Department of Thoracic Surgery, Southwest Hospital, Chongqing Ninth People's Hospital, Chongqing, China.
Department of Cardiothoracic Surgery, Chongqing Hospital of Traditional Chinese Medicine, Chongqing, China
Ann Clin Lab Sci. 2023 Jan;53(1):64-75.
Non-small cell lung cancer (NSCLC) is recognized as one of the primary causes of global cancer-related mortality. Long noncoding RNAs (lncRNAs) participate in NSCLC cell progression. This study probed the potential mechanism of lncRNA small nucleolar RNA host gene 12 (SNHG12) in cisplatin (DDP)-resistance in NSCLC cells.
The intracellular expressions of SNHG12, miR-525-5p, and XIAP were examined via reverse-transcription quantitative polymerase chain reaction (RT-qPCR). Afterwards, small interfering RNAs (siRNAs) of SNHG12, microRNA (miR)-525-5p inhibitor, and X-linked inhibitor of apoptosis (XIAP) pcDNA3.1 were transfected into NSCLC cells. Subsequently, changes in half-maximal (50%) inhibitory concentration (IC) of NSCLC cells to DDP were detected through the cell counting kit-8 (CCK-8) method. NSCLC proliferative ability and apoptosis rate were determined with the help of colony formation and flow cytometry assays. The subcellular localization of SNHG12 was analyzed by nuclear/cytosol fractionation assay and binding relationships between miR-525-5p and SNHG12 or XIAP were analyzed via dual-luciferase reporter gene assay. Furthermore, rescue experiments were designed to detect the effects of miR-525-5p and XIAP on NSCLC sensitivity to DDP.
SNHG12 and XIAP were up-regulated in NSCLC cells while miR-525-5p was down-regulated. After DDP treatment and SNHG12 repression, NSCLC proliferative ability was decreased whereas apoptosis rate was increased, and NSCLC sensitivity to DDP was enhanced. Mechanically, SNHG12 repressed miR-525-5p expression, and miR-525-5p could targeted inhibit XIAP transcription level. miR-525-5p repression or XIAP overexpression reduced NSCLC sensitivity to DDP.
SNHG12 was overexpressed in NSCLC cells and promoted XIAP transcription by repressing miR-525-5p expression, enhancing DDP-resistance in NSCLC cells.
非小细胞肺癌（NSCLC）是全球癌症相关死亡的主要原因之一。长链非编码 RNA（lncRNA）参与 NSCLC 细胞的进展。本研究探讨了 lncRNA 小核仁 RNA 宿主基因 12（SNHG12）在 NSCLC 细胞顺铂（DDP）耐药中的潜在机制。
采用逆转录定量聚合酶链反应（RT-qPCR）检测 SNHG12、miR-525-5p 和 XIAP 的细胞内表达。然后，将 SNHG12 的小干扰 RNA（siRNA）、miR-525-5p 抑制剂和 X 连锁凋亡抑制剂（XIAP）pcDNA3.1 转染到 NSCLC 细胞中。随后，通过细胞计数试剂盒-8（CCK-8）法检测 NSCLC 细胞对 DDP 的半数最大抑制浓度（IC）的变化。通过集落形成和流式细胞术检测 NSCLC 增殖能力和凋亡率。通过核/浆分离测定分析 SNHG12 的亚细胞定位，并通过双荧光素酶报告基因测定分析 miR-525-5p 与 SNHG12 或 XIAP 的结合关系。此外，还设计了挽救实验来检测 miR-525-5p 和 XIAP 对 NSCLC 对 DDP 敏感性的影响。
SNHG12 和 XIAP 在 NSCLC 细胞中上调，而 miR-525-5p 下调。在 DDP 处理和 SNHG12 抑制后，NSCLC 增殖能力降低，凋亡率增加，对 DDP 的敏感性增强。机制上，SNHG12 抑制 miR-525-5p 的表达，miR-525-5p 可以靶向抑制 XIAP 的转录水平。miR-525-5p 抑制或 XIAP 过表达降低了 NSCLC 对 DDP 的敏感性。
SNHG12 在 NSCLC 细胞中过度表达，并通过抑制 miR-525-5p 的表达促进 XIAP 的转录，增强 NSCLC 细胞对 DDP 的耐药性。
