Department of Pharmacy, The First People's Hospital of Lianyungang, Lianyungang, China.
Department of Pharmacy, The First Affiliated Hospital of Kangda College of Nanjing Medical University, Lianyungang, China.
Clin Exp Pharmacol Physiol. 2022 Oct;49(10):1105-1115. doi: 10.1111/1440-1681.13692. Epub 2022 Jul 21.
Currently, a substantial amount of circular RNAs (circRNAs) are closely associated with cancer development and the occurrence of drug resistance, however, circ_0017274 in cisplatin (CDDP) resistance in gastric cancer (GC) has not been addressed. Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot were utilized for circ_0017274, microRNA-637 (miR-637) and caudal-related homeobox transcription factor 2 (CDX2) contents analysis. Analysis of IC50, proliferation, cell cycle, apoptosis, migration and invasion of GC cells using Cell Counting Kit-8 (CCK8), 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry or transwell assays. Interaction between miR-637 and circ_0017274 or CDX2 was validated under the application of luciferase reporter system, RNA immunoprecipitation (RIP) analysis, and pull-down assay. The effect of circ_0017274 on CDDP sensitivity in vivo was tapped by xenograft models. Circ_0017274 and CDX2 had higher content in CDDP-resistant GC tissues and cells, while miR-637 had lower content. CDDP resistance and development of GC cells were arrested when circ_0017274 level was reduced in vitro. MiR-637 acted as a target of circ_0017274, and miR-637 downregulation abated the phenomenon of elevated sensitivity of sh-circ_0017274 to CDDP. MiR-637 was also demonstrated to interact with CDX2 and to co-regulate CDDP sensitivity in GC cells. The xenograft models also established that circ_0017274 downregulation strengthened CDDP sensitivity and thus curtailed tumour growth in vivo. Circ_0017274 downregulation boosted CDDP sensitivity by acting on miR-637/CDX2 in CDDP-resistant GC cells.
目前,大量的环状 RNA(circRNA)与癌症的发生和耐药性的发生密切相关,然而,顺铂(CDDP)耐药性胃癌(GC)中的 circ_0017274 尚未得到解决。实时定量聚合酶链反应(RT-qPCR)和蛋白质印迹法用于 circ_0017274、微小 RNA-637(miR-637)和尾相关同源盒转录因子 2(CDX2)含量分析。使用细胞计数试剂盒-8(CCK8)、5-乙炔基-2'-脱氧尿苷(EdU)、流式细胞术或 Transwell 测定法分析 GC 细胞的 IC50、增殖、细胞周期、凋亡、迁移和侵袭。在应用荧光素酶报告系统、RNA 免疫沉淀(RIP)分析和下拉测定的情况下,验证了 miR-637 与 circ_0017274 或 CDX2 之间的相互作用。通过异种移植模型研究了 circ_0017274 对体内 CDDP 敏感性的影响。circ_0017274 和 CDX2 在 CDDP 耐药性 GC 组织和细胞中的含量较高,而 miR-637 的含量较低。体外降低 circ_0017274 水平可阻止 CDDP 耐药和 GC 细胞的发展。miR-637 是 circ_0017274 的靶标,下调 miR-637 减弱了 sh-circ_0017274 对 CDDP 的敏感性升高现象。miR-637 还被证明与 CDX2 相互作用,并共同调节 GC 细胞中的 CDDP 敏感性。异种移植模型还证实,下调 circ_0017274 可增强 CDDP 敏感性,从而在体内抑制肿瘤生长。circ_0017274 通过作用于 CDDP 耐药性 GC 细胞中的 miR-637/CDX2 来增强 CDDP 敏感性。