-mediated m6A mRNA modification of suppresses cisplatin resistance in non-small cell lung cancer.

BACKGROUND

The methylation of adenosines at the N6 position (N6-methyladenosine; m6A) is one of the most conserved internal RNA modifications. m6A can modulate the expression of oncogenes or tumor suppressor genes, as well as m6A levels and the expression and activity of m6A enzymes, thus influencing tumor progression and therapeutic response. This study investigates the role of -mediated m6A messenger RNA (mRNA) modification of in controlling cisplatin resistance in non-small cell lung cancer (NSCLC).

METHODS

The expression of the m6A reader protein was detected in an NSCLC cisplatin-resistant cell line (A549/DDP) using real-time fluorescence quantitative polymerase chain reaction (qPCR). overexpression plasmids were constructed and transfected into A549/DDP and A549 cells respectively. We performed qPCR and western blot (WB) to detect changes in and Id3 expression, and the effects of overexpression on proliferation, apoptosis, invasion, and migration of drug-resistant cells were assessed by cell counting kit-8 (CCK-8), flow cytometry, and transwell and scratch assays. The m6A modification of Id3 by was clarified by m6A-immunoprecipitation-PCR (m6A-IP-PCR) assay.

RESULTS

The CLIPdb online database predicted that might bind to Id3. The results of qPCR showed that was downregulated in the NSCLC cisplatin-resistant cell line A549/DDP compared to the cisplatin-sensitive cell line A549. Overexpression of increased the expression of , and the methylation inhibitor 3-deazaadenosine abrogated the regulatory effect of on . overexpression significantly inhibited A549/DDP cell proliferation, migration, and invasion, and promoted apoptosis by synergistically promoting the effects of . m6A-IP-PCR analysis revealed that could inhibit the m6A level of mRNA.

CONCLUSIONS

To regulate the activity of , requires modifications to m6A, which ultimately inhibit cisplatin resistance in NSCLC.