Fund programs: Hunan Province Socialized Funding Projects for Science and Technology Innovation Plan(No.2020SKC2009);Natural Science Foundation of Hunan Province(No.2024JJ7461);General Guidance Project of Hubei Provincial Health Commission(No.D202303038378);Guiding Plan Project of Science and Technology Bureau of Hengyang(No.202222035832)
Authors：Jiang Liangjun; Lu Xianzhou
Keywords：circFADS2; colorectal cancer; SW480 cells;death of iron; miR-152-3p; solute carrier family 7 member 11;
DOI：10.19405/j.cnki.issn1000-1492.2025.06.004
〔Abstract〕 Objective To investigate the effect and mechanism of silencing circFADS2 on ferroptosis in colorectal cancer (CRC) cells . Methods The human colon adenocarcinoma cell line SW480 was used as the research ob- ject. circFADS2 siRNA interference plasmid (si-circFADS2) and its negative control si-NC , miR-152-3p inhibitor and its negative control inhibitor-NC , miR-152-3p mimics and its negative control mimics NC , SLC7A11 overex- pression plasmid (oe-SLC7A11) and its negative control vector were transfected into SW480 cells . Methylthiazolyl- diphenyl-tetrazolium bromide (MTT) method was used to detect cell proliferation activity; The contents of ferrous ion (Fe2 + ) , malondialdehyde (MDA) and the activities of superoxide dismutase (SOD) and glutathione (GSH) in cells were detected by chemical method . Dichlorodihydrofluorescein diacetate ( DCFH-DA) probe was used to detect the level of reactive oxygen species ( ROS) in cells . Quantitative Real-time polymerase chain reaction (qPCR) was used to detect the expression levels of miR-152-3p and SLC7A11 mRNA in cells . Western blot was used to detect the expression levels of ferroptosis-related proteins such as SLC7A11 and GPX4 in cells . Dual lucif- erase reporter gene experiment was used to verify the sponge adsorption relationship between miR-152-3p and circFADS2 , and the targeted regulation relationship between miR-152-3p and SLC7A11 . Results Compared with blank group , the proliferation activity of si-circFADS2 group decreased , the levels of Fe2 + , MDA and ROS in- creased , and the levels of SOD and GSH decreased; At the same time , the expression level of miR-152-3p in- creased , and the protein expression levels of SLC7A11 and GPX4 decreased in cells ( all P < 0. 05) . Compared with si-circFADS2 group , the proliferation activity of si-circFADS2 + inhibitor group increased , the levels of Fe2 + , MDA and ROS in cells decreased , and the levels of SOD and GSH increased; At the same time , the expression level of miR-152-3p decreased , and the protein expression levels of SLC7A11 and GPX4 increased (all P < 0. 05) . The results of dual luciferase reporter gene experiment showed that SLC7A11 was a downstream target gene of miR- 152-3p . Conclusion Silencing circFADS2 can induce ferroptosis in CRC cells possibly by targeting the miR-152 - 3p/SLC7A11 signaling axis .