Study on the changes of nuclear matrix proteins during the differentiation of the human neuroblastoma SK-N-SH cells induced by retinoic acid

本论文以维甲酸（retinoicacid，RA）处理人神经母细胞瘤SK-N-SH细胞，鉴定RA对人神经母细胞瘤细胞的终末分化诱导效果。在此基础上，综合应用亚细胞蛋白质组学分析、免疫细胞化学、细胞分子生物学等技术，对在SK-N-SH细胞诱导分化过程中核基质蛋白的变化进行了较为系统的研究。分析鉴定和确证了与神经母细胞瘤细胞增殖分化相关的特异核基质蛋白，并研究特异核基质蛋白与人神经母细胞瘤细胞相关癌基因、抑癌基因在细胞内的共定位关系与变化，探索它们在神经母细胞瘤细胞诱导分化过程中的调控作用，以期能够在较为整体水平上进一步认识神经母细胞瘤细胞癌变与逆转机理问题，从而找出神经细胞增殖与分化相关的靶向性蛋...In this study, 1 μmol/L Retinoic Acid (RA) was used to induce the terminal differentiation of human neuroblastoma SK-N-SH cells, and its effects were investigated. On this base, by use of the methods of subcellular proteomics, immunocytochemistry and cellular molecular biology, we studied systematically the changes of nuclear matrix proteins during differentiation of SK-N-SH cells induced by RA, a...学位：理学博士院系专业：生命科学学院生物医学科学系_细胞生物学学号：2012005140313

An investigation of enhancing Chinese reading ability of junior secondary students by supplementation of film and television arts = 運用影視作品提升初中學生理解中文篇章的閱讀能力

Thesis (B. Ed.)--University of Hong Kong, 2013.指導老師: 羅燕琴.Zhi dao lao shi: Luo Yanqin.Includes bibliographical references (p. 31-33)published_or_final_versionB.Ed. in Language Educatio

Regulation of bioactive peptides of oyster(BPO)on the cell cycle and gene expression of human gastric adenocarcinoma cell-line BGC-823

目的比较研究牡蛎天然活性肽(BPO)对人胃腺癌BGC-823细胞凋亡的生物学效应,并进一步探索其对胃癌细胞的作用机制。方法采用酸抽提、凝胶柱层析等方法,从牡蛎体内分离提取到牡蛎天然活性多肽组分BPO-1,以姜黄素和HMBA处理组为平行对照,流式细胞仪检测细胞周期变化及以免疫细胞化学方法检测相关癌基因、抑癌基因表达变化。结果BPO-1能有效抑制BGC-823细胞增殖活动,出现亚G1期细胞,细胞进入凋亡现象。BPO-1,姜黄素及其组合均能不同程度地上调BGC-823细胞p16,c-myc,Fas,p21WAF1/CIP1蛋白表达,下调突变型p53,bcl-2蛋白表达。结论BPO-1对胃癌细胞具有显著的诱导凋亡作用。其诱导癌细胞凋亡机制与其调节和干预bcl-2,c-myc等癌基因和p53,p16,p21WAF1/CIP1等抑癌基因的表达有关。Objective To investigate the effects and mechanism of bioactive peptides of oyster(BPO)on the apoptosis of human gastric adenocarcinoma cell line BGC-823.Methods BPO-1,isolated from Ostrea cucullata by acid extraction and gel chromatography,was used to treat the BGC-823 cells.Curcumin and HMBA were used as control.Cell-cycle was detected by flow cytometer,some oncogene and tumor repressive gene were detected by lmmunocytochemistry analysis.Results BPO-1 could effectively inhibit the proliferation of BGC-823 cells,induce cells into sub-G1 phase.BPO1,curcumin and their combination could up-regulate the expression of gene p16,c-myc,Fas,p21WAF1/CIP1,and down-regulate gene p53,bcl-2.Conclusion BPO-1 could effectively induce BGC-823 cells into apoptosis,and the apoptosis effect involved to the changes of oncogene and tumor repressive gene.福建省科技计划重点资助项目(2003N0052

Effects of oyster bioactive peptides BPO-1 on the morphology and ultrastructure of human gastric adenocarcinoma cell line BGC-823

