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Oxidative stress battles neuronal bcl-xl in a fight to the death

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Neural Regeneration Research 2021年第1期16卷 12-15页

作者： Han-A Park Katheryn Broman Elizabeth A.Jonas Department of Human Nutrition and Hospitality Management College of Human Environmental SciencesThe University of AlabamaTuscaloosaALUSA Department of Internal Medicine Section of EndocrinologyYale UniversityNew HavenCTUSA

bcl-xl is a pro-survival protein of the bcl2 family found in the mitochondrial ***-xl supports growth,development,and maturation of neurons,and it also prevents neuronal death during neurotoxic *** article reviews the mechanisms and upstream signaling that regulate the activity and abundance of *** team and others have reported that oxidative stress is a key regulator of intracellular bcl-xl balance in *** stress regulates synthesis,degradation,and activity of bcl-xl and therefore neuronal *** apoptosis,pro-apoptotic bcl2 proteins such as Bax and Bak translocate to and oligomerize in the mitochondrial *** of oligomers causes release of cytochrome c and activation of caspases that lead to neuronal ***-xl binds directly to pro-apoptotic bcl2 proteins to block apoptotic *** anti-apoptotic roles of bcl-xl have been well documented,an increasing number of studies in recent decades show that protein binding partners of bcl-xl are not limited to bcl2 ***-xl forms a complex with F1Fo ATP synthase,DJ-1,DRP1,IP3R,and the ryanodine *** proteins support physiological processes in neurons such as growth and development and prevent neuronal damage by regulating mitochondrial ATP production,synapse formation,synaptic vesicle recycling,neurotransmission,and calcium ***,under conditions of oxidative stress,bcl-xl undergoes proteolytic cleavage thus lowering the abundance of functional bcl-xl in ***,oxidative stress alters formation of bcl-xl-mediated multiprotein complexes by regulating post-translational ***,oxidative stress regulates transcription factors that target the bcl-x gene and alter accessibility of microRNA to mRNA influencing mRNA levels of *** this review,we discussed how bcl-xl supports the normal physiology of neurons,and how oxidative stress contributes to pathology by manipulating the dynamics of bcl

关键词： ATP bcl-xl F1Fo ATP synthase mitochondria neuroprotection oxidative stress

Interaction between Bax and bcl-xl proteins confirmed by partial acceptor photobleaching-based FRET imaging

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Journal of Innovative Optical Health Sciences 2020年第3期3卷 81-91页

作者： Fangfang Yang Mengyan Du Xiaoping Wang Tongsheng Chen MOE Key Laboratory of Laser Life Science and College of Biophotonics South China Normal University Guangzhou 510631 P.R.China Department of Pain Management the First Affiliated Hospital of Jinan University Guangzhou 510630 P.R.China

Exact interaction mechanism between Bax and bcl-xl,two key bcl-2 family proteins,is an interesting and controversial *** acceptor photobleaching-based quantitative fluores-cence resonance energy transfer(FRET)measurement,PbFRET,is a widely used FRET quantification method in living *** this report,we implemented pixel-to-pixel PbFRET imaging on a wide-field microscope to map the FRET efficiency(E)images of single living HepG2 cells co-expressing CFP-Bax and *** E value between CFP-Bax and YFP-bcl-xl was 4.59%in cytosol and 11.31%on mitochondria,conclusively indicating the direct interaction of the two proteins,and the interaction of the two proteins was strong on mitochondria and modest in cytosol.

关键词： Bax bcl-xl protein-protein interaction FRET imaging living cells

RNA interference by expression of short hairpin RNAs suppresses bcl-xl gene expression in nasopharyngeal carcinoma cells

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Acta Pharmacologica Sinica 2005年第2期26卷 228-234页

作者： Fang LIU~2,Cheng-wei HE,Yue-fei ZHANG~4,Ke-yuan ZHOU~3~2Institute of Biochemistry and Molecular Biology ~4Department of Otorhinolaryngology,The First Affiliated Hospital,Guangdong MedicalCollege,Zhanjiang 524023,China Institute of Biochemistry and Molecular Biology Guangdong Medical College Zhanjiang China Department of Otorhinolaryngology The First Affiliated Hospital Guangdong Medical College Zhanjiang China

