Abstract
The Molecular Mechanism of Circular RNA Molecule CircSETD3 in Gefitinib Acquired Resistance in Non-Small Cell Lung Cancer
Department of Respiratory Medicine, The First Affiliated Hospital of Hebei North University, 1Department of Anatomy, School of Basic Medicine, Hebei North University, Zhangjiakou, Hebei 075000, China
Correspondence Address:
Y. Huo, Department of Anatomy, School of Basic Medicine, Hebei North University, Zhangjiakou, Hebei 075000, China, E-mail: yaocangdang5@163.com
To explore the molecular pathway of the circular RNA molecule in gefitinib acquired resistance in nonsmall cell lung cancer and to provide a new therapeutic direction for clinical treatment of non-small cell lung cancer. The gefitinib-sensitive non-small cell lung cancer cell strain H292_S was selected to construct gefitinib acquired resistant cell model H292_R. Also, the overexpression group and vector control group of H292_S transfected circSETD3; the knockdown plasmid group and vector control group of H292_R transfected circSETD3 were prepared. The degree of cell response to gefitinib was measured by Western blotting and cell counting kit-8 method. The RNA sequencing method was used to screen differentially expressed circular RNA in cells and real-time fluorescent quantitative polymerase chain reaction was used to verify the sequencing results; the dual-luciferase report was used to screen and verify the relationship between circSETD3 and microRNA-520h. After the drug-resistant H292_R cell strain was processed by gefitinib, the expression of phosphorylated epidermal growth factor receptor, phosphatidylinositol 3-kinase and protein kinase B decreased significantly. The expressions of three circular RNAs in has_circ_0000567, has_circ_0000655 and has_circ_0006867, were all up-regulated in drug-resistant strains, and the differences were statistically significant (p<0.05). The overexpression group of circSETD3 had a significant effect on the growth and reproduction ability of cells (p<0.05); the wild type group of the dual-luciferase report plasmid was transfected into 293T cells together with microRNA-520h mimics and the fluorescence intensity decreased significantly (p<0.05). In addition, if the luciferase reporter plasmid mutant group (mutant) was transfected into 293T cells together with microRNA-520h mimics, the fluorescence intensity would not change significantly. circSETD3 could be directly bound to microRNA-520h.
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