摘要
目的探讨miR-1291/雌激素相关受体α(estrogen-related receptorα,ERRα)/肉碱棕榈酰转移酶1C(carnitine palmitoyltransferase 1C,CPT1C)轴在卵巢癌(ovarian cancer,OC)细胞增殖代谢表型中的作用以及绞股蓝皂苷L(gypenoside L,Gyp-L)对OC细胞miR-1291/ERRα/CPT1C轴的调控机制。方法OVCAR3细胞予以Gyp-L干预,分为对照组与Gyp-L组。CCK8法检测Gyp-L对OVCAR3细胞增殖的影响;细胞集落形成实验与划痕实验反映OVCAR3细胞的增殖与迁移能力;PANDORA-seq小RNA测序检测两组细胞差异表达的miRNA;通过试剂盒检测ROS与ATP含量、葡萄糖摄取能力以及RT-qPCR检测线粒体发生相关基因表达[核呼吸因子1(nuclear respiratory factor 1,NRF1)、过氧化物酶体增殖物激活受体γ辅助激活因子(peroxisome proliferator-activated receptor gamma coactivator,PGC-1A)、细胞色素B-245α链(cytochrome B-245 alpha chain,CYBA)、线粒体转录因子A(mitochondrial transcription factor A,TFAM)],以上实验共同反映Gyp-L对OC细胞代谢的影响;RT-qPCR检测miR-1291、ERRα及CPT1C基因表达;Western Blot检测ERRα及CPT1C蛋白表达。取8例OC患者的肿瘤组织与癌旁组织,RT-qPCR检测miR-1291、ERRα及CPT1C基因表达;Western Blot检测ERRα及CPT1C蛋白表达。结果与对照组比较,Gyp-L能显著抑制OVCAR3细胞增殖与迁移(P<0.05);Gyp-L有效干扰OVCAR3细胞中44个miRNA的表达(P<0.05);Gyp-L干预后,OVCAR3细胞miR-1291水平升高、ERRα及CPT1C表达水平降低(P<0.05);Gyp-L可升高OVCAR3细胞ROS水平并降低ATP含量、线粒体发生相关基因(NRF1、PGC-1A、CYBA、TFAM)表达降低(P<0.05)、葡萄糖摄取能力减弱(P<0.05)。在临床OC组织中发现miR-1291/ERRα/CPT1C轴存在变化(P<0.05)。结论miR-1291/ERRα/CPT1C信号通路参与OC发生发展。Gyp-L通过高表达miR-1291靶向ERRα/CPT1C信号轴抑制OVCAR3细胞增殖和代谢,从而控制OC发生发展。
Objective To investigate the role of miR-1291/estrogen-related receptorα(ERRα)/carnitine palmitoyltransferase 1C(CPT1C)axis in the proliferation and metabolic phenotype of ovarian cancer(OC)cells and the regulatory mechanism of gypenoside L(Gyp-L)on miR-1291/ERRα/CPT1C axis in OC cells.Methods OVCAR3 cells were treated with Gyp-L and divided into control group and Gyp-L group.CCK8 assay was used to detect the effect of Gyp-L on the proliferation of OVCAR3 cells.The proliferation and migration ability of OVCAR3 cells were evaluated by cell colony formation assay and wound healing assay.PANDORA-seq small RNA sequencing was used to detect the differentially expressed miRNAs between the two groups.The contents of ROS and ATP and glucose uptake ability were detected by kit.RT-qPCR was used to detect the expression of mitochondria-related genes[nuclear respiratory factor 1(NRF1),peroxisome proliferator-activated receptor gamma coactivator(PGC-1A),cytochrome B-245 alpha chain(CYBA)and mitochondrial transcription factor A(TFAM)].The above experiments reflected the effect of Gyp-L on OC cell metabolism.RT-qPCR was used to detect the expressions of miR-1291,ERRαand CPT1C genes.The protein expressions of ERRαand CPT1C were detected by Western Blot.Tumor tissues and adjacent tissues of 8 OC patients were collected,and RT-qPCR was used to detect the expressions of miR-1291,ERRαand CPT1C genes.The protein expressions of ERRαand CPT1C were detected by Western Blot.Results Gyp-L significantly inhibited the proliferation and migration of OVCAR3 cells(P<0.05).Gyp-L effectively down-regulated the expression of 44 mirnas in OVCAR3 cells(P<0.05).After the intervention of Gyp-L,the level of miR-1291 was increased and the expression levels of ERRαand CPT1C were decreased in OVCAR3 cells(P<0.05).Gyp-L increased ROS level and decreased ATP content in OVCAR3 cells(P<0.05).The expressions of mitochondria-related genes(NRF1,PGC-1A,CYBA,and TFAM)were decreased(P<0.05),and glucose uptake ability was decreased(P<0.05).The changes of miR-1291/ERRα/CPT1C axis were found in clinical OC tissues(P<0.05).Conclusion The miR-1291/ERRα/CPT1C signaling pathway is involved in the occurrence and development of OC.Gyp-L inhibits the proliferation and metabolism of OVCAR3 cells by targeting ERRα/CPT1C signaling axis by highly expressing miR-1291,thereby controlling the occurrence and development of OC.
作者
崔鹏
朱敬轩
赵佼
王群
孙小扉
刘玉
杨潇
宋囡
CUI Peng;ZHU Jingxuan;ZHAO Jiao;WANG Qun;SUN Xiaofei;LIU Yu;YANG Xiao;SONG Nan(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Liaoning Cancer Hospital,Shenyang 110801,Liaoning,China;The Second Affiliated Hospital of Liaoning llniversity of Traditind Chinese Medicine,Shenyang 110034,Liaoning,China)
出处
《辽宁中医杂志》
北大核心
2025年第9期147-151,I0007-I0009,共8页
Liaoning Journal of Traditional Chinese Medicine
基金
国家自然科学基金面上项目(82274455)
辽宁省教育厅基本科研项目(JYTMS20231827,2024-JYTCB-008)。
