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mkk7-mediated phosphorylation of JNKs regulates the proliferation and apoptosis of human spermatogonial stem cells

World Journal of Stem Cells 2021年第11期13卷 1797-1812页

作者： Zeng-Hui Huang Chuan Huang Xi-Ren Ji Wen-Jun Zhou Xue-Feng Luo Qian Liu Yu-Lin Tang Fei Gong Wen-Bing Zhu Institute of Reproductive and Stem Cell Engineering Central South UniversityChangsha 410008Hunan ProvinceChina Department of Reproductive Center Reproductive and Genetic Hospital of CITIC-XiangyaChangsha 410008Hunan ProvinceChina Department of Sperm Bank Reproductive and Genetic Hospital of CITICXiangyaChangsha 410008Hunan ProvinceChina

BACKGROUND Human spermatogonial stem cells(SSCs)are the basis of ***,little is known about the developmental regulatory mechanisms of SSC due to sample origin and species *** To investigates the mechanisms involved in the proliferation of human *** The expression of mitogen-activated protein kinase kinase 7(mkk7)in human testis was identified using immunohistochemistry and western blotting(WB).mkk7 was knocked down using small interfering RNA,and cell proliferation and apoptosis were detected by WB,EdU,cell counting kit-8 and fluorescenceactivated cell *** bioinformatic analysis,the interaction of mkk7 with c-Jun N-terminal kinases(JNKs)was verified by protein co-immunoprecipitation and *** phosphorylation of JNKs was inhibited by SP600125,and the phenotypic changes were detected by WB,cell counting kit-8 and fluorescenceactivated cell *** mkk7 is mainly expressed in human SSCs,and mkk7 knockdown inhibits SSC proliferation and promotes their ***7 mediated the phosphorylation of JNKs,and after inhibiting the phosphorylation of JNKs,the phenotypic changes of the cells were similar to those after mkk7 *** expression of mkk7 was significantly downregulated in patients with abnormal spermatogenesis,suggesting that abnormal mkk7 may be associated with spermatogenesis *** mkk7 regulates the proliferation and apoptosis of human SSC by mediating the phosphorylation of JNKs.

关键词： mkk7 Spermatogonial stem cell Proliferation Apoptosis JNKs

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mkk7-Mediated Signaling Pathways in Arabidopsis

MKK7-Mediated Signaling Pathways in Arabidopsis

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中国遗传学会植物遗传和基因组学专业委员会2009年学术研讨会

作者： Baohua Li,Weiyan Jia,Huanzhong Wang,Yonghong Wang,and Jiayang Li~* State Key Laboratory of Plant Genomics and National Center for Plant Gene Research,Institute of Genetics and Developmental Biology,Chinese Academy of Sciences,Beijing 100101,China

Mitogen-activated protein kinase(MAPK) cascades are universal signal transduction modules in *** plants,MAPK cascades are involved in diverse biological processes,including biotic and abiotic stresses,hormonal actions... 详细信息

关键词： Arabidopsis mkk7 polar auxin transport signaling

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mkk7/JNK途径促进HCT116细胞恶性生长

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军事医学 2013年第9期37卷 666-670页

作者：朱婷张大林王晶黎燕沈倍奋张纪岩王庆阳中南大学基础医学院免疫学系长沙410078 军事医学科学院基础医学研究所北京100850

目的观察mkk7/JNK途径对HCT116细胞恶性生长的影响。方法建立稳定敲低mkk7与JNK的细胞株,Western印迹实验检测JNK与mkk7的表达,集落形成技术检测HCT116细胞体外增殖。通过裸鼠荷瘤实验分析稳定敲低mkk7与JNK细胞株体内恶性生长能力。结... 详细信息

目的观察mkk7/JNK途径对HCT116细胞恶性生长的影响。方法建立稳定敲低mkk7与JNK的细胞株,Western印迹实验检测JNK与mkk7的表达,集落形成技术检测HCT116细胞体外增殖。通过裸鼠荷瘤实验分析稳定敲低mkk7与JNK细胞株体内恶性生长能力。结果稳定敲低mkk7与JNK细胞株集落形成能力显著降低,成瘤能力明显减弱。结论 mkk7/JNK途径促进了HCT116细胞恶性生长。

关键词： mkk7 JNK HCT116细胞结肠肿瘤恶性生长

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RACK1 directly interacts with mkk7 and mediates JNK hyper-activation in human hepatocellular carcinoma

RACK1 directly interacts with MKK7 and mediates JNK hyper-ac...

