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A new 2-aminosteroid induces cellular differentiation and upregulates the expression of MafB and egr-1 genes respectively in HL-60 and K562 leukemia cells

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Chinese Medical Journal 2005年第2期118卷 91-99页

作者： HEQun LIQiong YUANLin-bo HEJun ResearchLaboratoryofBloodPhysiology DepartmentofPhysiologyXiangyaSchoolofMedicineCentralSouthUniversityChangsha410078China

Background In previous work, we suggested that some 2aminosteroids inhibited proliferation and induced differentiation of both human and murine leukemia cells Here, we reported the actions of another new 2aminosteroid designated as H89712 on human leukemia cells Methods Cell colony counting and MTT assay were used to determine proliferation Cell morphology, histochemical staining, UV detection and cytometry were used to determine differentiation RTPCR was used to detect gene expression. Standard statistical method was used to analyze dataResults H89712 inhibited proliferation of HL60 leukemia cells and the inhibition percentage in MTT assay was 18% at the dose of 108 mol/L and 65% at the dose of 105 mol/L, respectively The inhibition for HL60 in colony assay was 23% at the dose of 108 mol/L and 96% at the dose of 105 mol/L, respectively H89712 also induced HL60 cells toward macrophagelike differentiation It was verified by flow cytometry that the percentage of positive CD14 expression in differentiated HL60 cells was about 9 times higher than that of the control at the dose of 108 mol/L and 20 times higher than that of the control at the dose of 105 mol/L respectively, and this action involved upregulation of MafB gene in HL60 leukemia cells On the other hand, H89712 inhibited proliferation of K562 leukemia cells and the inhibition of K562 leukemia cells in MTT assay was shown by 34% at the dose of 108 mol/L and 88% at the dose of 105 mol/L respectively The inhibition of K562 leukemia cells in colony assay was 53% at the dose of 108 mol/L and 100% at the dose of 105 mol/L respectively H89712 also induced K562 cells toward erythroidlike differentiation and it was verified by flow

关键词： 2-aminosteroid proliferation differentiation MafB egr-1 HL-60 K562 leukemia

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Ischemia/hypoxia inhibits cardiomyocyte autophagy and promotes apoptosis via the egr-1/Bim/Beclin-1 pathway

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Journal of Geriatric Cardiology 2020年第5期17卷 284-293页

作者： Bo SU Xian-Tao WANG Yu-Han SUN Man-Yun LONG Jing ZHENG Wen-Hao WU Lang LI Department of Cardiology the First Affiliated Hospital of Guangxi Medical UniversityNanningChina

Background Myocardial injury caused by microvascular obstruction(MVO)is characterized by persistent ischemia/hypoxia(IH)of cardiomyocytes after microembolization.Autophagy and egr-1 were closely associated with various cardiovascular diseases,including MVO.Bim and Beclin-1 are the important genes for autophagy and apoptosis.We aimed to explore whether the egr-1/Bim/Beclin-1 pathway is involved in regulating autophagy and apoptosis in IH-exposed cardiomyocytes.Methods Neonatal rat cardiomyocytes exposed to the IH environment in vitro were transfected with lentivirus expressing egr-1 or egr-1 sh RNA,or further treated with 3-methyladenine(3-MA).The expressions of autophagy and apoptosis-associated genes were evaluated using RT-q PCR and Western blots assays.Autophagic vacuoles and autophagic flux were detected by transmission electron microscopy(TEM)and confocal microscope,respectively.Cell injury was assessed by lactate dehydrogenase(LDH)leakage,and apoptosis was determined by flow cytometry.Results IH exposure elevated egr-1 and Bim expressions,and decreased Beclin-1 expression in rat cardiomyocytes.egr-1 overexpression in IH-exposed cardiomyocytes significantly up-regulated the levels of egr-1 and Bim,and down-regulated the level of Beclin-1.egr-1 knockdown resulted in down-regulated expressions of egr-1 and Bim,as well as up-regulated expression of Beclin-1.In addition,egr-1 knockdown induced autophagy was suppressed by 3-MA treatments.TEM and autophagic flux experiments also confirmed that egr-1 inhibited autophagy progression in IH-exposed cardiomyocytes.egr-1 suppression protected cardiomyocytes from IH-induced injury,as evidenced by the positive correlations between egr-1 expression and LDH leakage or apoptosis index in IH-exposed cardiomyocytes.Conclusions IH-induced cardiomyocyte autophagy and apoptosis are regulated by the egr-1/Bim/Beclin-1 pathway,which is a potential target for treating cardiomyocyte injury caused by MVO in the IH environment.

