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prohibitin Protein Expression During Spermatogenesis in the Large Yellow Croaker, Larimichthys crocea

Journal of Ocean University of China 2022年第5期21卷 1251-1264页

作者： LIN Chenwen GAO Xinming NI Jie ZHANG Shengshuo LIU Cheng LUO Shengyu DU Chen ZHENG Xuebin HOU Congcong TANG Daojun ZHANG Chundan ZHU Junquan Key Laboratory of Applied Marine Biotechnology by the Ministry of Education School of Marine SciencesNingbo UniversityNingbo 315211China

Mitochondria are important for animals’fertility,and their morphologies and functions during spermatogenesis are un-der ***,the molecular mechanism that regulates the mitochondrial dynamic during spermatogenesis is still *** this study,the cytological features of spermatogenesis were investigated in Larimichthys *** addition,the structure and function of prohibitin(PHB),which is associated with mitochondrial structure and dynamic,was also *** full-length cDNA and protein(Lc-PHB)from the *** phb gene(Lc-phb)contained 1625 base pairs and 271 amino acids,***-PHB had a conserved primary structure that resulted in a transmembrane,SPFH(the analogous region of proteins stomat-ins,prohibitins,flotillins and HflK/C),and coiled-coil *** was detected at high levels in the muscle,liver,and heart,and at intermediate levels in the testis,gill,and ***-phb mRNA expression was detected in spermatogenic cells by fluorescence in situ *** immunofluorescence assay revealed that PHB protein was localized in the mitochondria during ***,PHB expression was detected in the perinuclear cytoplasm of spermatogonia,spermatocytes,and spermatids in the early developmental stage,and mainly localized on one side of the nuclei in the cytoplasm of spermatids in a middle developmental stage,and finally on the sperm *** blotting showed that PHB was located in the extracted mitochondria protein fraction but not in the cytoplasm protein fraction of ***,these results indicated that PHB was expressed in the mitochondria dur-ing *** addition,the study explained the mitochondrial dynamic during fish spermatogenesis and proposed a possi-ble relationship among PHB,spermatogenesis,and male fertility.

关键词： spermatogenesis ultrastructure prohibitin expression pattern subcellular localization

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prohibitin:a potential therapeutic target in tyrosine kinase signaling

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Signal Transduction and Targeted Therapy 2017年第1期2卷 10-17页

作者： Sudharsana Rao Ande Yang Xin Zi Xu Suresh Mishra Department of Internal Medicine University of ManitobaWinnipegManitobaCanada Department of Physiology&Pathophysiology University of ManitobaWinnipegManitobaCanada

prohibitin is a pleiotropic protein that has roles in fundamental cellular processes,such as cellular proliferation and mitochondrial housekeeping,and in cell-or tissue-specific functions,such as adipogenesis and immune cell *** different functions of prohibitin are mediated by its cell compartment-specific attributes,which include acting as an adaptor molecule in membrane signaling,a scaffolding protein in mitochondria,and a transcriptional co-regulator in the ***,the precise relationship between its distinct cellular localization and diverse functions remain largely *** evidence suggests that the phosphorylation of prohibitin plays a role in a number of cell signaling pathways and in intracellular ***,we discuss the known and potential importance of the site-specific phosphorylation of prohibitin in regulating these *** will discuss this in the context of new evidence from tissue-specific transgenic mouse models of prohibitin,including a mutant prohibitin lacking a crucial tyrosine phosphorylation *** conclude with the opinion that prohibitin can be used as a potential target for tyrosine kinase signal transduction-targeting therapy,including in insulin,growth factors,and immune signaling pathways.

关键词： prohibitin potential precise

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Up-regulation of prohibitin 1 is involved in the proliferation and migration of liver cancer cells

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Science China(Life Sciences) 2011年第2期54卷 121-127页

作者： XU ZhiBin WU JinHong ZHA XiLiang Institute of Biomedical Sciences Fudan University Shanghai 200032 China School of Agrieulture and Biology Bor Luh Food Safety Center Shanghai Jiaotong University Shanghai 200030 China Key Laboratory of Glycoconjugate Ministry of Health Department of Biochemistry and Molecular Biology Shanghai Medical College Fudan University Shanghai 200032 China

Although prohibitin 1(PHB) is known to be associated with tumors,there are few studies regarding the role of PHB in hepatocellular *** earlier glycoproteomics study of ours showed that PHB is over-expressed in MHCC97-H cells,a highly metastatic liver cancer cell line,and can be captured by wheat germ *** the present study,western blotting and reverse transcription-PCR experiments showed an approximately 2-fold up-regulation in the expression of PHB in MHCC97-H ***,PHB was not significantly up-regulated in MHCC97-L cells,a low-metastatic liver cancer cell *** PHB was over-expressed in MHCC97-L cells,cell proliferation was inhibited by 35% and migration increased about *** results of this study show that PHB is up-regulated in MHCC97-H cells and is associated with both proliferation and migration of liver cancer cells.