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中国的遗传学研究——遗传学进步推动中国西部经济与社会发展——2011年中国遗传学会大会

作者： Yuanyuan Guo,~(1,2) Jing Wang,~1 Qingyang Wang,~1 Jiannan Feng,~1 Ming Lv,~1 Xueying Zhang,~1 Yuanfang Ma,~(1,2) Xinying Li,~1 Yan Li,~1 Beifen Shen,~1 and Jiyan Zhang~1 1 Department of Molecular Immunology,Institute of Basic Medical Sciences,27 Taiping Road,Beijing 100850,P R China.2 Laboratory of Cellular and Molecular Immunology,Henan University,Kaifeng 475001,P R China.

Extensive studies revealed an essential protumorigenic role of constitutively activated JNK(c-Jun N-terminal protein kinase) in hepatocellular carcinoma(HCC).RACK1(the receptor of activated C kinase 1),an adaptor protein implicated in the regulation of multiple signaling pathways,was also shown upregulated in *** is of importance to investigate the possible reciprocal regulation between the JNK pathway and RACK1 in ***,we report that the protein level of RACK1 showed correlation with JNK activation in human ***1 overexpression led to enhanced mkk7 activation as well as JNK-c-Jun activation in various cell types including human hepatoma cells,whereas RACK1 knock down exhibited opposite *** two-hybrid screening identified RACK1 as a novel mkk7-interacting ***1 mutants defective of anchoring mkk7 showed reduced ability to enhance JNK activation and human hepatoma cell growth in vitro and in nude mice as compared with wild type ***-αinduced RACK1 upregulation,which was blocked by JNK ***,our data suggest an inflammation-related feed forward mechanism in HCC:Exposure to TNF-αleads to augmented RACK1 expression via the activation of the JNK *** RACK1 expression in turn contributes to JNK hyper-activation through,at least partially,directly interacting with mkk7.

关键词： RACK1 interaction mkk7 JNK hepatocellular carcinoma

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Neddylation controls basal mkk7 kinase activity in breast cancer cells

Neddylation controls basal MKK7 kinase activity in breast ca...

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第十届全国免疫学学术大会

作者： Jing Wang Yujun Pei Ting Zhu Qingyang Wang Ye Wu Guihua Qiu Dalin Zhang Beifen Shen Jiyan Zhang Beijing Institute of Basic Medical Sciences

The JNK pathway has been implicated in mammary tumor ***,the molecular mechanisms regulating JNK activity in breast cancer cells remain ***,we report that mkk7,a JNK-specific MAP2K,is covalently conjugated with the ubiquitin-like modifier NEDD8 in human breast cancer *** of NEDD8 or UBA3,a component of the E1 heterodimer,results in enhanced basal JNK *** augmented basal JNK phosphorylation via ectopic mkk7 expression leads to reduced anchorage-independent growth and more apoptosis in response to TRAIL,the neddylation system might contribute to mammary tumor development partially through limiting basal JNK *** exploration suggests that the neddylation system inhibits the basal kinase activity of ***7 co-precipitated with a fragment of Ran-binding protein 2(Ran BP2),a large multimodular and pleiotropicprotein which has been recognized as a SUMO E3 *** of Ran BP2 attenuates mkk7 neddylation and augments basal JNK phosphorylation without affecting the neddylation of Cullins,whereas ectopic expression of a Ran BP2 fragment possessing SUMO E3 activity(Ran BP2DFG)manifests the opposite *** vitro neddylation assays confirm that Ran BP2DFG works as the neddylation E3 ligase for *** basal kinase activity of endogenous mkk7 increases upon Ran BP2 knockdown but decreases upon the ectopic expression of Ran ***,purified mkk7 shows reduced basal kinase activity after in vitro *** together,our data suggest that mkk7undergoes neddylation in human breast cancer cells,which limits its basal kinase activity.