关键词： Autophagy Apoptosis Cardiomyocyte egr-1 Ischemia/hypoxia

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Early Growth Response gene 1 (egr-1) regulates HSV-1 ICP4 and ICP22 gene expression

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Cell Research 2007年第6期17卷 546-555页

作者： Gautam R Bedadala Rajeswara C Pinnoji Shao-Chung V Hsia Department of Basic Pharmaceutical Sciences College of Pharmacy The University of Louisiana at Monroe 700 University AvenueMonroeLA 71209 USA

The molecular mechanisms mediating herpes simplex virus type 1 (HSV-1) gene silencing during latent infection arenot clear. Five copies of early growth response gene 1 (egr-1) binding elements were identified in the intron of HSV-1ICP22 (infected cell protein No. 22) gene, leading to the hypothesis that egr-1 binds to the viral genome and regulates theviral gene expression. Transient co-transfection assays indicated that egr-1 negatively regulated the transcription of bothfull-length and intron-removed ICP22 promoters. The same assays also revealed that egr-1 repressed ICP4 (infected cellprotein No. 4) promoter activity in a dose-dependent manner but showed less inhibition when the intron was removed.Histone deacetylation was not involved in this regulation since histone deacetylase inhibitor trichostatin Adid not exhibitany effect on egr-1-mediated repression. Chromatin immunoprecipitation assays showed that egr-1 reduced the bindingof Spl to the promoters and that the co-repressor Nab2 (NGFI-A/egr1-binding protein) was recruited to the proximityof ICP4 in the presence of egr-1. These results suggested that the multifunctional transcription factor egr-1 can repressHSV-1 immediate-early gene expression through the recruitment of co-repressor Nab2 and reduction of Sp1 occupancy,and thus may play a critical role in HSV-1 gene silencing during latency.

关键词： HSV-1 latency egr-1 Sp1 Nab2 transcription regulation

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Inhibitory effect of 1,8-cineol (eucalyptol) on egr-1 expression in lipopolysaccharide-stimulated THP-1 cells

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Acta Pharmacologica Sinica 2007年第6期28卷 908-912页

作者： Jian-ya ZHOU~2,Xue-fen WANG~(2,4),Fa-di TANG~3,Jian-ying ZHOU~2,Guo-hua LU~2,Yan WANG~3,Ru-lian BIAN~3~2 Department of Respiratory Medicine,The First Affiliated Hospital,College of Medicine,Zhejiang University,Hangzhou 310003,China ~3 Zhejiang Respiratory Drugs Research Laboratory of State Drugs Administration of China,Zhejiang University,Hangzhou 310031,China Department of Respiratory Medicine The First Affiliated Hospital College of Medicine Zhejiang University Hangzhou China Zhejiang Respiratory Drugs Research Laboratory of State Drugs Administration of China Zhejiang University Hangzhou China

Aim:To study the effects of 1,8-cineol (eucalyptol) on the expression of earlygrowth response factor-1 (egr-1) and NF-r,B in the human monocyte THP-1 cellline stimulated by lipopolysaccharide (LPS).Methods:The THP-1 cells were incu-bated with serial doses of 1,8-cineol (1,10,and 100 mg/L,30 min) before beingstimulated with LPS (1 mg/L,30 min).The localization of egr-1 in the THP-1 cellswas detected by immunofluorescence and a laser scanning confocal microscope.The expression of egr-1 in the nuclei and whole cell,and NF-kB in the nuclei,weremeasured by Western blot analysis.Results:When stimulated by LPS,the FITC-labeled egr-1 was detected mainly in the nuclei.Moreover,the expression ofegr-1 in the whole cell increased markedly compared with the control cells.1,8-Cineol pretreatment decreased the expression of egr-1 in both the nuclei andwhole cell of the LPS-stimulated THP-1 cells,and this effect was concentration-dependent,but there was no reaction on the expression of NF-kB in the nucleiprotein in the LPS-stimulated THP-1 cells.Condusion:In a concentration-depen-dent manner,1,8-Cineol reduces LPS-induced egr-1 expression in nuclei and inwhole cell of THP-1 cells,but shows no effect on NF-kB expression.

