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Cucurbitacin B-induced G2/M cell cycle arrest of conjunctival melanoma cells mediated by grp78-FOXM1-KIF20A pathway

Acta Pharmaceutica Sinica B 2022年第10期12卷 3861-3876页

作者： Jinlian Wei Xin Chen Yongyun Li Ruoxi Li Keting Bao Liang Liao Yuqing Xie Tiannuo Yang Jin Zhu Fei Mao Shuaishuai Ni Renbing Jia Xiaofang Xu Jian Li State Key Laboratory of Bioreactor Engineering Shanghai Frontiers Science Center of Optogenetic Techniques for Cell MetabolismFrontiers Science Center for Materiobiology and Dynamic ChemistryShanghai Key Laboratory of New Drug DesignSchool of PharmacyEast China University of Science and TechnologyShanghai 200237China Department of Ophthalmology Ninth People’s HospitalShanghai Jiao Tong University School of MedicineShanghai 200025China Cancer Institute Longhua HospitalShanghai University of Traditional Chinese MedicineShanghai 200032China Yunnan Key Laboratory of Screening and Research on Anti-pathogenic Plant Resources from West Yunnan College of PharmacyDali UniversityDali 671000China Clinical Medicine Scientific and Technical Innovation Center Shanghai Tenth People’s HospitalTongji University School of MedicineShanghai 200092China Key Laboratory of Tropical Biological Resources of Ministry of Education College of PharmacyHainan UniversityHaikou 570228China

Conjunctival melanoma(CM) is a rare and fatal malignant eye tumor. In this study, we deciphered a novel anti-CM mechanism of a natural tetracyclic compound named as cucurbitacin B(CuB). We found that CuB remarkably inhibited the proliferation of CM cells including CM-AS16,CRMM1, CRMM2 and CM2005.1, without toxicity to normal cells. CuB can also induce CM cells G2/M cell cycle arrest. RNA-seq screening identified KIF20A, a key downstream effector of FOXM1 pathway, was abolished by CuB treatment. Further target identification by activity-based protein profiling chemoproteomic approach revealed that grp78 is a potential target of CuB. Several lines of evidence demonstrated that CuB interacted with grp78 and bound with a Kdvalue of0.11 μmol/L. Furthermore, ATPase activity evaluation showed that CuB suppressed grp78 both in human recombinant grp78 protein and cellular lysates. Knockdown of the grp78 gene significantly induced the downregulation of FOXM1 and related pathway proteins including KIF20A, underlying an interesting therapeutic perspective. Finally, CuB significantly inhibited tumor progression in NCG mice without causing obvious side effects in vivo. Taken together, our current work proved that grp78-FOXM1-KIF20A as a promising pathway for CM therapy, and the traditional medicine CuB as a candidate drug to hinder this pathway.

关键词： Conjunctival melanoma Cucurbitacin B Activity-based protein profiling G2/M cell cycle grp78 FOXM1 KIF20A Rare tumor

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Effect of aerobic exercise on grp78 and ATF6 expressions in mice with non-alcoholic fatty liver disease

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Sports Medicine and Health Science 2023年第2期5卷 112-119页

作者： Junhan Li Lei Huang Wei Xiong Chun Gu Shuling Zhang Xiali Xue School of Sports Medicine and Health Chengdu Sport UniversityChengduSichuan ProvinceChina Department of Exercise Rehabilitation Sichuan Provincial Orthopedic HospitalChengduSichuan ProvinceChina Department of Hepatobiliary Surgery Sichuan Provincial People's HospitalChengduSichuan ProvinceChina

Nonalcoholic fatty liver disease(NAFLD)is a prevalent medical condition with an ever-growing *** multiple intracellular mechanisms are involved,endoplasmic reticulum(ER)stress has been demonstrated to play a significant role in the genesis and *** of the research supports the advantages of exercise for ***,little is known about the molecular mechanism(s)that underpin the effectiveness of exercise training in *** study aimed to identify how aerobic exercise affected hepatic ER stress in a mouse NAFLD *** this study,the mice were fed either a standard diet(SD)or a high-fat diet(HFD)for 17 *** mice were trained on a treadmill during the last eight *** animals were tested for serum levels of biochemical assays,protein expression,and gene *** hematoxylin and eosin,Oil red O,and immunohistochemistry staining were also *** results indicated that a high-fat diet generated NAFLD,with serum lipid disruption and hepatic function impairment,and increased grp78 and ATF6 ***,aerobic training reversed the majority of these *** is concluded that NAFLD appears to be associated with hepatic ER stress response,and aerobic exercise mitigates NAFLD via lowering ER stress proteins grp78 and ATF6.