关键词： mkk7 kinase activity breast cancer neddylation oncogenic growth

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Funtional analysis of mkk7-MPKs in plant development

Funtional analysis of MKK7-MPKs in plant development

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从植物科学到农业发展——2012全国植物生物学大会

作者： Weiyan Jia,Baohua Li,Shujia Li,Yonghong Wang and Jiayang Li~* State Key Laboratory of Plant Genomics and National Center for Plant Gene Research,Institute of Genetics and Developmental Biology,Chinese Academy of Sciences,Beijing,China.

Mitogen-activated protein kinase(MAPK) cascades play an important role in transducing environmental and developmental signals into adaptive and programmed *** higher plant,MAPK cascades are involved in a broad spectrum of signals,which include biotic and abiotic stresses,and hormone-mediated *** previous study have characterized a semidominant Arabidopsis thaliana bushy and dwarf1(bud1)mutant. Molecular genetic analysis indicated that the budl phenotype is a result of increased expression of Arabidopsis MAPKINASE KINASE7(mkk7),a member of plant mitogen-activated protein kinase kinase group *** showed that BUD1/mkk7 is a functional kinase that affects plant architecture and negatively regulates polar auxin transport. The Arabidopsis genome encodes a large number of MAPK signaling components,with more than 60 members of MAPKKK,10 members of mkk and 20 members of MAPK. The mismatch between the numbers of mkks and their putative MPK substrates suggests that individual mkks should have the capacity to active more than one *** elucidate the specificity of BUD1/mkk7 and its downstream MPKs in plant development,we first systemically screened the downstream substrates of mkk7 in ***,we characterized the function of BUD1/mkk7-MPKs in *** study will provide evidence to explain how BUD1/mkk7 acts multiple roles in plant development.

关键词： mkk7 MPK bud1 plant architecture Arabidopsis thaliana

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Mitogen-activated protein kinase cascade mkk7-MPK6 plays important roles in plant development and regulates shoot branching by phosphorylating PIN1 in Arabidopsis

Mitogen-activated protein kinase cascade MKK7-MPK6 plays imp...

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2016年全国植物生物学大会

作者： Weiyan Jia Baohua Li Shujia Li Yan Liang Xiaowei Wu Mei Ma Jiyao Wang Yueyue Cai Yuanya Zhang Yingchun Wang Jiayang Li Yonghong Wang State Key Laboratory of Plant Genomics and National Center for Plant Gene Research (Beijing) CAS Center for Excellence in Molecular Plant SciencesInstitute of Genetics and Developmental BiologyChinese Academy of Sciences Department of Human Oncology University of WisconsinSchool of Medicine and Public Health Department of Plant Sciences University of California State Key Laboratory of Molecular Developmental Biology Institute of Genetics and Developmental BiologyChinese Academy of Sciences

Emerging evidences exhibit that mitogen-activated protein kinase(MAPK) signaling pathways are connected with many aspects of plant development. The complexity of MAPK cascades raises challenges not only to identify the MAPK module in planta but also to define the specific role of an individual module. Our previous study have characterized an Arabidopsis bushy and dwarf1(bud1) mutant,in which the mkk7 was constitutively activated,resulting in multiple phenotypic alterations and activated the mobile signal of systemic acquired resistance. To elucidate the specificity of mkk7 and its downstream MAPKs in multiple biological processes,we systemically screened the downstream substrates of mkk7 in vitro. We found that MPK3 and MPK6 are the substrates for phosphorylation by mkk7 in planta. Genetic analysis showed that mkk7-MPK6 cascade is specifically responsible for the regulation of shoot branching,hypocotyl gravitropism,filament elongation,and lateral root formation. We further demonstrated that the mkk7-MPK6 cascade controls shoot branching by phosphorylating Ser 337 on PIN1,which affects the basal localization of PIN1 in xylem parenchyma cells and polar auxin transport in the primary stem. Our findings specify the functions of the mkk7-MPK6 cascade,and explain how mkk7-MPK6 signaling pathway regulates polar auxin transport(PAT) through the specific substrate PIN1 to determine shoot branching in Arabidopsis,establishing a molecular link between the MAPK cascade and auxin-regulated plant development.

