长链非编码 RNA MALAT1 通过 miR-374b-5p/SRSF7 轴调控非小细胞肺癌的增殖、凋亡、迁移和侵袭。
Long non-coding RNA MALAT1 regulates proliferation, apoptosis, migration and invasion via miR-374b-5p/SRSF7 axis in non-small cell lung cancer.
机构信息
Department of Respiratory Medicine, Yantai Yuhuangding Hospital, Yantai, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Feb;24(4):1853-1862. doi: 10.26355/eurrev_202002_20363.
OBJECTIVE
Non-small cell lung cancer (NSCLC) is a common type of lung cancer. Long noncoding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) was reported to play a tumor-promoting role in NSCLC; however, the regulatory mechanism of MALAT1 in NSCLC progression remains largely unknown.
MATERIALS AND METHODS
The expression levels of MALAT1, miR-374b-5p and SRSF7 were measured by quantitative real-time polymerase chain reaction (qRT-PCR), and the protein level of SRSF7 was detected by Western blot analysis. Cell proliferation and apoptosis were determined by cell counting kit-8 (CCK-8) assay and flow cytometry, respectively. Cell migration and invasion were assessed by transwell assay. In addition, starBase3.0 software and dual-luciferase reporter assay were used to identify the correlations between miR-374b-5p and MALAT1 or SRSF7. Nude mouse xenograft assay was performed to explore the effects of MALAT1 on NSCLC in vivo.
RESULTS
We first observed that the levels of MALAT1 and SRSF7 were upregulated while miR-374b-5p was downregulated in NSCLC tissues; meanwhile, the expression level of MALAT1 was negatively correlated with miR-374b-5p and positively correlated with SRSF7. Both knockdown of MALAT1 and miR-374b-5p overexpression inhibited proliferation, migration and invasion and induced apoptosis in NSCLC cells. Then, we identified that miR-374b-5p was a target of MALAT1 and SRSF7 was the downstream of miR-374b-5p. In addition, overexpression of SRSF7 reversed the effects of MALAT1 knockdown on proliferation, apoptosis, migration and invasion in NSCLC cells. Finally, overexpression of MALAT1 suppressed NSCLC tumor growth in vivo.
CONCLUSIONS
Our results demonstrated that MALAT1 contributed to NSCLC progression through the MALAT1/miR-374b-5p/SRSF7 axis.
目的
非小细胞肺癌（NSCLC）是一种常见的肺癌类型。长链非编码 RNA（lncRNA）转移相关肺腺癌转录本 1（MALAT1）已被报道在 NSCLC 中发挥促肿瘤作用；然而，MALAT1 在 NSCLC 进展中的调节机制在很大程度上仍不清楚。
材料和方法
通过实时定量聚合酶链反应（qRT-PCR）测量 MALAT1、miR-374b-5p 和 SRSF7 的表达水平，并通过 Western blot 分析检测 SRSF7 的蛋白水平。通过细胞计数试剂盒-8（CCK-8）测定和流式细胞术分别测定细胞增殖和凋亡。通过 Transwell 测定评估细胞迁移和侵袭。此外，使用 starBase3.0 软件和双荧光素酶报告基因检测来鉴定 miR-374b-5p 与 MALAT1 或 SRSF7 之间的相关性。进行裸鼠异种移植实验以探索 MALAT1 对 NSCLC 的体内作用。
结果
我们首先观察到，在 NSCLC 组织中 MALAT1 和 SRSF7 的水平上调，而 miR-374b-5p 的水平下调；同时，MALAT1 的表达水平与 miR-374b-5p 呈负相关，与 SRSF7 呈正相关。MALAT1 的敲低和 miR-374b-5p 的过表达均抑制 NSCLC 细胞的增殖、迁移和侵袭，并诱导凋亡。然后，我们确定 miR-374b-5p 是 MALAT1 的靶基因，SRSF7 是 miR-374b-5p 的下游靶基因。此外，SRSF7 的过表达逆转了 MALAT1 敲低对 NSCLC 细胞增殖、凋亡、迁移和侵袭的影响。最后，MALAT1 的过表达抑制了体内 NSCLC 肿瘤的生长。
结论
我们的研究结果表明，MALAT1 通过 MALAT1/miR-374b-5p/SRSF7 轴促进 NSCLC 的进展。
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