Effect of m6A Methylation Modification on IncRNA ENST00000425005 in Doxorubicin Resistance and Epithelial-Mesenchymal Transition Progression in Lung Cancer Cells

Abstract

Lung cancer is the leading cause of cancer-related mortality worldwide, accounting for 18.4% of all cancer deaths. This study aims to investigate the underlying mechanism by which long non-coding RNA (lncRNA) ENST00000425005 mediates doxorubicin resistance and epithelial-mesenchymal transition (EMT) in lung cancer cells. The expression levels of ENST00000425005 in non-small cell lung cancer (NSCLC) cells were determined using quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). The protein levels of alkB homolog 5 (ALKBH5) and EMT markers (including Snail1, E-cadherin, N-cadherin, and Vimentin) were assessed using Western Blot analysis. RNA binding protein immunoprecipitation assays were conducted to detect the interaction between ENST00000425005 and ALKBH5. Cell viability was evaluated using cell counting kits assay, and cell invasion was determined by transwell assay. It was found that ENST00000425005 expression was downregulated, while ALKBH5 expression was upregulated in NSCLC cells. Additionally, ALKBH5 bound to ENST00000425005 and downregulated its expression. Overexpression of ALKBH5 reduced m6A modification and RNA levels of ENST00000425005. Moreover, co-overexpression of ENST00000425005 and ALKBH5 rescued loss of NSCLC cell viability, invasion, and doxorubicin resistance caused by overexpression of ENST00000425005. Furthermore, this co-overexpression rescued ENST00000425005-induced changes in expression of E-cadherin, Snail1, N-cadherin, and Vimentin. The reduction of m6A methylation modification on lncRNA ENST00000425005 caused by binding to ALKBH5 promoted doxorubicin resistance and EMT progression in NSCLC cells. In summary, targeting lncRNA ENST00000425005 holds promise as a therapeutic strategy for NSCLC.