关键词： mkk7 MPK6 phosphorylation PIN1 shoot branching

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RNA干扰沉默mkk7对SH-SY5Y细胞凋亡的影响

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南京医科大学学报（自然科学版） 2015年第5期35卷 656-661页

作者：赖露颖张庆国李乐赵伟雷洪伊姜珊张婧徐世元南方医科大学珠江医院麻醉科广东广州510282

目的 :探讨RNA干扰沉默丝裂原激活的蛋白激酶激酶(mitogen-activated protein kinase kinase,mkk7)基因对SHSY5Y细胞凋亡的影响。方法:设计并合成针对mkk7的小干扰RNA(small interference RNA,si RNA)3条(mkk7si RNA-1、mkk7si RNA-2、mkk7si RNA-3)及与mkk7基因无同源性的带有红色荧光的阴性对照si RNA(negative control si RNA),在lipofectamine 2000介导下转染SH-SY5Y细胞,荧光显微镜下观察转染效果,RT-PCR观察mkk7 m RNA表达及Western blot检测mkk7蛋白的表达,进而筛选出转染效果好的mkk7si RNA,CCK-8检测各组细胞活力,Hochest 33258检测各组细胞凋亡染色,流式细胞仪检测细胞凋亡率,Western blot检测p-JNK、凋亡蛋白cleaved caspase-3的变化。结果:1 mkk7si RNA-3组的mkk7 m RNA表达最低,mkk7蛋白表达最低;2与空白对照组(control group)比较,mkk7si RNA-3组的细胞活力无统计学差异,pJNK、cleaved caspase-3水平降低,凋亡率降低。结论:RNA干扰沉默mkk7可通过下调JNK磷酸化进而抑制SH-SY5Y细胞的凋亡。

关键词： SH-SY5Y细胞 RNA干扰 mkk7 凋亡

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mkk7在类风湿关节炎滑膜中的表达

MKK7在类风湿关节炎滑膜中的表达

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作者：刘一秀中国医科大学

学位级别：硕士

目的类风湿性关节炎是一种慢性进行性的自身免疫性疾病,严重威胁人类的健康。现在人们认为细胞因子网络调节异常是它的免疫病理机制和效应。以前的研究已证实通过对这个网络上的分子进行抑制来治疗本病已显示出特异性好,效果显著的特点... 详细信息