关键词： glycosylation lectin metastasis prohibitin up-regulation

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Research Progress in the Structure and Functions of prohibitin

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Journal of Reproduction and Contraception 2010年第2期21卷 117-124页

作者： Jia-xian OU Si-fei HUANG Hong CHEN Department of Anatomy & Histo-Embryology Shanghai Medical School Fudan University Shanghai 200032 China

prohibitin is named due to the negative regulatory role of its gene products in cell proliferation. prohibitin gene is located at q21 of chromosome 17 in human beings and the protein is found at mitochondria, nucleus and cytoplasm. Due to its size and ring-shaped structure, prohibitin protein defines functional subcompartments in mitochondria. Its subunits, PHB1 and PHB2, suppress cell proliferation as in the protein itself Nevertheless, recent investigation suggests that prohibitin protein enhances cell proliferation as well. It has also been found to suppress cell apoptosis by reducing cytochrome C release via the avoidance of mitochondrial crista remodeling which is facilitated through type 1 optic atrophy protein （OPAl）. Acting as a binding site for ubiquitin, prohibitin protein regulates protein degradation by proteasome. Examples are the degradations of sperm mitochondria in a fertilized ovum or those of an abnormal sperm.

关键词： prohibitin cell proliferation mitochondrial cristae type 1 optic atrophy protein(OPAl) cell apoptosis

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prohibitin蛋白与卵巢癌紫杉醇耐药相关性的初步研究

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中南大学学报（医学版） 2012年第12期37卷 1221-1227页

作者：唐娟曹兰琴易红汤参娥中南大学湘雅医院妇产科中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室长沙410008

目的:研究转染pshRNA/prohibitin(PHB)的RNA干扰对卵巢癌紫杉醇耐药细胞株的生物学特性的影响。方法:采用Western印迹及实时PCR方法验证卵巢癌紫杉醇耐药细胞株(SKOV3/Taxol-25)和敏感细胞株(SKOV3)中PHB蛋白与mRNA的表达差异。以脂质体... 详细信息

目的:研究转染pshRNA/prohibitin(PHB)的RNA干扰对卵巢癌紫杉醇耐药细胞株的生物学特性的影响。方法:采用Western印迹及实时PCR方法验证卵巢癌紫杉醇耐药细胞株(SKOV3/Taxol-25)和敏感细胞株(SKOV3)中PHB蛋白与mRNA的表达差异。以脂质体Lipofectamine2000为载体,将针对PHB基因设计的带有荧光蛋白的三靶特异性小发夹RNA干扰片段,即pshRNA/PHB427(PHB1实验组)、pshRNA/PHB248(PHB2实验组)、pshRNA/PHB136(PHB3实验组)瞬时转染耐药细胞株,另设阴性对照组。转染48 h后,用Western印迹及实时PCR方法检测PHB蛋白与mRNA的表达情况,筛选出沉默效果明显的两个片段(PHB1及PHB3)为实验组,与阴性对照组进行后续实验。用MTT及流式细胞术检测实验组及对照组的细胞增殖、紫杉醇IC50及凋亡情况。结果:耐药细胞株中PHB蛋白相对表达量及mRNA相对表达量(2–ΔΔCt)明显高于敏感细胞株(P<0.05)。PHB1和PHB3实验组的蛋白相对表达量及mRNA相对表达量(2–ΔΔCt)较阴性对照组明显降低(P<0.05)。转染48 h及72 h后PHB1及PHB3实验组细胞的增殖较阴性对照组明显减慢(P<0.05);干扰后72 h各实验组紫杉醇IC50较干扰前明显下降(P<0.05);转染48 h后PHB1及PHB3实验组细胞凋亡率较阴性对照组明显增加(P<0.05)。结论:针对PHB合成的shRNA能有效抑制耐药细胞株中PHB基因表达,沉默PHB基因后耐药细胞株的细胞增殖能力下降,凋亡率增加,对紫杉醇敏感性增加,提示PHB与卵巢癌紫杉醇耐药相关,干扰PHB基因的表达可能降低卵巢癌紫杉醇耐药性。

关键词： RNA干扰卵巢癌 prohibitin 紫杉醇耐药

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prohibitin在姜黄素诱导人成骨肉瘤MG-63细胞凋亡过程中的调控机制

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解剖学报 2015年第1期46卷 72-80页

作者：石松林赵振利王国红刘凡路锟杨玲韩荣李晓李祺福厦门大学医学院基础医学部福建厦门361102 河南农业大学林学院林学系郑州450002 新乡医学院基础医学系河南新乡453003

