Department of Oncology, Liaocheng Infectious Disease Hospital, Liaocheng, China.
Eur Rev Med Pharmacol Sci. 2020 Sep;24(18):9429-9437. doi: 10.26355/eurrev_202009_23028.
The aim of this study was to investigate the effect of long non-coding RNA (lncRNA) WT-AS on the invasiveness and migration of non-small cell lung cancer (NSCLC) cells, and to explore the underlying molecular mechanism of lncRNA WT-AS in the pathogenesis of NSCLC.
LncRNA WT-AS expression in 50 pairs of NSCLC tissues and adjacent ones was studied by quantitative real-time polymerase chain reaction (qRT-PCR) analysis, and the correlations of WT-AS with clinicopathological indicators and prognosis of NSCLC patients were analyzed. Meanwhile, NSCLC expression levels in NSCLC cell lines were also evaluated by qPCR assay. In addition, WT-AS overexpression and knockdown models were constructed using lentivirus in NSCLC cell lines A549 and H1299, respectively. Thereafter, transwell and cell wound healing assays were carried out to assess the implication of WT-AS in biological functions of NSCLC cells. Furthermore, the interaction between WT-AS and KLK13 was determined via Luciferase assay.
The results showed that WT-AS expression in NSCLC was remarkably lower than that in normal tissues adjacent to the cancer. Univariate analysis suggested that compared with patients with high expression of WT-AS, patients in low expression group showed higher incidence of metastasis and lower survival rates. Overexpression of WT-AS suppressed cell invasion and metastasis capacity, while the opposite result was observed in WT-AS knockdown group. KLK13 expression showed an increase in NSCLC cell lines and tissues, which was negatively correlated with WT-AS level. Meanwhile, Luciferase assay confirmed the binding between WT-AS and KLK13. Western blotting revealed that KLK13 expression was remarkably elevated in EC tissues and was positively correlated with TRIM62. In addition, it was also found that WT-AS and KLK13 had a mutual regulatory effect, which together affect the malignant progress of NSCLC.
This study shows for the first time that LncRNA WT-AS interacts with KLK13 to serve as a negative regulator of NSCLC progression.
本研究旨在探讨长链非编码 RNA（lncRNA）WT-AS 对非小细胞肺癌（NSCLC）细胞侵袭和迁移的影响，并探讨 lncRNA WT-AS 在 NSCLC 发病机制中的潜在分子机制。
采用实时定量聚合酶链反应（qRT-PCR）分析 50 对 NSCLC 组织及其相邻组织中 lncRNA WT-AS 的表达，并分析 WT-AS 与 NSCLC 患者临床病理指标和预后的相关性。同时，通过 qPCR 检测 NSCLC 细胞系中 NSCLC 的表达水平。此外，分别通过慢病毒在 NSCLC 细胞系 A549 和 H1299 中构建 WT-AS 过表达和敲低模型。然后，通过 Transwell 和细胞划痕愈合实验评估 WT-AS 对 NSCLC 细胞生物学功能的影响。此外，通过荧光素酶测定法确定 WT-AS 与 KLK13 之间的相互作用。
结果表明，WT-AS 在 NSCLC 中的表达明显低于癌旁正常组织。单因素分析表明，与 WT-AS 高表达患者相比，低表达组患者转移发生率更高，生存率更低。过表达 WT-AS 抑制细胞侵袭和转移能力，而在 WT-AS 敲低组则观察到相反的结果。KLK13 在 NSCLC 细胞系和组织中的表达增加，与 WT-AS 水平呈负相关。同时，荧光素酶测定法证实了 WT-AS 与 KLK13 的结合。Western blot 显示 KLK13 在 EC 组织中表达显著升高，并与 TRIM62 呈正相关。此外，还发现 WT-AS 和 KLK13 存在相互调节作用，共同影响 NSCLC 的恶性进展。
本研究首次表明，lncRNA WT-AS 与 KLK13 相互作用，作为 NSCLC 进展的负调节剂。