目的类风湿性关节炎是一种慢性进行性的自身免疫性疾病,严重威胁人类的健康。现在人们认为细胞因子网络调节异常是它的免疫病理机制和效应。以前的研究已证实通过对这个网络上的分子进行抑制来治疗本病已显示出特异性好,效果显著的特点。JNK这条通路在类风湿性关节炎中占有重要地位,且其中游的分子又仅有mkk4/7两个,故可以认为对这个位点的抑制是简单,且效果名显的。本实验通过检测丝裂酶原活性蛋白激酶激酶7（mkk7）在类风湿患者,骨性关节炎患者,正常人滑膜中的表达情况。探讨mkk7与类风湿关节炎发病的相关性。为进一步明确类风湿关节的发病机制奠定基础,为临床治疗提供一定的理论依据。方法实验组选取中国医科大学第一附属医院骨科2006年7月～2007年9月行膝关节腔清理术和关节置换术的RA患者10例,取其膝关节滑膜作为标本。。RA:其中男4例,女6例,年龄38岁～68岁,平均52.5岁。病程4个月～14年,平均45.2个月。对照组选取中国医科大学第一附属医院骨科2006年7月～2007年9月行膝关节腔清理术,探查术和关节置换术的OA患者7例,正常人6例,分别取滑膜组织作为标本。OA:男3例,女4例,年龄42～65,平均53岁。病程2个月～20年,平均70月。正常组:男3例,女3例,年龄28～43,平55岁。病程1个月～6个月,平均4个月。以计算机为辅助合成两条引物,以RT-PCR技术检测三组滑膜中的mkk7 mRNA的含量并进行比较,Western-blot技术检测mkk7在RA组和两对照的表达及比较他们之间的差异（以磷酸化和非磷酸化的mkk7两种抗体进行检测,以便说明这种蛋白的活性）。用计算机软件计算两种结果的灰度值,进行对比（图像分析系统LeicaQ550 IW:灰度分级0～255级,0级为最深,255级为最浅）。所有数据采用SPSS11.5 ForWindows统计软件处理,阳性率比较采用X检验,以P＜0.05作为有统计学意义的检验水准。结果RT-PCR结果显示:mkk7 mRNA的含量在实验组和对照组中均有表达,且实验组与对照组之间的差别具有统计学意义（P＜0.05）。Western-Blot结果显示:以非磷酸化的mkk7抗体进行蛋白定量的三组标本中均有mkk7的表达,实验组与对照组之间的差异有统计学意义（P＜0.05）。以磷酸化的mkk7抗体进行蛋白定量,见实验组仍有大量蛋白表达,但两对照组蛋白含量很少。统计学上实验组与对照组之间的有明显差异（P＜0.05）,对比两种抗体灰度值,证明mkk7在类风湿关节炎中的活性明显高于两对照组。（灰度值分析见三格表）结论本研究表明mkk7在RA的滑膜中含量多且活性高,这可能与RA的发病密切相关,通过对其阻断可能会部分缓解本病的病情。

关键词：类风湿关节炎 mkk7 信号转导途径

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RAGE-mkk4/mkk7-JNK1通路在2型糖尿病合并肝细胞癌中的作用

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广东医学 2017年第10期38卷 1509-1514页

作者：蒙丽恒黄耀李世春秦映芬广西医科大学第一附属医院内分泌科广西南宁530021

目的探讨晚期糖基化终末产物受体(RAGE)、丝裂原活化蛋白激酶激酶4(mkk4)、丝裂原活化蛋白激酶激酶7(mkk7)及c-Jun氨基末端蛋白激酶1(JNK1)在2型糖尿病合并肝细胞癌中的表达及意义。方法收集肝胆外科手术切除的2型糖尿病合并肝细胞癌(D... 详细信息

目的探讨晚期糖基化终末产物受体(RAGE)、丝裂原活化蛋白激酶激酶4(mkk4)、丝裂原活化蛋白激酶激酶7(mkk7)及c-Jun氨基末端蛋白激酶1(JNK1)在2型糖尿病合并肝细胞癌中的表达及意义。方法收集肝胆外科手术切除的2型糖尿病合并肝细胞癌(DMHC组)癌组织和相应癌旁组织标本18例及30例肝细胞癌(HCC组)癌组织和相应癌旁组织,用Western blot法检测其RAGE、mkk4、mkk7及JNK1的蛋白表达情况。结果 (1)DMHC及HCC组癌组织中RAGE、mkk7及JNK1蛋白表达均较相应癌旁组织高,mkk4蛋白表达水平均较相应癌旁组织表达低,差异均有统计学意义(Pmkk7和JNK1蛋白表达高于HCC组癌组织,差异有统计学意义(Pmkk7和JNK1蛋白表达水平均较高分化的癌组织高,而DMHC组低分化的癌组织mkk7和JNK1蛋白表达水平较高分化的癌组织表达高,差异均有统计学意义(Pmkk4蛋白表达水平较未发生转移者降低,差异有统计学意义(Pmkk7、RAGE与JNK1、mkk7与JNK1蛋白表达水平均呈显著正相关关系(Pmkk4降低可能是单纯肝细胞癌及2型糖尿病合并肝细胞癌的危险因素之一;糖尿病状态下mkk7、JNK1蛋白更高的表达可能促进了2型糖尿病时肝细胞癌的发生和发展,mkk4降低可能导致了肝癌细胞的转移。mkk7和JNK1可能在一定水平上受RAGE的调控。

关键词： RAGE mkk4 mkk7 JNK1 糖尿病肝细胞癌

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