Yan Jingyi, Chen Xiaolei, Lin Ji, Sun Xuecheng, Chen Wei
Department of Gastroenterology and General Surgery, The Quzhou Affiliated Hospital of Wenzhou Medical University, Quzhou People's Hospital, Quzhou, Zhejiang, China.
Department of Gastroenterology and General Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.
Neoplasma. 2023 Aug;70(4):526-533. doi: 10.4149/neo_2023_230523N272.
The aim of this study was to explore the role and mechanism of long non-coding RNA (lncRNA) HIF1A antisense RNA 2 (HIF1A-AS2) in regulating imatinib (IM) resistance in gastrointestinal stromal tumor (GIST) cells under hypoxia. The expression of HIF1A-AS2 was silenced by siRNA in GIST cells. Cytotoxicity, apoptosis, and autophagy were evaluated under normoxic and hypoxic conditions. The expression levels of HIF1A-AS2, HIF1A, apoptosis-associated genes, and autophagy-associated genes were determined by qRT-PCR analysis and western blot. We found that lncRNA HIF1A-AS2 was highly expressed in GIST tissues and cells. Knockdown of HIF1A-AS2 increased the sensitivity of GIST cells to IM and increased apoptosis. Moreover, a hypoxic environment decreased the sensitivity of GIST cells to IM, and the knockdown of HIF1A-AS2 reversed this effect. Mechanistically, the knockdown of HIF1A-AS2 inhibited IM-mediated autophagy. Finally, HIF1A was found to positively regulate HIF1A-AS2 under hypoxic conditions. Collectively, these data demonstrate that hypoxia-induced HIF1A-AS2 promotes IM resistance in GIST cells by regulating autophagy.
本研究旨在探讨长链非编码RNA（lncRNA）HIF1A反义RNA 2（HIF1A-AS2）在缺氧条件下调节胃肠道间质瘤（GIST）细胞对伊马替尼（IM）耐药中的作用及机制。通过小干扰RNA（siRNA）使GIST细胞中HIF1A-AS2的表达沉默。在常氧和缺氧条件下评估细胞毒性、细胞凋亡和自噬情况。采用实时定量聚合酶链反应（qRT-PCR）分析和蛋白质免疫印迹法检测HIF1A-AS2、HIF1A、凋亡相关基因和自噬相关基因的表达水平。我们发现lncRNA HIF1A-AS2在GIST组织和细胞中高表达。敲低HIF1A-AS2可增加GIST细胞对IM的敏感性并促进细胞凋亡。此外，缺氧环境会降低GIST细胞对IM的敏感性，而敲低HIF1A-AS2可逆转这一效应。机制上，敲低HIF1A-AS2可抑制IM介导的自噬。最后，发现在缺氧条件下HIF1A可正向调节HIF1A-AS2。综上所述，这些数据表明缺氧诱导的HIF1A-AS2通过调节自噬促进GIST细胞对IM的耐药。
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