AGAP2-AS1 affects TNM staging and prognosis of lung cancer patients by acting on SLC7A11 mRNA stability and ferroptosis*

Abstract

Abstract Objective The initiation and progression of lung carcinomas are critically regulated by long non-coding RNAs (lncRNAs). However, the role of lncRNAs in the pathways causing lung cancer remains unknown. Methods Cell morphology was regularly observed using an inverted phase-contrast microscope. Cell viability was assessed using CCK-8 according to the manufacturer’s instructions. Total RNA was retrotranscribed from each specimen using the RNAiso Plus Kit. The RT-PCR data were calculated using the Ct approach for comparison. Flow cytometric analyses were prepared by Click-iT™ Plus TUNEL Assay for In Situ apoptosis detection, with Alexa Fluor™ 594 dye, as instructed. RNA immunoprecipitation assays were used to determine RNA concentration. Results Activated natural killer cells repeat and PH domain-containing protein 2 antisense RNA 1 (AGAP2-AS1) levels in cancerous tissues were significantly correlated with cancerous tumor node metastasis (TNM) stage, with cancerous AGAP2-AS1 levels being higher in cancerous tissues than healthy tissues. Patients withelevated AGAP2-AS1 levels had considerably worse outcomes than those with reduced AGAP2-AS1 levels, regardless of the progression-free or overall survival. Functionally, AGAP2-AS1 downregulation represseslung cancer cell growth. AGAP2-AS1 elimination induces erastin-mediated ferroptosis in lung cancer cells.However, the ferritin inhibitor FERSINT-1 negated this result, whereas ERASTIN induced lung cancer cellmortality. After AGAP2-AS1 silencing, erastin-treated lung cancer cells showed a remarkable decrease inGSH levels. These results indicated that AGAP2-AS1 enhanced the stabilization of SLC7A11 mRNA via Recombinant Insulin Like Growth Factor Binding Protein 2(IGF BP2). Patients with elevated AGAP2-AS1 had considerably worse outcomes. Down-regulating AGAP2-AS1 was able to repress lung cancer cell growth and induce greater Erastin-mediated ferroptosis. Lungcancer cells treated with Erastin exhibited a remarkable decrease inglutathione (GSH) levels. The mechanical findingsindicated that AGAP2-AS1 enhanced the stabilization of SLC7A11 mRNA via the IGF2BP2. Conclusion We identified a novel effect of AGAP2-AS1 on TNM staging and the prognosis of patientswith lungcancer by modulating SLC7A11 mRNA stability and ferroptosis.