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Fig. 3 | Biomarker Research

Fig. 3

From: LncRNA APCDD1L-AS1 induces icotinib resistance by inhibition of EGFR autophagic degradation via the miR-1322/miR-1972/miR-324-3p-SIRT5 axis in lung adenocarcinoma

Fig. 3

MiR-1322/miR-1972/miR-324-3p were sponged by APCDD1L-AS1 and improved the sensitivity of LUAD cells to icotinib. a Potential miRNAs (miR-1322, miR-1972 and miR-324-3p) sponging with APCDD1L-AS1 were predicted by LncBasedatabase and transcriptome sequencing. b Sequence alignment of wild type and mutant type of APCDD1L-AS1 with miR-1322, miR-1972 and miR-324-3p potential targeting sites. c Luciferase reporter assay using miRNA mimics was applied to verify the interaction between APCDD1L-AS1 and miR-1322, miR-1972 or miR-324-3p. d RIP assay was performed using AGO2 antibody, immunoprecipitation of APCDD1L-AS1 and miR-1322, miR-1972 or miR-324-3p were determined by qRT-PCR. IgG was used as a negative control. e The expression level of miRNAs in the parental cells and their icotinib-resistant cells (PC9, PC9/IcoRL and PC9/IcoRH) was determined by qRT-PCR. f The effect of miR-1322, miR-1972 or miR-324-3p mimics on the icotinib sensitivity of PC9/IcoRL cells was determined by MTT assay. g After co-transfection with miR-1322, miR-1972 or miR-324-3p inhibitor and si-RNA APCDD1L-AS1, the viability of PC9/IcoRL cells with the treatment of icotinib for 72 h was determined by MTT assay. h-i The level of EGFR, p-EGFR (h) and PARP (i) in the icotinib-resistant cells (PC9/IcoRH and HCC827/IcoRH) with the transfection of miR-1322, miR-1972 or miR-324-3p mimics was determined by western blot. The mean ± SD of triplicate experiments were plotted, *P < 0.05, **P < 0.01, ***P < 0.001, n.s., not statistically significant

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