目的观察BPO-1对人胃腺癌BGC-823细胞形态与超微结构的影响,以探索BPO-1对胃癌细胞的作用机制。方法采用酸抽提、凝胶柱层析等方法,从牡蛎体内分离提取到牡蛎天然活性多肽组分BPO-1,以细胞凋亡诱导物姜黄素和癌细胞分化诱导物HMBA为平行对照,用光镜、透射电镜观察BPO-1处理细胞的形态和超微结构变化。结果经处理后的BGC-823细胞体积缩小、染色质凝聚、线粒体空泡化、内质网腔扩大和出现凋亡小体等多种典型的细胞凋亡形态和超微结构特征。结论牡蛎天然活性肽BPO-1对人胃腺癌BGC-823细胞凋亡的诱导具有显著作用。Objective To study the effects of bioactive peptides from oyster on the morphology and ultrastructure changes of human gastric adenocarcinoma cell line BGC-823 and its mechanism.Methods BPO-1 was isolated from Ostrea cucullata by acid extraction and gel chromatography,and was used to treat the BGC-823 cells.Curcumin and HMBA were used as control.The morphology and ultrastructure of the cells were examined by light microscopy and transmission electron microscopy.Results After treatment with BPO-1,typical apoptosis characteristics of the morphology and ultrastructure could be observed on BGC-823 cells.Conclusion BPO-1 could effectively inhibit the proliferation of BGC-823 cells and induce BGC-823 cells to apoptosis.福建省科技计划重点资助项目(2003N0052

EFFECTS OF TACHYPLESIN AND N-SODIUM BUTYRATE ON THE MORPHOLOGY AND ULTRASTRUCTURE CHANGES OF HUMAN GASTRIC ADENOCARCINOMA CELLS LINE BGC-823

目的以正丁酸钠为平行对照,比较观察鲎素和癌细胞诱导分化物对人胃癌细胞形态与超微结构变化的影响。方法采用光镜、扫描电镜与透射电镜,观察经2.0 mg/L鲎素、2.0 mmol/L正丁酸钠和1.0 mg/L鲎素+1.0 mmol/L正丁酸钠组合处理的人胃腺癌BGC-823细胞。结果光镜观察显示,鲎素和正丁酸钠以及鲎素+正丁酸钠处理的BGC-823细胞形态均较为一致,细胞体积增大、趋于扁平铺展状态,细胞核质比例减小,细胞核形状较圆整,核仁数量减少。扫描与透射电镜观察可见经鲎素、正丁酸钠及其组合处理的BGC-823细胞均出现细胞表面微绒毛稀少,细胞边缘丝状伪足减少,片状伪足增多;细胞核形态较规则,核内异染色质减少,常染色质增多:细胞质内线粒体增多,结构较为一致,高尔基复合体呈较为典型发达状态,并出现粗面内质网增多和多聚核糖体减少等变化。结论鲎素具有与正丁酸钠等诱导分化物相似的改变人胃癌细胞形态与超微结构恶性表型特征的作用,和与诱导分化物协同加成的诱导分化效果。Objective To compare the morphology and ultrastructure changes of human gastric adenocarcinoma cell line treated by tachyplesin and n-sodium butyrate. Methods Light microscope,scanning electron microscope and transmission electron microscope were used to observe human gastric adenocarcinoma cell line BGC-823 treated by 2.0?mg/L tachyplesin,2.0?mmol/L n-sodium butyrate and 1.0?mg/L tachyplesin in combination with 1.0?mmol/L n-sodium butyrate respectively. Results Light microscope observation showed that BGC-823 cells treated by tachyplesin,n-sodium butyrate and tachyplesin+n-sodium butyrate possessed the similar cell modality as follow: the volume of cell increased,the shape of cell became flat and outspread,nucleo-cytoplasmic ratio decreased,the shape of nucleus was rounde,the number of nucleolus decreased.Scanning electron microscope and transmission electron microscope observation showed that,in the BGC-823 cells which were treated with tachyplesin,n-sodium butyrate and tachyplesin in combination with n-sodium butyrate respectively,microvilli and filopodia reduced,sheed pseudopodia increased,the shape of nucleus became regular,heterochromatin decreased while euchromatin increased,the number of mitochondria increased and its structure appeared consistent,Golgi complex turned to be typical,the amount of rough endoplasmic reticulum increased and that of polyribosome decreased.Conclusion All of these results showed that tachyplesin possessed the similar effects to n-sodium butyrate on changing morphology and ultrastructure of human gastric adenocarcinoma cells and possessed an additive role of inducing tumor cells to differentiate cooperatively with n-sodium butyrate to induce carcinoma cell differentiation.国家自然科学基金资助[30170724

Differentiation Induced by Ginsenoside Rg1 Combined Formula Involved the Alteration of Prohibitin Localization and Expression in Human Osteosarcoma MG-63 Cells

