解剖科学进展

目的 探究circRNA-ABCB10介导ZEB1对非小细胞肺癌(NSCLC)进展和其对表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKI)耐药性的影响及机制。方法 采用慢病毒转染建立稳定敲减circRNA-ABCB10以及在敲减circRNA-ABCB10的基础上过表达ZEB1的HCC827细胞，RT-qPCR和Western blot验证转染效率；CCK-8、流式细胞术和Transwell依次用于检测细胞增殖能力、凋亡水平以及迁移和侵袭能力；采用浓度递增法建立奥希替尼耐药的HCC827-OR细胞和吉非替尼耐药的HCC827-GR细胞，CCK-8检测不同浓度药物处理对细胞活力的影响；将转染后细胞皮下注射于裸鼠腋下并检测肿瘤体积和质量。生物信息学分析miR-876-5p与circRNA-ABCB10和ZEB1 mRNA的潜在结合位点并通过双荧光素酶报告基因实验进行验证。结果 过表达ZEB1逆转敲减circRNA-ABCB10对HCC827细胞体外增殖、迁移和侵袭以及体内移植瘤生长的抑制作用，并逆转敲减circRNA-ABCB10对HCC827细胞体外凋亡的促进作用。过表达ZEB1还逆转了敲减circRNA-ABCB10对奥希替尼和吉非替尼耐药的HCC827细胞对相应药物的耐药性的抑制作用。双荧光素酶报告基因实验证实miR-876-5p与circRNA-ABCB10和ZEB1 mRNA靶向结合。结论 CircRNA-ABCB10通过上调ZEB1促进NSCLC进展以及其对EGFR-TKI的耐药性，其机制可能与结合miR-876-5p有关。

Objective To explore the effect and mechanism of circRNA-ABCB10 on the progression of non-small ZEB1 lung cancer(NSCLC) and its resistance to epidermal growth factor receptor tyrosine kinase inhibitor(EGFR-TKI).Methods HCC827 cells with stable knockdown of circRNA-ABCB10 and overexpression of ZEB1 on the basis of knockdown of circRNA-ABCB10 were established by lentivirus transfection, and RT-qPCR and Western blot were used to verify the transfection efficiency. CCK-8, flow cytometry and Transwell assay were used to detect proliferation, apoptosis, migration and invasion of cells in turn. Oxitinib-resistant HCC827-OR cells and gefitinib-resistant HCC827-GR cells were established by increasing concentration method, and CCK-8 was used to detect the effects of different concentrations of drugs on cell viability. The transfected cells were injected subcutaneously into the armpit of nude mice and the tumor volume and quality were detected. Bioinformatics analysis of the potential binding sites of miR-876-5p with circRNA-ABCB10 and ZEB1 mRNA and verification by double luciferase reporter gene experiment.Results Over-expression of ZEB1 reversed the inhibitory effect of knockdown circRNA-ABCB10 on proliferation, migration and invasion of HCC827 cells in vitro and the growth of transplanted tumor in vivo, and reversed the promotion effect of knockdown circRNA-ABCB10 on apoptosis of HCC827 cells in vitro. Over-expression of ZEB1 also reversed the inhibitory effect of knockdown circRNA-ABCB10 on the drug resistance of HCC827 cells resistant to oxitinib and gefitinib. The experiment of double luciferase reporter gene confirmed that miR-876-5p binds to circRNA-ABCB10 and ZEB1 mRNA.Conclusion CircRNA-ABCB10 promotes the progression of NSCLC and its resistance to EGFR-TKI by up-regulating ZEB1, and its mechanism may be related to the binding of miR-876-5p.