Aim:To evaluate a new plasmid mediated RNA interference(RNAi)system andinvestigate whether knock-down of bcl-xl by short hairpin RNA(shRNA)caninduce apoptosis of human nasopharyngeal carcinoma(NPC)cell line CNE-2Z ***:The plasmid containing mU6 promoter was subcloned to yield thepmU6 plasmid,recombinant plasmid expressing shRNA targeting bcl-xl gene wasdesigned and constructed,and were co-transfected cells with green fluorescenceprotein expressing *** cytometry was used to evaluate transfectionefficiency,and RT-PCR and Western blot were applied to analyze bcl-xl mRNAand protein levels,***:The shRNA expressed by the recombi-nant plasmid efficiently suppressed bcl-xl gene expression and induced apoptosisof NPC cells in ***:The recombinant plasmid can sufficientlymediate RNAi in CNE-2Z cells,and knock-down of the bcl-xl expression by shRNAsignificantly induced apoptosis in CNE-2Z *** results suggest this newsystem,mediated RNAi can be used as a tool for the study of gene function andgene therapy.

关键词： RNAi plasmid gene transfection short hairpin RNA bcl-xl apoptosis nasopharyngeal carcinoma cells

Effect of bax,bcl-2 and bcl-xl on regulating apoptosis in tissues of normal liver and hepatocellular carcinoma

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World Journal of Gastroenterology 2002年第6期8卷 1059-1062页

作者： Xiao-Zhong Guo Xiao-Dong Shao Min-Pei Liu Jian-Hua Xu Li-Nan Ren Jia-Jun Zhao Hong-Yu Li Di Wang,Department of Gastroenterology,General Hospital of Shenyang Military Region,Shenyang 110016,Liaoning Province,China Department of Gastroenterology General Hospital of Shenyang Military Region Liaoning Province China.

AIM: To investigate the expression of bax, bcl-2 and bcl-xl mRNA in the tissues of normal liver and hepatocellular carcinoma (HCC), and analyze the relationship between the expression of bax, bcl-2 and bcl-xl mRNA and clinical parameters of HCC ***: The expression of bax, bcl-2 and bcl-xl mRNA of normal liver and HCC was measured by Northern blot. Statistical analyses were made by t test and correlation ***: A very low mRNA level was indicated at bax,bcl-2 and bcl-xl in the HCC tissues in contrast to the tissues of normal liver by Northern blot analysis. The analyses of mRNA level revealed that HCC tissues exhibited a mean 7.6-fold decrease in bax, 4.2-fold in bcl-2 and 3.5-fold in bcl-xl in comparison with normal control tissues, respectively. Positive correlation was found between bax and bcl-xl (r=0.7061,P＜0.01). There was no significance between the mRNA expression of these three genes and age, gender, tumor differentiation and tumor stage of HCC patients. CONCLUSION: The results are consistent with the fact that apoptosis rarely occurs in normal livers but increases in HCC, indicating that bcl-2 and bcl-xl may play a very important role in regulating the apoptosis of normal liver and HCC.

关键词： bax bcl-2 bcl-xl 凋亡介导因子正常肝组织肝细胞癌组织细胞凋亡调节作用

TRAIL-induced expression of uPA and IL-8 strongly enhanced by overexpression of TRAF2 and bcl-xl in pancreatic ductal adenocarcinoma cells

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Hepatobiliary & Pancreatic Diseases International 2013年第1期12卷 94-98页

作者： Dong-Hui Zhou Li-Na Yang Christian Rder Holger Kalthoff Anna Trauzold Department of Oncology First Affiliated HospitalZhejiang University School of Medicine Division of Molecular Oncology Institute of Experimental Cancer ResearchCCC-NorthUniversity Hospital Schleswig-HolsteinCampus Kiel24105 KielGermany

BACKGROUND:The death ligand,tumor necrosis factor(TNF)related apoptosis-inducing ligand(TRAIL),induces apoptosis and non-apoptotic signaling in some tumor *** purpose of this study was to investigate the roles of the pro-apoptotic TRAIL receptors,TRAIL-R1 and TRAIL-R2,as well as bcl-xl and TRAF2 in TRAIL-induced expression of the pro-inflammatory cytokine IL-8 and the invasion-promoting protein urokinase(uPA) in pancreatic ductal adenocarcinoma(PDAC) ***:Colo357wt,Colo357/TRAF2,Colo357/bcl-xl,Panc89 and PancTuI cells were stimulated with TRAIL and uPA and IL-8 expression was detected using real-time ***,receptor-specific antibodies were used to investigate the effects of TRAIL-R1 or TRAIL-R2 ***:Dose-dependent increases in uPA and IL-8 expression were detected following TRAIL stimulation in PDAC *** effects were inhibited when TRAIL-R1 but not TRAIL-R2 was *** of TRAF2 or bcl-xl strongly increased TRAIL-mediated upregulation of uPA and ***:In PDAC cells,TRAIL strongly induced uPA and IL-8 via *** response was further enhanced in cells overexpressing TRAF2 and ***,inhibition of the non-apoptotic "side-effects" of TRAIL treatments by inactivation of TRAF2 and bcl-xl might represent additional relevant strategies for the treatment of pancreatic cancer.