关键词： 1,8-cineol egr-1 THP-lcells NF-kB lipopolysaccharide

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HIV-1 Protein Tat_(1–72) Impairs Neuronal Dendrites via Activation of PP1 and Regulation of the CREB/BDNF Pathway

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Virologica Sinica 2018年第3期33卷 261-269页

作者： Yu Liu Deyu Zhou Jiabin Feng Zhou Liu Yue Hu Chang Liu Xiaohong Kong Laboratory of Medical Molecular Virology School of Medicine Nankai University

Despite the success of combined antiretroviral therapy in recent years, the prevalence of human immunodeficiency virus(HIV)-associated neurocognitive disorders in people living with HIV-1 is increasing, significantly reducing the healthrelated quality of their lives. Although neurons cannot be infected by HIV-1, shed viral proteins such as transactivator of transcription(Tat) can cause dendritic damage. However, the detailed molecular mechanism of Tat-induced neuronal impairment remains unknown. In this study, we first showed that recombinant Tat(1–72 aa) induced neurotoxicity in primary cultured mouse neurons. Second, exposure to Tat_(1–72) was shown to reduce the length and number of dendrites in cultured neurons. Third, Tat_(1–72)(0–6 h) modulates protein phosphatase 1(PP1) expression and enhances its activity by decreasing the phosphorylation level of PP1 at Thr320. Finally, Tat_(1–72)(24 h) downregulates CREB activity and CREBmediated gene(BDNF, c-fos, egr-1) expression. Together, these findings suggest that Tat_(1–72) might impair cognitive function by regulating the activity of PP1 and the CREB/BDNF pathway.

关键词：蛋白质磷酸酶 HIV-1 损害小径树突激活 c-fos egr-1

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Targeting of p38 Mitogen-activated Protein Kinases to Early Growth Response gene 1 (egr-1) in the Human Paclitaxel-resistance Ovarian Carcinoma Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2008年第4期28卷 451-455页

作者：卢美松肖兰胡建莉邓锁徐艳 Department of Obstetrics and Gynecology the First Affiliated Hospital of Harbin Medical University Department of Obstetrics and Gynecology the Third Affiliated Hospital Sun Yat-sen University Center of Oncology Union Hospital Tongji Medical College Huazhong University of Science and Technology

To investigate the relationship between the expression of early growth response gene 1 （egr-1） and p38MAPK pathway in the paclitaxel resistance of ovarian carcinoma cells, the effect of p38MAPK inhibitor SB203580 on cell apoptosis was examined by using Hoechst 33258 staining. The intracellular Rh123 （Rhodamine 123） accumulation was detected by the flow cytometry （FCM）. The 50% inhibition concentration （IC50） of paclitaxel for A2780/Taxol cells was determined by MTT method. Electrophoretic motility shift assay （EMSA） was employed to examine the egr-1DNA binding activity. MDR1 and egr-1 mRNA were assessed by RT-PCR. The expressed of p-gp, phos- phorylated p53 and p38 were detected by Western blotting. SB203580 could remarkably promote the apoptosis of A2780/Taxol cells, and the cell apoptosis was in a time-dependent manner. Cellular Rh123 accumulation was increased, and the IC50 of paclitaxel for A2780/Taxol cells was decreased significantly. A2780/Taxol cell line after SB203580 treatment was shown to have a significantly higher level of egr-1 DNA binding activity. SB203580 down-regulated the activity of p38MAPK pathway, but up-regulated egr-1 expression. SB203580 significantly increased the level of cellular phosphorylated p53 protein, but decreased the p-gp protein level and MDR1 mRNA level in A2780/Taxol cells. There existed a close relationship between p38MAPK pathway and the paclitaxel resistance of ovarian carcinoma cells. The expression of egr-1 mediated by p38MAPK pathway plays a critical role in paclitaxel resistance of ovarian carcinoma cells.

