基于BNIP3/BNIP3L信号通路的芹菜素诱导结直肠癌线粒体自噬的机制研究OA
Mechanism of apigenin-induced mitophagy in colorectal cancer based on BNIP3/BNIP3L signaling pathway
研究芹菜素对结直肠癌细胞线粒体自噬的影响及其基于B细胞淋巴瘤2/腺病毒E1B 19 kDa相互作用蛋白3(B-cell lymphoma 2(BCL-2)/adenovirus E1B 19 kDa protein-interacting protein 3,BNIP3)/BCL-2/腺病毒 E1B 19 kDa相互作用蛋白 3 样蛋白(BCL-2/adenovirus E1B 19 kDa protein-interacting protein 3-like,BNIP3L)信号通路的作用机制.用MTT实验研究芹菜素对人结直肠癌LoVo细胞增殖的影响;用克隆形成实验研究芹菜素对LoVo细胞克隆形成能力的影响;用透射电镜观察芹菜素对LoVo细胞线粒体自噬形态学的影响;用ATP试剂盒检测芹菜素对LoVo细胞ATP含量的影响;用JC-1试剂盒检测芹菜素对LoVo细胞线粒体膜电位的影响;用RT-qPCR检测芹菜素对LoVo细胞线粒体自噬相关mRNA表达的影响;用Western blot检测芹菜素对LoVo细胞线粒体自噬相关蛋白水平的影响.实验结果表明,与对照组比较,芹菜素能够显著抑制LoVo细胞的增殖和克隆形成,诱导LoVo细胞呈现出线粒体自噬的形态学特征.同时,芹菜素能够降低LoVo细胞ATP含量和线粒体膜电位,此作用可被线粒体自噬激动剂CCCP协同增强,又可被线粒体自噬抑制剂Mdivi-1拮抗减弱.另外,芹菜素显著升高LoVo细胞的微管相关蛋白1A/1B轻链 3(microtubule-associated protein 1A/1B-light chain 3,LC3)、BNIP3 和 BNIP3L mRNA 和蛋白表达水平(P＜0.05 或P＜0.01),但降低 P62(sequestosome 1,SQSTM1)、线粒体内膜转位酶 23 亚基(translocase of the inner mitochondrial membrane 23 kDa subunit,TIMM23)和 TOMM20mRNA 和蛋白表达水平(P＜0.05 或 P＜0.01).综上,芹菜素可通过增强结直肠癌细胞BNIP3/BNIP3L信号通路活性诱导线粒体自噬,从而发挥抗结直肠癌作用.
The study aims to investigate the effect of apigenin on the mitophagy in colorectal cancer cells and its underlying mechanism based on the B-cell lymphoma 2(BCL-2)/adenovirus E1B 19 kDa protein-interacting protein 3(BNIP3)/BCL-2/adenovirus E1B 19 kDa protein-interacting protein 3-like(BNIP3L)signaling pathway.The effect of apigenin on the prolif-eration of human colorectal cancer LoVo cells was detected by MTT assay.The efficacy of apigenin on the clonality of LoVo cells was examined by colony formation assay.The influence of apigenin on the morphology of the mitophagy in LoVo cells was observed by transmission electron microscopy.The impact of apigenin on the ATP content in LoVo cells was assessed by the ATP kit.The potency of apigenin on the mitochondrial membrane potential was evaluated by the JC-1 kit.The RT-qPCR was performed to analyze the effect of apigenin on the mitophagy-related mRNA expressions in LoVo cells.The Western blot anal-ysis was conducted to probe the influence of apigenin on the mitophagy-related protein levels in LoVo cells.Compared with the control,apigenin significantly inhibited the proliferation and colony formation of LoVo cells,and induced the morphological features of mitophagy in LoVo cells.Meanwhile,apigenin decreased the ATP content and mitochondrial membrane potential of LoVo cells,which was synergized by the activator of mitophagy named CCCP,and antagonized by the inhibitor of mitophagy named Mdivi-1.Additionally,apigenin significantly increased the mRNA and protein expression levels of microtubule-associat-ed protein 1A/1B-light chain 3(LC3),BNIP3 and BNIP3L in LoVo cells(P＜0.05 or P＜0.01),but mitigated the mRNA and protein expression levels of P62(sequestosome 1,SQSTM1),the translocase of the inner mitochondrial membrane 23 kDa subunit(TIMM23)and the translocase of the outer mitochondrial membrane 20 kDa subunit(TOMM20)in LoVo cells(P＜0.05 or P＜0.01).Taken together,apigenin can induce the mitophagy of colorectal cancer by heightening the activity of the BNIP3/BNIP3L signaling pathway,so as to exert the anti-colorectal cancer effect.
何紫雨;唐晓梦;刘晓虹;罗文雯;胡佳豪;明天琪;李煜兵;徐海波
成都中医药大学药学院西南特色中药资源国家重点实验室,成都 611137成都中医药大学药学院西南特色中药资源国家重点实验室,成都 611137成都中医药大学药学院西南特色中药资源国家重点实验室,成都 611137成都中医药大学药学院西南特色中药资源国家重点实验室,成都 611137成都中医药大学药学院西南特色中药资源国家重点实验室,成都 611137成都中医药大学药学院西南特色中药资源国家重点实验室,成都 611137成都中医药大学药学院西南特色中药资源国家重点实验室,成都 611137成都中医药大学药学院西南特色中药资源国家重点实验室,成都 611137
医药卫生
芹菜素结直肠癌线粒体自噬BNIP3BNIP3LLoVo细胞
apigenincolorectal cancermitophagyBNIP3BNIP3LLoVo cells
《天然产物研究与开发》 2026 (8)
1652-1660,1734,10
国家自然科学基金(81573813)四川省自然科学基金面上项目(2023NSFSC0653)
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