关键词： Endoplasmic reticulum stress Exercise grp78 ATF6 Nonalcoholic fatty liver disease

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grp78 upregulation-induced increase in cisplatin sensitivity of SPCA1 lung cancer cells

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Chinese Medical Journal 2011年第20期124卷 3341-3346页

作者： ZHANG Li-chuan WANG Jia-rui ZHAO Long WANG Tao WU Jing FAN Su-fang CHEN Li-xia SHAO Shu-juan Joseph Molnar WANG Qi Department of Respiratory Medicine Affiliated Zhongshan Hospital of Dalian University Dalian Liaoning 116001 China Graduate School of Dalian Medical University Dalian Liaoning 116044 China Department of Respiratory Medicine Fifth Hospital of Dalian Dalian Liaoning 116021 China Department of Respiratory Medicine Second Affiliated Hospital of Dalian Medical University Dalian Liaoning 116027 China Department of Respiratory Medicine Dalian No. 4 People＇s Hospital Dalian Liaoning 116031 China Department of Respiratory Medicine Second Hospital of Shijiazhuang Shijiazhuang Hebei 050051 China Department of Pediatrics Wuhan General Hospital of Guangzhou Military Region Wuhan Hubei 430070 China Department of Histology and Embryology Dalian Medical University Dalian Liaoning 116044 China Department of Medical Microbiology and Immunobiology Faculty of Medicine University of Szeged Dom ter 10 H-6720 Szeged Hungary

Background Glucose regulated protein 78 （grp78）, an endoplasmic reticulum （ER） chaperone, plays a critical role in chemotherapy resistance in a variety of cancers. In this study, we investigated the up-regulation of grp78 induced by A23187 and its association with the chemotherapeutical sensibility to cisplatin in human lung cancer cell line SPCAI. Methods SPCA1 cells were pretreated with A23187 at different concentrations. The expression of grp78 at the mRNA level was analyzed by RT-PCR; the expression of grp78 at the protein level was determined by Western blotting and immunofluorescence assay. Cell survival was determined by MTT assay. Cell apoptosis was analyzed by flow cytometry. Results The expression of grp78 at both the mRNA and protein levels was obviously induced by A23187 in SPCA1 cells, with an elevation of grp78 by 2.1-fold at the mRNA level and by 3.8-fold at the protein level compared to the control. There was a dose-dependent response. Survival curve analysis demonstrated that A23187 induction caused a significant reduction of survival for the cells subjected to cisplatin treatment （P 〈0.05）. After treatment by cisplatin, the percentage of apoptotic cells in the A23187 pretreated group increased about three fold compared with the control group （（27.53!-4.32）% vs. （9.25＋3.64）%, P 〈0.05）. Conclusions A23187 treatment was fairly effective for the induction of grp78 in SPCA1 cells at both the mRNA and protein levels. To a certain extent, grp78 up-regulation by A23187 was associated with the enhancement of drug sensitivity to cisplatin in human luncl cancer cell line SPCAI.

关键词： grp78 human lung cancer cell line SPCA1 cisplatin chemosensitivity

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葡萄糖调节蛋白grp78和grp94在小鼠脑发育过程中的差异表达

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生物化学与生物物理进展 2006年第7期33卷 641-646页

作者：李波王大鹏方文刚陈誉华中国医科大学发育生物学研究室卫生部细胞生物学重点实验室沈阳110001

为探讨葡萄糖调节蛋白grp78和grp94在小鼠脑发育过程中的生物学意义,利用蛋白质免疫印迹、免疫荧光及RNA印迹技术,检测了发育不同时期小鼠脑组织中grp78、grp94的表达及分布情况.结果显示:小鼠脑发育过程中grp78、grp94的表达在时间和... 详细信息