关键词： ovarian carcinoma p38MAPK pathway egr-1 paclitaxel resistance

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葆青颗粒对环磷酰胺致早发性卵巢功能不全大鼠性激素及INHB、GDF-9、egr-1的影响

葆青颗粒对环磷酰胺致早发性卵巢功能不全大鼠性激素及INHB、GDF-...

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作者：张米佳西南医科大学

学位级别：硕士

目的:通过观察葆青颗粒对环磷酰胺致早发性卵巢功能不全大鼠性激素水平和GDF-9、egr-1蛋白表达的影响,探讨其作用及可能的作用机制.方法:选用50只动情周期正常的12周龄雌性SD大鼠,使用随机数字表随机分出10只空白组大鼠,除10只空白组大... 详细信息

目的:通过观察葆青颗粒对环磷酰胺致早发性卵巢功能不全大鼠性激素水平和GDF-9、egr-1蛋白表达的影响,探讨其作用及可能的作用机制.方法:选用50只动情周期正常的12周龄雌性SD大鼠,使用随机数字表随机分出10只空白组大鼠,除10只空白组大鼠外（注射同剂量氯化钠溶液）,余下大鼠使用75mg/kg CTX单次腹腔注射建立POI大鼠模型,造模后行阴道脱落细胞学确定造模成功,造模成功后,再次使用随机数字表将造模成功大鼠随机分为模型组、补佳乐组、葆青颗粒低剂量组、葆青颗粒高剂量组。模型组及葆青颗粒低剂量组各9只大鼠,余下各组各10只大鼠。补佳乐组予以戊酸雌二醇片0.21mg/kg/d灌胃,葆青颗粒低、高剂量组分别予以葆青颗粒3g/kg/d和9g/kg/d剂量灌胃,空白组与模型组予以同等容积氯化钠溶液灌胃,观察灌药后大鼠一般情况及动情周期变化,连续灌胃3周后,摘取双侧卵巢组织,HE染色观察卵巢组织形态学变化;用ELISA法检测大鼠血清FSH、LH、E2、INHB水平变化;用免疫组化法测定大鼠卵巢组织中GDF-9、egr-1蛋白表达水平变化。结果:造模3天及8天分别死亡大鼠1只,模型组及补佳乐组于灌胃第1周内各意外死亡大鼠1只及2只,空白组大鼠麻醉死亡3只,共有45只大鼠被收集卵巢标本,42只大鼠被收集血液标本。1.大鼠一般情况:1)空白组:大鼠精神良好,饮食、活动正常,毛发光亮,二便正常,体重明显增长（P＜0.05）。2)模型组:大鼠精神萎顿,不思饮食,喜扎堆、喜倦卧,毛发稀疏晦暗、脱毛,大便稀、不成形,偶有尿血,造模3周后,与空白组相比,体重增长速度缓慢（P＜0.05）。3)补佳乐组:随实验的进展,灌胃次数的增多,大鼠精神及饮食较前略有改善,脱毛改善不明显,二便正常,体重起始增长缓慢,随给药时间延长,体重增长较模型组更多,但无统计学差异（P＞0.05）。4)葆青颗粒低剂量组:随实验的进展,灌胃次数的增多,大鼠精神略有变好,饮食及脱毛改善不明显,二便正常,给药后体重略有增长,但与模型组差距不大,无统计学差异（P＞0.05）,且灌胃后2w及3w体重与同一时期的空白组比较,明显下降（P＜0.05）。5)葆青颗粒高剂量组:随着给药次数增多,大鼠精神及饮食较前略有改善,脱毛改善不明显,部分大鼠大便色黑、不成形,小便正常,体重起始增长缓慢,随给药时间延长,体重增长较模型组、葆青颗粒低剂量组更多,但无统计学差异（P＞0.05）,与补佳乐组相比,体重无明显差异（P＞0.05）。2.HE染色观察卵巢组织形态学变化:1)空白组:大鼠卵巢内可见始基、初级、次级、窦前、窦早期、排卵前卵泡等各级卵泡形态,数量充足,卵母细胞轮廓规整,颗粒细胞布列井然清晰,卵泡体大,泡液充裕,闭锁卵泡寥寥可数,卵巢髓质及皮质清晰可辨。2)模型组:大鼠卵巢结构欠明晰,各级卵泡数量较少,卵泡较小,卵泡液较少,空泡及闭锁卵泡数目彰着添长,未见优势卵泡。