Targeting EphA2 with miR-124 mediates Erlotinib resistance in K-RAS mutated pancreatic cancer
Targeting EphA2 with miR-124 mediates Erlotinib resistance in K-RAS mutated pancreatic cancer
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DOI：
10.1111/jphp.12941
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发表时间：
2019-02-01
影响因子：
3.3
通讯作者：
Han, Yong
中科院分区：
文献类型：
--
作者：
Du, Jing;He, Yuanqiao;Han, Yong
目的化疗耐药是晚期转移性胰腺癌（PC）患者生存率低的重要因素。方法将人胰腺癌细胞株Capan-1和BXPC-3与不同浓度的厄洛替尼（0、10、50和100 μ m）共同培养48 h。MTT法检测细胞相对存活率，流式细胞术检测细胞凋亡。pcDNA-EphA 2、si-EphA 2和miR-124模拟物/抑制剂的转染用于调节EphA 2和miR-124的细胞内水平。使用双荧光素酶报告基因测定探索miR-124与EphA 2的3 ' UTR之间的相互作用。与BXPC-3细胞相比，Capan-1细胞对不同浓度的厄洛替尼治疗表现出耐药性。在Capan-1细胞中EphA-2的表达显著增加，而miR-124的表达显著降低。过表达EphA 2诱导BXPC-3细胞对厄洛替尼治疗的抗性。EphA 2被鉴定为miR-124的新靶基因。miR-124过表达能够通过抑制EphA 2使Capan-1细胞对厄洛替尼的反应敏感。此外，在异种移植模型中，miR-124过表达和EphA 2抑制均使Capan-1细胞对厄洛替尼敏感。结论miR-124下调后EphA 2的表达可使K-RAS突变的PC细胞Capan-1对厄洛替尼产生耐药性。
Objectives Chemotheraputic drug resistance is a critical factor associated with the poor survival in advanced/metastatic pancreatic cancer (PC) patients. Methods Human pancreatic cell lines Capan-1 and BXPC-3 were cultured with different concentrations of erlotinib (0, 10, 50, and 100 mu m) for 48 h. The relative cell viability and apoptosis was detected using MTT assays and flow cytometry apoptosis analysis, respectively. Transfection of pcDNA-EphA2, si-EphA2 and miR-124 mimic/inhibitor was used to modulate the intracellular level of EphA2 and miR-124. The interaction between miR-124 and the 3 ' UTR of EphA2 was explored using dual luciferase reporter assay. Key findings Compared with BXPC-3 cells, Capan-1 cells showed resistance to differential concentration treatment of erlotinib. The expression of EphA-2 was significantly increased and the expression of miR-124 was significantly decreased in Capan-1 cells. Overexpressing EphA2 induced resistance of BXPC-3 cells to erlotinib treatment. And EphA2 was identified as a novel target gene for miR-124. MiR-124 overexpression was able to sensitize the response of Capan-1 cells to erlotinib through inhibiting EphA2. Furthermore, both miR-124 overexpression and EphA2 inhibition sensitized Capan-1 cells to erlotinib in xenograft model. Conclusions Our study demonstrated that EphA2 rescued by miR-124 downregulation conferred the erlotinib resistance of PC cell Capan-1 with K-RAS mutation.