为探讨葡萄糖调节蛋白grp78和grp94在小鼠脑发育过程中的生物学意义,利用蛋白质免疫印迹、免疫荧光及RNA印迹技术,检测了发育不同时期小鼠脑组织中grp78、grp94的表达及分布情况.结果显示:小鼠脑发育过程中grp78、grp94的表达在时间和空间上呈现出显著差异,在脑发育的早中期grp78表达水平高于grp94,随发育的进程grp78不断下降而grp94逐渐升高,至胚胎发育晚期grp94表达水平高于grp78.在E16.5的不同脑区,grp78的表达呈现出从端脑到后脑逐渐递减的“浓度梯度”分布,而grp94在不同脑区中表达相同.小鼠出生后,二者作为应激蛋白在脑组织中的表达没有明显的差异性.免疫荧光结果显示,grp78和grp94在大脑组织中的分布基本相同,神经细胞和神经胶质细胞的细胞质均有分布.这些观察得到的结果提示,grp78和grp94与神经细胞分化和脑的形态建成有关,它们分别在脑发育的不同时期起作用.

关键词：葡萄糖调节蛋白 grp78 grp94 脑发育内质网分子伴侣

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Expression And Clinical Significance of Glucose Regulated Proteins grp78 And grp94 in Human Colon Cancer

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Chinese Journal of Cancer Research 2010年第1期22卷 42-48页

作者： Ming-hua Liu Meng-chunWang Na Gao Yan Li Wei-guo Jiang Department of Gastroenterology Department of pathology Shengjing Hospital China Medical University Shenyang 110004 China Endoscopy Center Liaoning Tumour Hospital Shenyang 110000 China

Objective： To investigate the expression of glucose regulated proteins grp78 and grp94 in human colon cancer. Methods： Tissues of resected primary colon cancer, colon adenoma and normal tissue were investigated. Protein expression was detected with immunohistochemical staining, mRNA expression levels of grp78 and grp94 were determined by semiquantitative reverse transcriptase polymerase chain reaction （RT-PCR） after mRNA extraction. Results： The expression of grp94 and grp78 was significantly higher in colon cancer when compared to those in colon adenoma and normal tissue （P〈0.01）. grp94 mRNA and protein expression was found to be in close relationship with the grade of differentiation, Dukes stages, lymph node involvement and remote metastasis in colon cancer （P〈0.01）, but no relationship with gender and age （P〉0.05）. grp78 mRNA and protein expression increased with cancer progression along the normal tissue-adenoma-cancer sequence, but showed no association with grade of differentiation, Dukes stages, lymph node involvement, remote metastasis, gender and age （P〉0.05）. The mRNA expression of grp78 and grp94 was consistent with the proteins （P〈0.01）, but there is no correlation between overexpression of grp78 and grp94 （P〉0.05）, and the patients with both strong grp78 and grp94 protein expression did not show advanced tumor stages （P〉0.05）. Conclusion： Overexpression of grp78 and grp94 was found in colon cancer. Overexpression of grp94 was closely related to cellular differentiation, Dukes stages, invasion and metastasis.

关键词： Colon cancer grp78 grp94 Colon adenoma Dukes stages Metastasis

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grp78表达对人肝细胞癌细胞外信号调节激酶1/2活性和表达的影响

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解剖学报 2009年第6期40卷 928-932页

作者：苏荣健李贞程留芳李宏丹魏嘉包翠芬辽宁医学院科学实验中心细胞和分子生物学与新药开发重点实验室辽宁锦州121001 解放军总医院消化科北京100583

目的通过检测葡萄糖调节蛋白78(grp78)、细胞外信号调节激酶(ERK1/2)、磷酸化ERK1/2在肝细胞癌手术切除组织中的表达,并在SMMC-7721细胞中干预grp78的表达,探讨grp78对ERK1/2激酶的调控作用。方法应用免疫组织化学方法(IHC)和免疫印迹... 详细信息

目的通过检测葡萄糖调节蛋白78(grp78)、细胞外信号调节激酶(ERK1/2)、磷酸化ERK1/2在肝细胞癌手术切除组织中的表达,并在SMMC-7721细胞中干预grp78的表达,探讨grp78对ERK1/2激酶的调控作用。方法应用免疫组织化学方法(IHC)和免疫印迹法检测47例肝细胞癌手术切除组织样本中grp78,ERK1/2的表达和ERK1/2磷酸化水平;在高表达grp78的肝细胞癌细胞系SMMC-7721中,通过用小RNA干扰grp78的表达来探讨grp78表达水平,改变对ERK1/2活性及表达的影响。结果在肝细胞癌组织样本中,grp78的表达情况与ERK1/2和磷酸化ERK1/2的表达明显呈正相关。在SMMC-7721细胞中grp78高表达促进ERK1/2的磷酸化,应用RNA干扰特异性下调grp78高表达细胞中grp78水平可以抑制ERK1/2的磷酸化。结论grp78参与调解ERK1/2信号通路,并且在肝细胞癌临床治疗方面可能成为潜在的治疗靶点。