3)补佳乐组:卵巢结构清楚明了,各级卵泡了然可见,数量较模型组及葆青颗粒低剂量组明显增加,卵母细胞轮廓规整,颗粒细胞布列井然清晰,卵泡体大,泡液充裕,类似空白组.4)葆青颗粒低剂量组:各级卵泡了然可见,数量较模型组明显增加,可见成熟卵泡。但较空白组、补佳乐组及葆青颗粒高剂量组,卵泡数.目较少,整体卵泡较小,泡液较少。5)葆青颗粒高剂量组:卵巢结构清楚分明,各级卵泡了然可见,数量较模型组及低剂量组明显增加,卵母细胞轮廓规整,颗粒细胞布列井然清晰,卵泡形大,泡液充盈丰满,类似空白组及补佳乐组。3.ELISA法检测大鼠血清FSH、LH、E2、INHB水平:1)模型组:与其余四组相比,FSH、LH水平明显升高（P≤0.0001）,E2、INHB明显下降（除与葆青颗粒低剂量组E2相比:P≤0.01,余P≤0.0001）。2)补佳乐组:与空白组相比,FSH、LH水平升高,E2水平下降但无统计学意义（P＞0.05）,INHB下降明显（P＜0.05）;与葆青颗粒低剂量组相比,FSH、INHB略有下降,但无统计学意义（P＞0.05）,E2明显上升（P＜0.05）;与葆青颗粒高剂量组相比,FSH、LH无明显差异（P＞0.05）,E2略有上升,INHB略有下降,但无统计学意义（P＞0.05）。3)葆青颗粒低剂量组:与空白组相比,FSH、LH水平明显升高（FSH:P≤0.001;LH:P≤0.0001）,E2明显下降（P＜0.05）,INHB下降,但无统计学意义（P＞0.05）;与葆青颗粒高剂量组相比,FSH略高（P＞0.05）、LH明显升高（P≤0.0001）,E2略有下降（P＞0.05）,INHB无明显差异（P＞0.05）。4)葆青颗粒高剂量组:与空白组相比,FSH水平明显升高（P＜0.05）,LH 升高不明显（P＞0,05）,E2、INHB 略有下降（P＞0.05）。4.免疫组化法测定大鼠卵巢组织中GDF-9、egr-1蛋白表达水平:1)模型组:与

关键词：葆青颗粒环磷酰胺早发性卵巢功能不全性激素 GDF-9 egr-1

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A new 2-aminosteroid induces cellular differentiation and upregulates the expression of MafB and egr-1 genes respectively in HL-60 and K562 leukemia cells

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中华医学杂志(英文版) 2005年第2期 5-13页

作者： HE Qun, LI Qiong, YUAN Lin bo and HE Jun Research Laboratory of Blood Physiology, Department of Physiology, Xiangya School of Medicine, Central South University, Changsha 410078, China

关键词： 2-aminosteroid · proliferation differentiation MafB egr-1 HL-60 K562 leukemia

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Expression of egr-1,c-fos and cyclin D1 in esophageal cancer and its precursors:An immunohistochernical and in situ hybridization study

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World Journal of Gastroenterology 2004年第4期10卷 476-480页

