| 黎文涛,伍小琼,冯志鹏,王正根.过表达miR-199-3p/5p诱导胰腺癌细胞BxPC-3、Panc-1铁死亡的作用机制.[J].中南医学科学杂志.,2023,(3):344-348. |
| 过表达miR-199-3p/5p诱导胰腺癌细胞BxPC-3、Panc-1铁死亡的作用机制 |
| Mechanism of ferroptosis induced by overexpression of miR-199-3p/5p in pancreatic adenocarcinoma cells BxPC-3 and Panc-1 |
| 投稿时间：2022-11-04 修订日期：2023-03-23 |
| DOI：10.15972/j.cnki.43-1509/r.2023.03.007 |
| 中文关键词: miR-199-3p miR-199-5p 胰腺癌 铁死亡 [ |
| 英文关键词:miR-199-3p miR-199-5p pancreatic adenocarcinoma ferroptosis |
| 基金项目:湖南省卫生健康委科研计划项目(202203103861) |
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| 中文摘要: |
| 目的探讨过表达miR-199-3p/5p诱导胰腺癌细胞BxPC-3和Panc-1铁死亡的作用机制。 方法瞬时转染miR-199-3p/5p模拟物和抑制物至BxPC-3、Panc-1细胞中。qRT-PCR检测细胞中miR-199-3p/5p表达水平；Western blotting检测SCL7A11、BTRC、TFRC蛋白水平。谷胱甘肽(GSH)、Fe2＋、脂质过氧化物(LPO)试剂盒检测细胞GSH、Fe2＋、LPO水平。免疫共沉淀检测BTRC、TFRC泛素化水平。双荧光素酶报告实验检测miR-199与靶基因的相互作用。 结果转染miR-199-3p/5p模拟物后,胰腺癌BxPC-3、Panc-1细胞株中miR-199-3p/5p、TFRC蛋白水平显著上调；SLC7A11、BTRC蛋白水平显著降低；LPO、Fe2＋含量升高,GSH含量降低。过表达BTRC能够显著降低TFRC蛋白水平。miR-199-3p靶向结合SLC7A11,miR-199-5p靶向结合BTRC。 结论过表达miR-199-3p/5p诱导胰腺癌细胞BxPC-3和PANC-1铁死亡,可能与靶向抑制BTRC、SLC7A11促进LPO累积有关。 |
| 英文摘要: |
| AimTo investigate the mechanism of ferroptosis induced by overexpression of miR-199-3p/5p in pancreatic adenocarcinoma cells BxPC-3 and Panc-1. MethodsTransient transfection of miR-199-3p/5p mimic and inhibitors into BxPC-3 and Panc-1 cells. qRT-PCR was detected the expression levels of miR-199-3p/5p in cells; Western blotting were used to detect SCL7A11, BTRC, TFRC protein expression levels. Glutathione (GSH), Fe2＋ and lipid peroxide (LPO) kits were used to detect the levels of GSH, Fe2＋ and LPO. The regulation of ubiquitination of BTRC and TFRC was detected by immunoprecipitation. The interaction between miR-199 and target genes was detected by dual luciferase reporting assay. ResultsAfter transfection of miR-199-3p/5p mimic in pancreatic adenocarcinoma cells BxPC-3 and Panc-1, the levels of miR-199-3p/5p and TFRC protein were significantly up-regulated, and the expression levels of SLC7A11 and BTRC proteins decreased significantly; The level of GSH decreased, and the levels of Fe2＋ and LPO increased significantly. Overexpression of BTRC can significantly reduce TFRC protein level. miR-199-3p binds to SLC7A11, and miR-199-5p binds to BTRC. ConclusionOverexpression of miR-199-3p/5p induces ferroptosis in pancreatic adenocarcinoma cells BxPC-3 and Panc-1, which may be related to the targeted inhibition of BTRC and SLC7A11 to promote LPO accumulation. |
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