RBM15 silencing promotes ferroptosis by regulating the TGF-β/Smad2 pathway in lung cancer

by Selleckchem | Oct 22 2023

Objective: We assessed the function and mechanism of RNA binding motif protein 15 (RBM15) silencing in lung cancer development.

Methods: The effects of RBM15 knockdown on A549 and H1299 cells were evaluated by MTT, EdU, wound healing, and transwell assay. We then detected the functions of RBM15 silencing on lipid peroxidation, labile iron pool (LIP), ferrous iron (Fe2+ ), and ferroptosis-related genes. RNA sequencing was performed after RBM15 knockout in lung cancer cells, followed by differentially expressed genes (DEGs), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were performed. Finally, the expression of RBM15 and pathway-related genes was determined by western blot.

Results: RBM15 was highly expressed in lung cancer cells. RBM15 silencing reduced the viability, inhibited cell proliferation, invasion, and migration, and suppressed tumor growth in the xenograft mouse model. Knockout of RBM15 regulated ferroptosis-related gene expression. LIP, Fe2+ , and lipid peroxidation were distinctly increased by the knockout of RBM15. RNA-seq sequencing revealed that there are 367 up-regulated and 368 down-regulated DEGs, which were enriched in molecular functions, biological processes, and cellular components. RBM15 silencing reduced the expression of TGF-β/Smad2, and TGF-β activator (SRI-011381) reversed the inhibitory effect of RBM15 silencing on tumor cell growth.

Conclusion: We demonstrated that RBM15 silencing promoted ferroptosis in lung cancer cells by TGF-β/Smad2 pathway, thereby inhibiting lung cancer cell growth, which may provide new light for lung cancer treatment.

Comments:

Summary: The objective of the study was to investigate the function and mechanism of RNA binding motif protein 15 (RBM15) silencing in lung cancer development. The researchers used various experimental methods to assess the effects of RBM15 knockdown in lung cancer cells (A549 and H1299) including MTT assay, EdU assay, wound healing assay, and transwell assay. They also examined the impact of RBM15 silencing on lipid peroxidation, labile iron pool (LIP), ferrous iron (Fe2+), and ferroptosis-related genes. RNA sequencing was performed after RBM15 knockout, followed by analysis of differentially expressed genes (DEGs), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG). The expression of RBM15 and pathway-related genes was determined by western blot.

The results showed that RBM15 was highly expressed in lung cancer cells. Silencing RBM15 reduced cell viability, inhibited proliferation, invasion, and migration, and suppressed tumor growth in a mouse xenograft model. Knockout of RBM15 affected the expression of ferroptosis-related genes and increased levels of LIP, Fe2+, and lipid peroxidation. RNA sequencing revealed a total of 367 up-regulated and 368 down-regulated DEGs, which were enriched in various molecular functions, biological processes, and cellular components. RBM15 silencing also reduced the expression of TGF-β/Smad2, and the addition of a TGF-β activator reversed the inhibitory effect of RBM15 silencing on tumor cell growth.

In conclusion, the study demonstrated that RBM15 silencing promoted ferroptosis in lung cancer cells through the TGF-β/Smad2 pathway, leading to the inhibition of lung cancer cell growth. These findings provide new insights for potential treatments in lung cancer.