作者： Ming-YaoWu Chu-XiangZhuang Huan-XingYang Ying-RuiLiang DepartmentofPathology ShantouUniversityMedicalCollegeShantou515031GuangdongProvinceChina DepartmentofPhysiology ShantouUniversityMedicalCollegeShantou515031GuangdongProvinceChina DepartmentofPathology TheFirstPeople'sHospitalofFoshanFoshan528000GuangdongProvinceChina

AIM:To examine the expression of egr-1, c-fos and cyclin D1 at both transcript and protein levels in esophageal carcinoma and to correlate the level of their expressions with precancerous and paracancerous esophageal lesions and esophageal carcinoma.METHODS:In situ hybridization and immunohistochemistry were used respectively to detect the expression of mRNA and proteins of egr-1, c-fos and cyclin D1 in 70 cases of esophageal squamous cell carcinoma and their corresponding para-cancerous mucosa and upper cut edge mucosa.RESULTS:In situ hybridization and immunohistochemistry showed positive staining of all three mRNAs in the cytoplasm and those of the proteins in nuclei. Overexpression of egr-1, c-los and cyclin D1 mRNAs and their proteins was found in dysplasia and squamous carcinomas. The expression level of egr-1 and c-los was high, and cyclin D1 was low in dysplasia mucosa, whereas the expression of egr-1 was decreased, c-fos was maintained and cyclin D1 was increased in the cancers. The expression of both c-fos and cyclinD1 was consistent between the mRNA and protein in their corresponding high expression lesions.CONCLUSION: The expression of egr-1, c-fos and cyclin D1 varies in esophageal precancerous lesions and cancer tissues, suggesting an involvement of these genes in the development of esophageal carcinoma.

关键词：食管癌细胞因子免疫组织化学癌前病变 egr-1 c-fos cyclin D1

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Relationship between egr-1 gene expression and apoptosis in esophageal carcinoma and precancerous lesions

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World Journal of Gastroenterology 2002年第6期8卷 971-975页

作者： Ming-Yao Wu Ying-Rui Liang Xian-Ying Wu,Department of Pathology,Shantou university Medical College,Shantou 515031,Guangdong Province,China Chu-Xiang Zhuang,Department of physiology,Shantou university Medical College,Shantou 515031,Guangdong Province,China Department of Pathology Shantou University Medical College Guangdong Province China. mywu@***

AIM: To study the expression of early growth response gene1 (egr-1 gene) and Bcl-X/L protein and its relationship with the cell apoptosis in human esophageal carcinoma(EC) and precancerous lesions.METHODS: In situ hybridization(ISH), immunohistochemistry (IHC) and TUNEL method were used respectively to detect egr-1mRNA, egr-1 protein, apoptosis related-protein Bcl-X/L and cell apoptosis in situ from 66 cases of esophageal squamous cell carcinoma and their upper cut edge and paracancerous mucosa.RESULTS: egr-1 gene in situ hybridization, Bcl-X/L immunohistochemistry positive products were located in the cytoplasm, while egr-1 immunohistochemistry and TUNEL positive signal were located in the nuclei. The apoptosis index(AI) and the frequency of apoptosis occurrence were increased gradually from precancerous lesion to cancer (P＜0.01) and the expression of egr-1mRNA and egr-1 protein in dysplasia was the highest among all specimens (P＜0.01).The AI of egr-1 positive cancer tissues was much higher than that of egr-1 negative cancer tissues (P＜0.01), while the AI of Bcl-X/L positive cancer tissues was much lower than that of Bcl-X/L negative cancer tissues (P＜0.01). The AI and egr-1 expression were not correlated with invasiveness and lymphatic metastasis in EC.CONCLUSION: Cell apoptosis was present through esophageal carcinogenesis. The expression of egr-1 mRNA and egr-1 protein were high in precancerous lesion of esophagus. The AI was increased significantly in egr-1 positive squamous cell carcinoma. egr-1 might promote apoptotic effect. egr-1 expression and cell apoptosis may have an important biological significance in esophageal carcinogenesis.

关键词：食管癌 egr-1 基因表达癌细胞凋亡癌前病变相关性

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