ABSTRACT

The function of a new long non-coding RNA GAS6-AS2 in non-small cell lung cancer (NSCLC) is not fully understood. In this study, GAS6-AS2 was identified, and its roles as well as mechanisms in regulating proliferation of NSCLCs cells were investigated. qRT-PCR was used to analyze GAS6-AS2, miR-144-3p, and MAPK6 expression. Protein expression was detected by Western blotting. Cell Counting Kit-8 (CCK8) assay was used to examine the cell proliferation ability. The interaction between GAS6-AS2 and miR-144-3p was confirmed by dual-luciferase reporter assay and RNA pull down assay. A xenograft model was constructed to monitor the mice NSCLC tumor growth in vivo. GAS6-AS2 was up-regulated, while miR-144-3p was suppressed in NSCLC cells compared with normal lung cells. GAS6-AS2 suppression could inhibit the progression of NSCLC cells, and miR-144-3p could attenuate the effect. GAS6-AS2 could function as a competitive endogenous RNA (ceRNA) via direct sponging miR-144-3p-3p, which further regulating the expression of MAPK6. The knockdown of GAS6-AS2 could greatly suppress the tumor growth of NSCLC in vivo. GAS6-AS2 up-regulated MAPK6 by sponging miR-144-3p in NSCLC tissues and cells. Thus, GAS6-AS2 is an effective therapeutic target in NSCLC.

摘要

一种新的长链非编码 RNA GAS6-AS2 在非小细胞肺癌 (NSCLC) 中的功能尚不完全清楚。本研究鉴定了 GAS6-AS2，并研究了其在调节 NSCLC 细胞增殖中的作用和机制。qRT-PCR 用于分析 GAS6-AS2、miR-144-3p 和 MAPK6 的表达。通过蛋白质印迹检测蛋白质表达。Cell Counting Kit-8 (CCK8) 测定用于检查细胞增殖能力。GAS6-AS2 和 miR-144-3p 之间的相互作用通过双荧光素酶报告基因分析和 RNA 下拉分析得到证实。构建异种移植模型以监测小鼠体内 NSCLC 肿瘤的生长. 与正常肺细胞相比，非小细胞肺癌细胞中的 GAS6-AS2 被上调，而 miR-144-3p 被抑制。GAS6-AS2抑制可以抑制NSCLC细胞的进展，而miR-144-3p可以减弱这种作用。GAS6-AS2 可以通过直接海绵化 miR-144-3p-3p 发挥竞争性内源性 RNA (ceRNA) 的作用，从而进一步调节 MAPK6 的表达。GAS6-AS2的敲低可以极大地抑制体内NSCLC的肿瘤生长。GAS6-AS2 通过在 NSCLC 组织和细胞中形成 miR-144-3p 上调 MAPK6。因此，GAS6-AS2 是 NSCLC 的有效治疗靶点。