关键词：肝细胞癌 grp78 细胞外信号调节激酶1/2 小RNA干扰 SMMC-7721 免疫组织化学免疫印迹法人

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盐度胁迫后青海湖裸鲤grp78及其相关基因的应答表达

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中国水产科学 2024年第1期31卷 70-81页

作者：赵静李长忠王蓉马淑兄贾春艳王璐勾化宇李兰英陈艳霞柳芳祁洪芳金文杰青海大学生态环境工程学院农业农村部高原冷水鱼养殖与生态环境保护重点实验室(部省共建) 青海湖裸鲤救护中心青海省青海湖裸鲤繁育与保护重点实验室

为深入研究青海湖裸鲤(Gymnocyprisprzewalskii)面临盐度胁迫时的分子应答机制,本研究克隆了青海湖裸鲤葡萄糖调节蛋白78 (glucose-regulated protein 78 kD, grp78)基因,并采用qPCR法检测5‰, 10‰和15‰盐度胁迫后grp78及其相关基因... 详细信息

为深入研究青海湖裸鲤(Gymnocyprisprzewalskii)面临盐度胁迫时的分子应答机制,本研究克隆了青海湖裸鲤葡萄糖调节蛋白78 (glucose-regulated protein 78 kD, grp78)基因,并采用qPCR法检测5‰, 10‰和15‰盐度胁迫后grp78及其相关基因的应答模式。结果显示,青海湖裸鲤grp78的开放阅读框长度为1962 bp,编码653个氨基酸,并包含HSP70超家族保守结构域。grp78在青海湖裸鲤9个组织中均有表达,其中在肠道、肝脏和心脏中的表达显著高于其他组织(n=3, Pgrp78、Hyou1、Prdx1、Nqo1和UBC基因的表达先被抑制,随后逐渐上调,而DNAJC2、Cu/Zn-SOD、Mn-SOD和Hmox1基因的表达则先上调后逐渐下调,随后又上调;在肾脏和肝脏中,DNAJC2、Cu/Zn-SOD、Mn-SOD、Hmox1和Nqo1基因的表达也呈现类似的先上调后下调再上调的模式。以上结果表明, grp78及其相关基因对盐度胁迫具有复杂的响应,可能参与了青海湖裸鲤适应和应对盐度胁迫的过程。此外, grp78及其相关基因Hyou1、DNAJC2、Hmox1、Nqo1、UBC、Prdx1、Cu/Zn-SOD和Mn-SOD参与了机体抗氧化应激调节,在减少盐度损伤方面发挥重要作用。

关键词：青海湖裸鲤盐度胁迫 grp78 基因克隆基因表达

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多巴胺诱导SH-SY5Y细胞帕金森病模型中grp78的表达

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吉林大学学报（医学版） 2010年第1期36卷 76-80,I0002页

作者：田明秀常明朱本清解洪荣王丹平张春光胡林森吉林大学第一医院神经内科吉林长春130021 天津市人民医院内科天津300130 武警吉林省总队医院骨科吉林长春130052

目的:探讨神经毒素多巴胺(DA)诱导SH-SY5Y细胞帕金森病(PD)细胞模型中葡萄糖调节蛋白78(grp78)的表达,为深入研究PD的发病机制提供理论依据。方法:建立多巴胺诱导SH-SY5Y细胞帕金森病模型,实验组加入DA(溶解于0.2%维生素C,DA终浓度为200... 详细信息

