OPA1在乳腺癌组织的表达特征及在ER阳性乳腺癌细胞中的生物学功能研究  

Expression characterization OPA1 expression in breast cancer tissues and its biological function in ER-positive breast cancer cells

作　　者：黄程鑫 陈莉 刘伊楚 王水良 赖晓凤 Chengxin Huang;Li Chen;Yichu Liu;Shuiliang Wang;Xiaofeng Lai(Department of Laboratory Medicine,Fujian University of Traditional Chinese Medicine,Fuzong Teaching Hospital(900th Hospital),Fuzhou 350025,China;Fujian Key Laboratory of Aptamer Technology,Fujian Clinical Research Center for Aptamer-based Precision Diagnostics and Clinical Medicine,Fuzhou 350025,China;Department of Laboratory Medicine,Fujian Medical University,Fuzong Clinical Medical College(900th Hospital),Fuzhou 350025,China;Department of Laboratory Medicine,Dongfang Hospital Affiliated to Xiamen University(900th Hospital),Fuzhou 350025,China)

机构地区：[1]福建中医药大学福总教学医院(第九〇〇医院)检验科,福州350025 [2]福建省适配体技术重点实验室,福建省适配体精准检验临床医学研究中心,福州350025 [3]福建医科大学福总临床医学院(第九〇〇医院)检验科,福州350025 [4]厦门大学附属东方医院(第九〇〇医院)检验科,福州350025

出　　处：《中华细胞与干细胞杂志（电子版）》2024年第5期275-284,共10页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：国家自然科学基金(81902712);福建省自然科学基金(2021J011269、2023J011345);中国人民解放军联勤保障部队第900医院基金(2023SA01、2023SA03、2023QN09)。

摘　　要：目的分析视神经萎缩蛋白1(OPA1)在乳腺癌的表达情况及其对雌激素受体(ER)阳性乳腺癌细胞的生物学功能影响,为乳腺癌治疗寻求潜在新靶点。方法分别用公开数据信息(HPA和GSE115144)以及ER阳性乳腺癌患者组织芯片分析OPA1在乳腺癌和正常/癌旁组织中的表达差异,通过GEPIA和TCGA数据库分析OPA1表达与乳腺癌患者生存期的相关性。在细胞实验中,分别使用OPA1质粒和小干扰RNA(siRNA)对MCF7和T47D细胞进行转染,通过RT-qPCR和Western blot检测OPA1的mRNA和蛋白相对表达水平。采用CCK-8实验与平板克隆形成实验分别检测细胞增殖及克隆形成能力,流式细胞术检测活性氧(ROS)水平、线粒体膜电位(JC-1法)和细胞凋亡。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析,组间两两比较采用Dunnett-t检验。结果与癌旁组织相比,OPA1在乳腺癌组织中高表达(P<0.01),并且OPA1高表达与乳腺癌患者临床预后不良相关(P<0.05)。细胞实验中,过表达OPA1促进细胞增殖和克隆形成,降低ROS水平[(37.37±1.48)%比(62.51±2.66)%,P<0.001],同时线粒体膜电位增加[JC-1单体比例降低(7.32±1.84)%比(17.6±3.35)%,P<0.01];而抑制OPA1能够抑制细胞增殖和克隆形成,增加ROS水平[(77.81±2.37)%比(58.37±1.36)%,P<0.001]与降低线粒体膜电位[JC-1单体比例升高(28.13±3.47)%比(15.96±1.14)%,P<0.01],并且促进细胞凋亡[(20.10±1.20)%比(3.85±0.76)%,P<0.001]。结论OPA1在乳腺癌中高表达且与临床预后不良相关,抑制OPA1表达可抑制ER阳性乳腺癌细胞增殖和克隆形成能力,同时降低线粒体膜电位,影响线粒体功能,诱导ROS生成最终促进细胞凋亡。Objective To analyze the expression of Optic Atrophy 1(OPA1)in breast cancer and its effect on the biological function of estrogen receptor(ER)-positive breast cancer cells,seeking potential new targets for breast cancer therapy.Methods Expression differences of OPA1 in breast cancer and normal/paracancerous tissues were analyzed using publicly available database information(HPA and GSE115144)and tissue microarrays from ER-positive breast cancer patients,respectively.The correlation between OPA1 expression and survival in breast cancer patients was analyzed by the GEPIA and TCGA databases.In cellexperiments,MCF7 and T47D cells were transfected with OPA1 plasmid and small interfering RNA(siRNA),respectively,and the relative expression levels of mRNA and protein of OPA1 were detected by RT-qPCR and Western blot.Cell proliferation and clone formation ability were detected by CCK-8 assay and plate clone formation assay,respectively.Reactive oxygen species(ROS)level,mitochondrial membrane potential(JC-1 method)and apoptosis rate were detected by flow cytometry.Two independent samples t-test was used for comparison between groups.Differences among groups were compared by one-way ANOVE analysis,and Dunnett-t test was used for pair-to-group comparisons.Results OPA1 was highly expressed in breast cancer tissues compared with paracancerous tissues(P<0.01),and high OPA1 expression was associated with poor clinical prognosis in breast cancer patients(P<0.05).In cell experiments,overexpression of OPA1 promoted cell proliferation and clone formation and decreased the ROS levels[(37.37±1.48)%vs(62.51±2.66)%,P<0.001],along with an increase in mitochondrial membrane potential[decrease in the proportion of JC-1 monomers,(7.32±1.84)%vs(17.6±3.35)%,P<0.01];while inhibition of OPA1 was able to inhibit cell proliferation and clone formation,increase ROS levels[(77.81±2.37)%vs(58.37±1.36)%,P<0.001],decreasing mitochondrial membrane potential[elevated proportion of JC-1 monomer,(28.13±3.47)%vs(15.96±1.14)%,P<0.01],and promote ap

关 键 词：乳腺癌 视神经萎缩蛋白1 细胞增殖 细胞凋亡 线粒体 

分 类 号：R737.9[医药卫生—肿瘤]