目的探讨姜黄素(curcumin)诱导成骨肉瘤细胞凋亡前后,prohibitin(PHB)蛋白的定位与表达变化,及PHB与凋亡相关基因产物的共定位关系。方法选择性抽提经姜黄素诱导处理前后的人成骨肉瘤MG-63细胞核基质蛋白,以蛋白质组学、免疫印迹法和激... 详细信息

目的探讨姜黄素(curcumin)诱导成骨肉瘤细胞凋亡前后,prohibitin(PHB)蛋白的定位与表达变化,及PHB与凋亡相关基因产物的共定位关系。方法选择性抽提经姜黄素诱导处理前后的人成骨肉瘤MG-63细胞核基质蛋白,以蛋白质组学、免疫印迹法和激光扫描共焦显微技术对PHB在核基质中存在、分布及其与凋亡相关基因产物在MG-63细胞中的共定位关系进行了研究。结果双向凝胶电泳、质谱鉴定和免疫印迹法结果显示,PHB存在于MG-63细胞核基质蛋白组分中,细胞凋亡后在细胞核基质蛋白中表达下调。免疫荧光显微镜观察显示,PHB定位于MG-63细胞核基质纤维上,经姜黄素处理后出现分布位置与表达水平的变化。激光扫描共焦显微镜观察结果显示,PHB在MG-63细胞凋亡过程中与Bax、Bcl-2、Fas、P53、c-Myc和Rb等基因产物具有共定位关系,且共定位区域发生了变化。结论 PHB是一种核基质结合蛋白;PHB在MG-63细胞凋亡过程中的表达与分布变化,及其与细胞凋亡相关基因的共定位关系,对MG-63细胞凋亡具有重要影响。

关键词： prohibitin 核基质人成骨肉瘤MG-63细胞姜黄素免疫印迹法

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prohibitin特异性MiRRNAi表达载体构建及其干扰效果测定

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中国应用生理学杂志 2009年第1期25卷 139-144页

作者：郭东升王新兴刘晓华战锐袁聚祥钱令嘉华北煤炭医学院预防医学系河北唐山063000 军事医学科学院卫生学环境医学研究所应激医学实验室天津300050

目的:构建携带prohibitin(PHB-1)基因的MiR RNAi真核表达载体pcDNA^(TM)6.2-GW/EmGFP-MiR,并观察其转染HEK293细胞株前后PHB-1的表达变化。方法:根据GenBank中prohibitin的序列,设计特异的两条互补的单链寡核苷酸退火后形成双链,克隆至pcDNA^(TM)6.2-GW/EmGFP-MiR质粒缺口末端,连接在质粒上生成含MiR RNAi pcDNA^(TM) 6.2-GW/EmGFP-MiR-PHB载体,测序鉴定后,用脂质体将重组子转染至HEK293细胞中,用Western blotting检测干扰后HEK293细胞内PHB-1表达的变化。结果:将目的序列成功连接到载体上,并经测序分析证实载体构建成功。Western blotting检测结果证实构建的PHB-1 MiR RNA表达重组体可显著抑制HEK293细胞内PHB-1的表达。结论:成功构建了携带PHB-1基因的MiR RNAi真核表达载体。

关键词： prohibitin HEK293细胞 RNA干扰

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prohibitin在低胆固醇培养人前列腺癌细胞系PC-3中表达上调及其可能机制

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中国病理生理杂志 2011年第11期27卷 2116-2119页

作者：董培李永红尧凯刘卓炜周芳坚华南肿瘤学国家重点实验室中山大学肿瘤防治中心泌尿外科广东广州510060

目的:探讨prohibitin(PHB)表达上调在低胆固醇培养抑制前列腺癌PC-3细胞增殖中的作用及对前列腺癌进展的影响。方法:PC-3细胞分别培养于普通和低胆固醇培养基中48 h后,real-time PCR鉴定PHB mRNA的差异表达。克隆PHB基因启动子片段,构... 详细信息

目的:探讨prohibitin(PHB)表达上调在低胆固醇培养抑制前列腺癌PC-3细胞增殖中的作用及对前列腺癌进展的影响。方法:PC-3细胞分别培养于普通和低胆固醇培养基中48 h后,real-time PCR鉴定PHB mRNA的差异表达。克隆PHB基因启动子片段,构建萤光质粒并转染至PC-3细胞,检测细胞培养基中胆固醇变化对于PHB基因启动子萤光素酶表达影响。结果:低胆固醇培养基组中PHB表达上调约19倍,在低胆固醇培养基的前列腺癌PC-3细胞中PHB基因启动子质粒萤光素酶表达显著增强,同普通培养对照组相比有显著差异。结论:PHB在低胆固醇培养前列腺癌PC-3细胞中表达显著增强,这可能是前列腺癌细胞在不利于增殖条件下的一种自我保护调节机制。

