长链非编码 RNA SNHG14 通过 miR-133a/HOXB13 通路调节非小细胞肺癌细胞对顺铂的耐药性。
LncRNA SNHG14 regulates the DDP-resistance of non-small cell lung cancer cell through miR-133a/HOXB13 pathway.
机构信息
Thoracic Medicine Department 2, The Affiliated Cancer Hospital of Xiangya School of Medicine and Hunan Cancer Hospital, Central South University, No. 283, Tongzipo Road, Yuelu District, Changsha, 410013, Hunan Province, PR China.
Respiratory Medicine Department 2, Hunan Children's Hospital, Changsha, 410007, Hunan Province, PR China.
出版信息
BMC Pulm Med. 2020 Oct 15;20(1):266. doi: 10.1186/s12890-020-01276-7.
BACKGROUND
Recently, long non-coding RNAs (lncRNAs) have been reported to be involved in regulating chemo-resistance of NSCLC, however, the role of lncRNA SNHG14 in the DDP-resistance of NSCLC remains unexplored.
METHODS
Relative expression of SNHG14, HOXB13 and miR-133a in DDP-resistant A549 (A549/DDP) cell and its parental cell A549 were measured using qRT-PCR. Cell proliferation viability of indicated A549/DDP cell was estimated via CCK-8 and colony formation experiments. Cell cycle and apoptosis were analyzed through flow cytometry. Expression of apoptosis-related protein and HOXB13 were detected via western blot. The interaction among SNHG14, HOXB13 and miR-133a was predicted by bioinformatics and validated by dual-luciferase reporter assay.
RESULTS
LncRNA SNHG14 and HOXB13 were upregulated while miR-133a was downregulated in A549/DDP cell line compared to A549 cell line. SNHG14 knockdown or miR-133a overexpression was demonstrated to increase the DDP-sensitivity of A549/DDP cells. SNHG14 was revealed to compete with HOXB13 for miR-133a binding in A549/DDP cells. Inhibition of miR-133a in A549 cells could reverse the promotive effects of SNHG14 knockdown on DDP-sensitivity, as well as the inhibitory effects on HOXB13 expression. HOXB13 overexpression was revealed to abolish the enhanced effects of miR-133a on the sensitivity of A549/DDP cell to DDP.
CONCLUSION
Our findings demonstrated that SNHG14 was involved in the development of DDP-resistance of A549/DDP cells through miR-133a/HOXB13 axis, which may present a path to novel therapeutic stratagems for DDP resistance of NSCLC.
背景
最近，长链非编码 RNA（lncRNA）已被报道参与调节 NSCLC 的化疗耐药性，然而，lncRNA SNHG14 在 NSCLC 的 DDP 耐药中的作用仍未被探索。
方法
采用 qRT-PCR 检测 DDP 耐药的 A549（A549/DDP）细胞及其亲本细胞 A549 中 SNHG14、HOXB13 和 miR-133a 的相对表达水平。通过 CCK-8 和集落形成实验评估指示的 A549/DDP 细胞的细胞增殖活力。通过流式细胞术分析细胞周期和凋亡。通过 Western blot 检测凋亡相关蛋白和 HOXB13 的表达。通过生物信息学预测 SNHG14、HOXB13 和 miR-133a 之间的相互作用，并通过双荧光素酶报告实验验证。
结果
与 A549 细胞系相比，A549/DDP 细胞系中 lncRNA SNHG14 和 HOXB13 上调，而 miR-133a 下调。SNHG14 敲低或 miR-133a 过表达可增加 A549/DDP 细胞对 DDP 的敏感性。在 A549/DDP 细胞中，SNHG14 被发现与 HOXB13 竞争与 miR-133a 的结合。在 A549 细胞中抑制 miR-133a 可逆转 SNHG14 敲低对 DDP 敏感性的促进作用，以及对 HOXB13 表达的抑制作用。过表达 HOXB13 可消除 miR-133a 对 A549/DDP 细胞对 DDP 敏感性的增强作用。
结论
我们的研究结果表明，SNHG14 通过 miR-133a/HOXB13 轴参与 A549/DDP 细胞 DDP 耐药的发展，这可能为 NSCLC 的 DDP 耐药提供新的治疗策略。
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