关键词： TRAIL uPA IL-8 bcl-xl pancreatic cancer cell

Cationic Liposome-mediated bcl-xl Gene Transfection into Human Keratocytes

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2005年第3期25卷 365-367页

作者：刘磊李新宇朱雪菲李贵刚 Department of Ophthalmology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology

The efficiency and safe range of LipofectamineTM2000 (LF2000)/bcl-xl applied in human keratocytes, the optimal ratio of LF2000/bcl-xl and the bcl-xl gene expression in human keratocytes were investiaged. By using trypan-blue staining, the effects of LF2000 and bcl-xl on the survival rate of the cultured human keratocytes were measured respectively. By using semi-quantitative RT-PCR, the efficiency and the expression of LF2000-mediated bcl-xl transfection into keratocytes were examined. The results showed that the survival rate of human keratocytes had no signficant change in the presence of LF2000 (20 μg/ml) or bcl-xl (10 μg/ml) for 24 h. LF2000 could effectively mediate the transfection of exogenous gene bcl-x1 into human keratocytes. The best transfection efficiency could be obtained when the ratio of bcl-xl/LF2000 was 1:8. One day after transfection, the positive cells for bcl-x1 could be detectable, and the positive rate reached the peak on the post-transfection day 3 (48.3 %), then gradually decreased. Fifteen days after transfection, there were few positive cells. It was suggested that LF2000 could effectively transfer the exogenous gene bcl-xl into human keratocytes without obvious toxicity during a concentration range. LF2000/bcl-xl may be likely to play an important role in gene therapy of human keratocytes.

关键词： bcl-xl stroma cells gene transfection cationic liposome

Silencing of bcl-xl Expression in Human MGC-803 Gastric Cancer Cells by siRNA

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Acta Biochimica et Biophysica Sinica 2005年第8期37卷 555-560页

作者： Xiao-Yong LEI~(#*) Miao ZHONG~# Lan-Fang FENG Chun-Yan YAN Bing-Yang ZHU Sheng-Song TANG Duan-Fang LIAO Institute of Pharmacy and Pharmacology,Nanhua University,Hengyang 421001,China InstituteofPharmacyandPharmacology NanhuaUniversityHengyang421001China

To investigate the inhibitory effect of the bcl-xl small interfering RNA(siRNA)on BcI-xlgene expression in the human gastric cancer cell line MGC-803,green fluorescent protein(GFP)siRNAwas constructed and transfected into MGC-803 ceils,together with GFP expression vector pTrace *** expression levels were observed using fluorescence ***-xl siRNA and negative siRNAwere then constructed and stably transfected into MGC-803 ***-PCR and immunofluorescence wereused to detect the expression of *** apoptosis was detected by acridine orange(AO)andflow *** were as follows:(1)48 h after GFP expression vector and GFP siRNA co-transfection,the expression level of GFP in the GFP siRNA group was much lower than the negative siRNA group,according to fluorescence microscopy *** mRNA and protein levels of bcl-xl in bcl-xl siRNAstable transfectants were reduced to almost background level compared with negative siRNA transfectantsor untreated cells.(2)Changes in nucleus morphology was observed by AO staining nucleic and flowcytometry analysis,which showed that stable bcl-xl siRNA transfectants have an increased spontaneousapoptosis (21.17%+1.26% vs.1.19%+0.18% and 1.56%+0.15% respectively,Pbcl-xl genes can specifically suppress GFP or BcI-xlexpression in MGC-803 cells,and bcl-xl siRNA can increase spontaneous ***-xl siRNA maybe a beneficial agent against human gastric adenocarcinoma.

