This study aims to explore potential novel drug resistance mechanism of NCI-H460 cells to the neddylation inhibitor MLN4924.

NCI-H460 cells were treated by MLN4924 to screen and establish drug-resistant cell lines. High throughput transcriptome sequencing was performed on the drug-resistant cell lines, MLN4924 treated 8-hour cell lines, 48-hour cell lines, and control cells. Then we screened differentially expressed genes based on sequencing results and conducted pathway enrichment analysis. The comparison among multiple groups was used by one-way ANOVA, and the pairwise comparison between two groups were performed by Dunnett-t test.

Three MLN4924-resistant cell lines were constructed. Transcriptome sequencing analysis showed that compared to the normal control group, there were 5 303, 4 186, 3 388, 3 675, and 3267 differentially expressed genes for MLN4924 treated 8-hour, 48-hour cell lines, and three drug-resistant cell lines, respectively, including genes such as ABCA9, ABCA13, CYR61, CYP39A1, etc. The RT-qPCR experiment further confirmed the significant high expression of ABCA9 and CYR61 in the transient treated cells and drug-resistant cells. The GO enrichment analysis results showed that differentially expressed genes were highly correlated with cellular biological processes such as cell cycle, apoptosis, intercellular signal transduction, and cell adhesion, as well as with cell components such as nucleus and cell membrane. KEGG pathway enrichment analysis showed that differentially expressed genes in drug-resistant cells were concentrated in MAPK signaling pathway, PI3K/Akt signaling pathway and cAMP signaling pathway.

In NCI-H460 human lung cancer cells, changes in the expression of genes such as ABCA9, CYR61, ABCC8, and CYP4F12 may enhance their resistance to MLN4924. This resistance may be related to the MAPK signaling pathway, PI3K Akt signaling pathway, and cAMP signaling pathway.