Circular RNA AKT3 upregulates PIK3R1 to enhance cisplatin resistance in gastric cancer via miR-198 suppression
Circular RNA AKT3 upregulates PIK3R1 to enhance cisplatin resistance in gastric cancer via miR-198 suppression
复制标题
环状RNA AKT3通过抑制miR-198上调PIK3R1以增强胃癌的顺铂耐药性
DOI：
10.1186/s12943-019-0969-3
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发表时间：
2019-03-30
期刊：
影响因子：
37.3
通讯作者：
Xu, Zekuan
中科院分区：
文献类型：
--
作者：
Huang, Xiaoxu;Li, Zheng;Xu, Zekuan
背景顺铂（CDDP）治疗是胃癌（GC）患者最主要的化疗策略之一。更好地了解CDDP耐药机制可以大大提高GC患者的治疗效果。环状RNA（circRNA）是一类非编码RNA，其功能与癌症的发病机制相关，但GC的CDDP耐药性仍不清楚。方法通过RNA测序鉴定circAKT3（hsa_circ_0000199，源自AKT3基因第8、9、10和11号外显子的circRNA），并通过定量逆转录PCR进行验证。在体外和体内评估了 circAKT3 在 GC CDDP 抗性中的作用。采用荧光素酶报告基因测定、生物素偶联 RNA Pull-down 和荧光原位杂交 (FISH) 来评估 circAKT3 和 miR-198 之间的相互作用。通过蛋白质印迹、细胞毒性试验、克隆形成试验和流式细胞术进行功能实验。结果CDDP 耐药的 GC 组织和细胞中 circAKT3 的表达高于 CDDP 敏感样本。接受 CDDP 治疗的 GC 患者中 circAKT3 的上调与侵袭性特征显着相关，并且是无病生存 (DFS) 的独立危险因素。我们的数据表明，circAKT3 在体内和体外促进 DNA 损伤修复并抑制 GC 细胞凋亡。从机制上，我们验证了circAKT3可以通过海绵miR-198促进PIK3R1的表达。结论circAKT3在GC对CDDP的抵抗中发挥重要作用。因此，我们的结果强调了 circAKT3 作为接受 CDDP 治疗的 GC 患者的治疗靶点的潜力。
BackgroundCisplatin (CDDP) treatment is one of the most predominant chemotherapeutic strategies for patients with gastric cancer (GC). A better understanding of the mechanisms of CDDP resistance can greatly improve therapeutic efficacy in patients with GC. Circular RNAs (circRNAs) are a class of noncoding RNAs whose functions are related to the pathogenesis of cancer, but, in CDDP resistance of GC remains unknown.MethodscircAKT3 (hsa_circ_0000199, a circRNA originating from exons 8, 9, 10, and 11 of the AKT3 gene) was identified by RNA sequencing and verified by quantitative reverse transcription PCR. The role of circAKT3 in CDDP resistance in GC was assessed both in vitro and in vivo. Luciferase reporter assay, biotin-coupled RNA pull-down and fluorescence in situ hybridization (FISH) were conducted to evaluate the interaction between circAKT3 and miR-198. Functional experiments were measured by western blotting, a cytotoxicity assay, clonogenic assay and flow cytometry.ResultsThe expression of circAKT3 was higher in CDDP-resistant GC tissues and cells than in CDDP-sensitive samples. The upregulation of circAKT3 in GC patients receiving CDDP therapy was significantly associated with aggressive characteristics and was an independent risk factor for disease-free survival (DFS). Our data indicated that circAKT3 promotes DNA damage repair and inhibits the apoptosis of GC cells in vivo and in vitro. Mechanistically, we verified that circAKT3 could promote PIK3R1 expression by sponging miR-198.ConclusionscircAKT3 plays an important role in the resistance of GC to CDDP. Thus, our results highlight the potential of circAKT3 as a therapeutic target for GC patients receiving CDDP therapy.