目的:探讨神经毒素多巴胺(DA)诱导SH-SY5Y细胞帕金森病(PD)细胞模型中葡萄糖调节蛋白78(grp78)的表达,为深入研究PD的发病机制提供理论依据。方法:建立多巴胺诱导SH-SY5Y细胞帕金森病模型,实验组加入DA(溶解于0.2%维生素C,DA终浓度为200μmol.L-1),对照组加入等体积的0.2%维生素C,分别孵育24h,台盼蓝染色检测细胞存活率,Hoechst33342染色检测细胞凋亡。分别提取实验组和对照组细胞总蛋白,应用荧光差异双向凝胶电泳(DIGE)技术获得蛋白点的差异表达,应用基质辅助激光解吸/电离飞行时间质谱仪(MALDI-TOFMS)鉴定差异蛋白质。结果:实验组细胞存活率为53.2%,与对照组细胞存活率(100%)比较显著下降(Pgrp78。结论:多巴胺能够诱导SH-SY5Y细胞凋亡,凋亡过程中grp78表达增高,提示grp78增高可能与PD的发病机制有关。

关键词： SH-SY5Y细胞多巴胺 grp78 荧光差异凝胶电泳

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肾气丸对Zucker糖尿病肥胖大鼠肾损伤、grp78及自噬相关因子的影响

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北京中医药大学学报 2023年第2期46卷 224-231页

作者：刘嘉鹏冯婧车宇娥张晓凤刘雨梦王谦李姝玉北京中医药大学中医学院北京102488

目的研究肾气丸对Zucker糖尿病肥胖(ZDF)大鼠肾脏损伤的影响及其可能机制。方法 20只雄性8周龄ZDF大鼠给予5C08高脂饲料诱导4周后,随意分为模型组与肾气丸组,7只同周龄雄性Zucker瘦型大鼠为对照组。肾气丸组大鼠每日灌服肾气丸水煎液(8... 详细信息

目的研究肾气丸对Zucker糖尿病肥胖(ZDF)大鼠肾脏损伤的影响及其可能机制。方法 20只雄性8周龄ZDF大鼠给予5C08高脂饲料诱导4周后,随意分为模型组与肾气丸组,7只同周龄雄性Zucker瘦型大鼠为对照组。肾气丸组大鼠每日灌服肾气丸水煎液(8.12 g/kg),模型组和对照组大鼠每日灌服等量生理盐水,连续给药4周。观察大鼠一般情况并检测空腹血糖(FBG)与24 h尿微量白蛋白(24 h U-mAlb);观察肾组织病理变化;实时荧光PCR和蛋白质印迹法分别检测肾组织grp78、Beclin-1和LC3的基因与蛋白表达。结果相比于模型组,肾气丸组大鼠表现出较好的一般状态,体质量增长率升高(Pgrp78基因和蛋白表达水平降低(P<0.05),Beclin-1蛋白表达水平降低(P<0.05),LC3蛋白表达水平升高(P<0.05)。结论肾气丸可能通过缓解内质网应激并促进自噬来减轻2型糖尿病动物ZDF大鼠的肾组织损伤,延缓糖尿病肾脏疾病进展。

关键词：肾气丸 2型糖尿病糖尿病肾脏疾病内质网应激自噬 grp78 Zucker糖尿病肥胖大鼠

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Chelation of grp78 with Lead and Its Localization Changes in the Astroglia of Rats Exposed to Lead

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2009年第4期29卷 492-497页

作者：张莹叶丽平王彪李妍孙黎光 Institute of Neurology China Medical University Department of Pathophysiology Liaoning Medical University Department of Biochemistry and Molecule Biology China Medical University

To observe the chelation of grp78 with lead （Pb） and its localization changes, astroglial ceils from Wistar rat brain were primarily cultured in medium witb acetate Pb. The processes were terminated at different time points. The immunoprecipitation （IP） and Western blotting were used for grp78 purification and expression and the Pb concentration was determined by employing atomic absorption spectrophotometry （AAS）. The localization change of grp78 was observed with colloid gold immunoelectron microscopy. The results showed that the expression of grp78 was increased significantly in the cells treated with 1.0 μmol/L acetate Pb for 24 h and peaked at 96-192 h （P〈0.01）, and at the 12th day, the expression of grp78 began to decrease but was still higher than normal （P〈0.05）. Pb content started to increase when cells were treated by acetate Pb for 24 h, and the peak appeared at 8 day （P〈0.01）, and then Pb content decreased gradually, but was still higher than normal （P〈0.05）. grp78 protein expression began to remarkably increase when it transferred from ER to the cytosol around the nuclei 24 h after treatment with Pb. It is concluded that grp78 in astroglia could strongly chelate with Pb ions and it might be a target protein of Pb.

关键词： lead astroglia grp78 chelation

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建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

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