Department of Thoracic Surgery, Cancer Hospital of University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Hangzhou, China.
Institute of Cancer and Basic Medicine (IBMC), Chinese Academy of Science, Hangzhou, China.
Bioengineered. 2021 Dec;12(1):3525-3538. doi: 10.1080/21655979.2021.1950282.
Lung cancer (LC) ranks first among all causes of cancer-related death, with non-small cell lung cancer (NSCLC) taking up 85% of lung cancer cases. Although lncRNA MCM3AP antisense RNA 1 (MCM3AP-AS1) has been reported to be an oncogenic factor in NSCLC, its detailed mechanism in NSCLC is unknown. In this study, quantitative real-time polymerase chain reaction (qRT-PCR) was performed to determine MCM3AP-AS1, microRNA (miR)-195-5p and E2F transcription factor 3 (E2F3) mRNA expressions in NSCLC tissues and cells. Western blot was utilized to determine the expression levels of E2F3, BCL2-associated X protein (Bax), B-cell lymphoma-2 (Bcl-2), E-cadherin and N-cadherin. CCK-8 and Transwell assays were conducted to examine cell proliferation, migration and invasion, respectively. Dual-luciferase reporter assay and RNA immunoprecipitation experiments were used to determine the regulatory relationships between MCM3AP-AS1 and miR-195-5p, and miR-195-5p and E2F3. We demonstrated that MCM3AP-AS1 was overexpressed in NSCLC tissues and cells, and MCM3AP-AS1 overexpression accelerated the proliferation, migration and invasion of NSCLC cells. In addition, MCM3AP-AS1 overexpression markedly up-modulated Bcl-2 expression and repressed Bax expression; MCM3AP-AS1 overexpression also significantly up-regulated N-cadherin expression and suppressed E-cadherin expression in NSCLC cells. What is more, in NSCLC cells, miR-195-5p was a target of MCM3AP-AS1, and the latter worked as a molecular sponge for miR-195-5p to regulate E2F3 expression. Collectively, MCM3AP-AS1, serving as a competitive endogenous RNA (ceRNA) to regulate miR-195-5p/E2F3 axis, promotes NSCLC progression, which is a promising therapeutic target for NSCLC.
肺癌（LC）在所有癌症相关死亡原因中位居第一，其中非小细胞肺癌（NSCLC）占肺癌病例的 85%。虽然 lncRNA MCM3AP 反义 RNA 1（MCM3AP-AS1）已被报道为 NSCLC 的致癌因子，但它在 NSCLC 中的详细机制尚不清楚。在本研究中，通过定量实时聚合酶链反应（qRT-PCR）测定 NSCLC 组织和细胞中 MCM3AP-AS1、微小 RNA（miR）-195-5p 和 E2F 转录因子 3（E2F3）mRNA 的表达。通过 Western blot 测定 E2F3、B 细胞淋巴瘤-2 相关 X 蛋白（Bax）、B 细胞淋巴瘤-2（Bcl-2）、E-钙黏蛋白和 N-钙黏蛋白的表达水平。通过 CCK-8 和 Transwell 检测分别检测细胞增殖、迁移和侵袭能力。双荧光素酶报告基因检测和 RNA 免疫沉淀实验用于确定 MCM3AP-AS1 与 miR-195-5p 以及 miR-195-5p 与 E2F3 之间的调控关系。我们证实 MCM3AP-AS1 在 NSCLC 组织和细胞中过表达，并且 MCM3AP-AS1 过表达加速了 NSCLC 细胞的增殖、迁移和侵袭。此外，MCM3AP-AS1 过表达显著上调了 Bcl-2 的表达并下调了 Bax 的表达；MCM3AP-AS1 过表达还显著上调了 NSCLC 细胞中 N-钙黏蛋白的表达并抑制了 E-钙黏蛋白的表达。更重要的是，在 NSCLC 细胞中，miR-195-5p 是 MCM3AP-AS1 的靶标，后者作为 miR-195-5p 的分子海绵来调节 E2F3 的表达。总之，MCM3AP-AS1 作为竞争性内源性 RNA（ceRNA）调节 miR-195-5p/E2F3 轴，促进 NSCLC 的进展，是 NSCLC 有前途的治疗靶点。