关键词： bcl-xl siRNA MGC-803 cells

bcl-xl小分子干扰RNA逆转人结肠癌获得性肿瘤坏死因子相关凋亡诱导配体的耐药

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中华肿瘤杂志 2008年第4期30卷 245-249页

作者：朱洪波黄学锋胡静姿周玮陈薇陈琳琳何超浙江大学医学院附属邵逸夫医院肛肠外科生物医学研究中心杭州310016 南京军区杭州疗养院空勤疗养区

目的探讨bcl-xl小分子干扰RNA（siRNA）在逆转人结肠癌细胞获得性肿瘤坏死因子相关的凋亡诱导配体（TRAIL）耐药中的作用。方法以bcl-xl siRNA转染获得性TRAIL耐药的人结肠癌DLD1-TRAIL／R细胞24h，并用TRAIL蛋白处理，通过细胞计数和... 详细信息

目的探讨bcl-xl小分子干扰RNA（siRNA）在逆转人结肠癌细胞获得性肿瘤坏死因子相关的凋亡诱导配体（TRAIL）耐药中的作用。方法以bcl-xl siRNA转染获得性TRAIL耐药的人结肠癌DLD1-TRAIL／R细胞24h，并用TRAIL蛋白处理，通过细胞计数和流式细胞仪检测细胞的存活率，并通过Western blot法检测各种凋亡相关蛋白的活化情况。结果bcl-xl siRNA能有效下调DLD1-TRAIL／R细胞中bcl-xl蛋白的表达水平，并且能够有效地逆转其对TRAIL蛋白的耐药。DLDl-TRAIL／R细胞经过bcl-xlsiRNA和TRAIL蛋白共同处理2h后，其细胞凋亡率〉50％；共同处理4h后，其细胞存活率〈40％；而对照组和TRAIL蛋白处理组的细胞凋亡率和生存率无明显变化（P〈0．05）。Western blot法检测结果表明，经bcl-xl siRNA与TRAIL蛋白联合处理后，DLDl-TRAIL／R细胞中caspase-8、caspase-9、Bid、caspase-3以及多聚腺苷酸二磷酸核糖聚合酶（PARP）均明显活化，细胞色素C的释放显著增加。结论bcl-xl siRNA能够有效逆转结肠癌细胞获得性TRAIL耐药，这将为治疗肿瘤耐药提供一种新的思路。

关键词： bcl-xl 小分子干扰RNA 肿瘤坏死因子相关的凋亡诱导配体耐药

bcl-xl反义寡核苷酸增强食管癌细胞对5-氟尿嘧啶的敏感性

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中华肿瘤杂志 2006年第3期28卷 173-177页

作者：张蕾温洪涛王留兴张岚张云汉郑州大学第一附属医院肿瘤科河南省肿瘤病理重点实验室 450052 郑州大学第一附属医院消化内科 450052 郑州大学第一附属医院病理科 450052

目的探讨bcl -xl反义寡核苷酸(ASODN)在下调bcl- xl表达、增加食管癌细胞株EC9706对5氟尿嘧啶(5Fu)敏感性中的作用。方法设细胞对照组、空白对照组、无关序列寡核苷酸(N ODN)组、ASODN组、5Fu组及ASODN联合5Fu组,应用四甲基偶氮唑蓝(MTT... 详细信息

目的探讨bcl -xl反义寡核苷酸(ASODN)在下调bcl- xl表达、增加食管癌细胞株EC9706对5氟尿嘧啶(5Fu)敏感性中的作用。方法设细胞对照组、空白对照组、无关序列寡核苷酸(N ODN)组、ASODN组、5Fu组及ASODN联合5Fu组,应用四甲基偶氮唑蓝(MTT)法检测EC9706细胞的增殖抑制率;RT PCR和Westernblot检测bcl- xlmRNA和蛋白表达水平的改变;吖啶橙荧光染色和流式细胞技术定量检测EC9706细胞的凋亡率。结果bcl -xlASODN联合5Fu治疗组对食管癌细胞增殖的抑制率为71.58%,对bcl xlmRNA表达抑制率为81.25%,并可显著下调bcl xl的蛋白表达;凋亡指数为69.5%,凋亡率为(63.32±9.23)%,与细胞对照组、空白对照组、N -ODN组、ASODN组及5Fu组比较,差异均有统计学意义(Pbcl xl表达,可显著增强食管癌细胞对5Fu的敏感性。

关键词：食管肿瘤 5-氟尿嘧啶 bcl-xl 反义寡核苷酸