics analysis revealed that TINCR was one of the most significantly downregulated lncRNAs in CMM, and advanced stage CMM tissues showed the greatest decrease in TINCR expression. Moreover, in the collected CMM tissues and tested cell lines of the current study, TINCR expression was found to be downregulated compared with the respective controls. Notably, TINCR overexpression inhibited the expression levels of CTGF, CCN1 and AXL, decreased the proliferation and invasion, and induced the apoptosis of CMM cell lines. In addition, a mutual binding association was identified between miR‑424‑5p and TINCR in CMM cells. LATS1, a target of miR‑424‑5p, was found to be positively regulated by TINCR. TINCR activated Hippo signaling and repressed the activity of Yes 1 associated transcriptional regulator by regulating LATS1 expression, while LATS1 knockdown reversed the effect of TINCR overexpression on CMM cells. Collectively, the findings of the present study suggested that TINCR may attenuate the progression of CMM by regulating the miR‑424‑5p/LATS1 signaling axis. These results indicated that TINCR may play a tumor suppressive role in CMM.

中文翻译：

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lncRNA TINCR 通过调节 miR-424-5p/LATS1 轴来减弱皮肤恶性黑色素瘤细胞的增殖和侵袭，并增强其凋亡。


皮肤恶性黑色素瘤 (CMM) 导致 ≥1/2 的皮肤癌相关死亡。长非编码 RNA (lncRNA) 的异常表达与 CMM 的发生有关。然而，据我们所知，之前尚未研究过含有 TINCR 泛素结构域的 lncRNA TINCR 在 CMM 中的作用，因此，本研究旨在体外和体内评估这一作用。采用逆转录定量 PCR (RT-qPCR) 分析 microRNA (miR)-424-5p 表达，采用 RT-qPCR 和蛋白质印迹法检测 TINCR、大肿瘤抑制激酶 1 (LATS1)、细胞通讯网络因子2 (CTGF)、细胞通讯网络因子 1 (CCN1) 和 AXL 受体酪氨酸激酶 (AXL) mRNA 和蛋白表达水平分别。采用Cell Counting Kit-8、流式细胞术和Transwell实验分别检测CMM细胞系的增殖、凋亡和侵袭。使用 miRDB 数据库预测 TINCR 和 miR-424-5p 之间的结合位点。使用双荧光素酶报告基因测定和 RT-qPCR 来确定 CMM 细胞系中 TINCR 和 miR-424-5p 之间的关系。生物信息学分析显示，TINCR是CMM中下调最显着的lncRNA之一，晚期CMM组织中TINCR表达下降幅度最大。此外，在本研究收集的CMM组织和测试的细胞系中，发现与各自的对照相比，TINCR表达下调。值得注意的是，TINCR过表达抑制CTGF、CCN1和AXL的表达水平，降低增殖和侵袭，并诱导CMM细胞系凋亡。 此外，在 CMM 细胞中，miR-424-5p 和 TINCR 之间存在相互结合关联。 LATS1 是 miR-424-5p 的靶标，被发现受到 TINCR 的正向调节。 TINCR 激活 Hippo 信号传导并通过调节 LATS1 表达抑制 Yes 1 相关转录调节因子的活性，而 LATS1 敲低则逆转了 TINCR 过表达对 CMM 细胞的影响。总的来说，本研究的结果表明 TINCR 可能通过调节 miR-424-5p/LATS1 信号轴来减弱 CMM 的进展。这些结果表明TINCR可能在CMM中发挥肿瘤抑制作用。