关键词：前列腺肿瘤胆固醇 prohibitin 启动子

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Positional and expressive alteration of prohibitin during the induced differentiation of human hepatocarcinoma SMMC-7721 cells

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World Journal of Gastroenterology 2008年第32期14卷 5008-5014页

作者： Dong-Hui Xu Jian Tang Qi-Fu Li Song-Lin Shi Xiang-Feng Chen Ying Liang Department of Hepatic Biliary Pancreatic Vascular Surgery 1st Hospital of Xiamen Fujian Medical University Xiamen 361003 Fujian Province China The Key Laboratory of Chinese Ministry of Education for Cell Biology and Tumor Cell Engineering School of Life Sciences Xiamen University Xiamen 361005 Fujian Province China

AIM： To explore the existence and distribution of prohibitin （PHB） in nuclear matrix and its co-localization with products of some related genes during the differentiation of human hepatocarcinoma SNMC-7721 cells. METHODS： The nuclear matrix of the SMMC-7721 cells cultured with or without 5 × 10^-3 mmol/L hexamethylene bisacetamide （HNBA） was selectively extracted. Western blot was used to analyze the expression of PHB in nuclear matrix; immunofluorescence microscope observation was used to analyze the distribution of PHB in cell. LCSM was used to observe the co-localization of PHB with products of oncogenes and tumor suppressor genes. RESULTS： Western blot analysis showed that PHB existed in the composition of nuclear matrix proteins and was down-regulated by HMBA treatment. Immunofluorescence observation revealed that PHB existed in the nuclear matrix, and its distribution regions and expression levels were altered after HMBA treatment. Laser scanning confocal microscopy revealed the co-localization between PHB and the products of oncogenes or tumor repression genes including c-los, c-myc, p53 and Rb and its alteration of distributive area in the cells treated by HMBA. CONCLUSION： These data confirm that PHB is a nuclear matrix protein, which is located in the nuclear matrix, and the distribution and expression of PHB and its relation with associated genes may play significant roles during the differentiation of SMMC-7721 cells.

关键词： prohibitin Nuclear matrix SMMC-7721 cells Hexamethylamine Bisacetamide Cell differentiation

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Dingxin Recipe(定心方) Prevents Ischemia/Reperfusion-Induced Arrhythmias via Up-regulating prohibitin and Suppressing Inflammatory Responses

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Chinese Journal of Integrative Medicine 2012年第2期18卷 120-129页

作者：贾钰华张云仙李丽君刘亚伟黎春华符秀琼曾平吴伟康孙学刚 The Key Laboratory of Molecular Biology State Administration of Traditional Chinese MedicineSchool of Traditional Chinese MedicineSouthern Medical University Fuda Cancer Hospital Guangdong General Hospital Guangdong Academy of Medical Sciences The Institute of Integrated Traditional Chinese and Western Medicine Sun Yatsen University

Objective： To identify the underlying mechanisms of the protective effects of Dingxin Recipe （定心方, DXR）, a Chinese compound prescription that has been used clinically in China for more than 20 years, on ischemia/reperfusion （I/R）-induced arrhythmias in rat model. Methods： A total of 30 rats were randomly divided into three groups： sham group, I/R group, and DXR-pretreated I/R （DXR-I/R） group. Rats in the DXR- I/R group were intragastrically administrated with DXR （12.5 g/kg per day） for consecutive 7 days, while rats in the sham and I/R groups were administrated with normal saline. Arrhythmias were introduced by I/R and electrocardiograms （ECG） were recorded. Two-dimensional （2-D） polyacrylamide gel electrophoresis and matrix- assisted laser desorption ionization time-of-flight mass spectrometry （MALDI-TOF MS） were used to identify differentially expressed proteins. Immunohistochemistry, real-time quantitative polymerase chain reaction （RQ- PCR）, Western blot, and enzyme-linked immunosorbent assay （ELISA） were performed to analyze proteins obtained in the above experiments. Results： DXR significantly reduced the incidence and mean duration of ventricular tachycardia and ventricular fibrillation and dramatically decreased the mortality, as well as arrhythmia score, compared with those of the I/R group. Among successfully identified proteins, prohibitin （PHB） and heart fatty acid binding protein （hFABP） were up-regulated in DXR-pretreated I/R rats compared with those of the I/R rats. In addition, compared with the I/R group, the level of glutathione （GSH） was elevated accompanied by reduced expressions of interleukin-6 （IL-6） and neutrophil infiltration in I/R rats with DXR pretreatment. Conclusions： DXR could alleviate I/R-induced arrhythmias, which might be related to increased expression of PHB. The enhanced expression of PHB prevented against the depletion of GSH and consequently inhibited apoptosis of cardiomyocytes. Furthermore, up-regulation of PHB might amelio

关键词： Dingxin Recipe proteomics arrhythmias prohibitin ischemiaJreperfusion inflammation

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