Feng Jing, Li Yaling, He Fen, Zhang Fuwei
Department of Thoracic Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou City, Guangdong Province, China.
Environ Toxicol. 2023 Mar;38(4):950-961. doi: 10.1002/tox.23741. Epub 2023 Jan 30.
We assessed the function and mechanism of RNA binding motif protein 15 (RBM15) silencing in lung cancer development.
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 (Fe ), 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.
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, Fe , 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.
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.
我们评估了RNA结合基序蛋白15（RBM15）沉默在肺癌发生发展中的作用及机制。
通过MTT、EdU、伤口愈合和Transwell实验评估RBM15敲低对A549和H1299细胞的影响。然后我们检测了RBM15沉默对脂质过氧化、不稳定铁池（LIP）、亚铁离子（Fe²⁺）和铁死亡相关基因的作用。在肺癌细胞中敲除RBM15后进行RNA测序，随后进行差异表达基因（DEG）、基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析。最后，通过蛋白质免疫印迹法测定RBM15和通路相关基因的表达。
RBM15在肺癌细胞中高表达。RBM15沉默降低了细胞活力，抑制了细胞增殖、侵袭和迁移，并抑制了异种移植小鼠模型中的肿瘤生长。敲除RBM15调节铁死亡相关基因的表达。敲除RBM15后，LIP、Fe²⁺和脂质过氧化明显增加。RNA测序显示有367个上调和368个下调的差异表达基因，这些基因在分子功能、生物学过程和细胞成分中富集。RBM15沉默降低了TGF-β/Smad2的表达，而TGF-β激活剂（SRI-011381）逆转了RBM15沉默对肿瘤细胞生长的抑制作用。
我们证明RBM15沉默通过TGF-β/Smad2通路促进肺癌细胞铁死亡，从而抑制肺癌细胞生长，这可能为肺癌治疗提供新的思路。