选择性抽提经人参皂苷Rg1组合(RCT)诱导处理前后的人成骨肉瘤MG-63细胞核基质,对prohibitin在核基质中的存在、分布及其与相关基因产物在RCT处理前后MG-63细胞中的共定位关系进行观察研究.蛋白质组学分析结果显示,prohibitin存在于人成骨肉瘤MG-63细胞核基质蛋白组分中,并在RCT处理后细胞核基质中表达下调;蛋白质印迹杂交确证了prohibitin在MG-63细胞核基质中的存在及其在RCT处理后下调变化;免疫荧光显微镜观察进一步证实prohibitin定位在核基质上,经RCT处理后出现分布位置与表达水平变化;激光共聚焦显微镜观察可见prohibitin与c-Fos、c-Myc、p53和Rb基因产物均存在共定位关系,并在RCT处理后共定位分布区域出现变化.本研究证实了prohibitin是一种新发现的核基质蛋白,其在核基质上的定位与表达在RCT诱导分化前后发生显著变化,并与相关癌基因、抑癌基因产物存在共定位关系.实验表明RCT处理引起的prohibitin的变化与人成骨肉瘤MG-63细胞的诱导分化与调控具有密切关系,为深入揭示RCT等中药有效成分诱导肿瘤细胞分化的机理提供了重要科学依据和深入探索的新方向.Prohibitin is a nuclear matrix protein and associates with various oncogene products.A combined formula with ginsenoside Rg1,cinnamic acid and tanshinoneⅡA(RCT)was used to treat human osteosarcoma MG-63 cells in this study.We found that the prohibitin expression in the nuclear matrix was of decreased following RCT treatment by Western blot.The colocalization of prohibitin with the products of oncogenes or tumor suppressor genes,including c-Fos,c-myc,p53 and Rb was evaluated by immunofluorescence staining and laser scanning confocal microscopy.We observed significant alternations of prohibiitin distribution and the degree of colocalization with the associated gene products after the cells exposed to RCT.The results suggested that prohibitin might be involved in the RCT-induced differentiation in MG-63 cells.Our findings may provide information to expand the applications of Traditional Chinese Medicine formulas in cancer-related studies.国家自然科学基金资助项目(No.30470877)~

The Effects of Neonatal Bovine Brain Bioactive Peptide NBBP-1 on Proliferation and Associated Gene Expression of Human Neuroblastoma SK-N-SH Cells

目的:应用新生牛牛脑活性肽nbbP-1处理人神经母细胞瘤Sk-n-SH细胞,观察nbbP-1对人神经母细胞瘤Sk-n-SH细胞增殖和相关基因表达的影响。方法:通过细胞计数、流式细胞仪、光学显微镜、免疫细胞化学方法检测细胞的变化。结果:经60μg/MlnbbP-1处理后,Sk-n-SH细胞生长抑制率高达90.09%,细胞周期被阻滞在g0/g1,细胞核裂解成多个,免疫细胞化学染色结果显示,经处理后bCl-2抗凋亡基因蛋白表达减弱,而P53、fAS、bAX等促凋亡基因蛋白表达增强。结论:新生牛牛脑活性肽nbbP-1对人神经母细胞瘤Sk-n-SH细胞凋亡具有显著的诱导作用,其诱导癌细胞凋亡的机理与其调节和干预癌基因bCl-2和P53、fAS、bAX等抑癌基因的表达有关。Objective:To study the effects of neonatal bovine bioactive brain peptides in peakⅠ(NBBP-1),with which we treated SK-N-SH cells,on the proliferation and gene expression of human neuroblastoma SK-N-SH cells.Methods:The changes of SK-N-SH cells induced by NBBP-1 were detected by means of cell count,flow cytometry analysis,light microscope and immunocytochemistry.Results:Treated with 60 μg/mL NBBP-1,the proliferation of SK-N-SH cells was inhibited effectively,and the rate was up to 90.09 %.Cell cycle was arrested in G1/S phase.Nucleus broke into several parts.The results of immunocytochemistry showed that expression of anti-apoptotic gene bcl-2 was down-regulated and expression of apoptotic genes p53,fas,bax were up-regulated.Conclusions:This study suggested that NBBP-1 could regulate the proliferation of human neuroblastoma SK-N-SH cells effectively,and the effect was related with the changes of oncogene and tumor-suppressor gene.福建省科技计划重点项目(2003N0052)资

2008-2015年江苏省阜宁县居民恶性肿瘤死亡与减寿分析

随着经济的不断发展,生活习惯的改变和老龄化的加剧,我国疾病谱已发生了重要变化,慢性非传染病,尤其是恶性肿瘤已经成为影响居民健康的重要原因。近年来我国恶性肿瘤死亡率持续上升,已经成为居民死亡的主要原因[1]。江苏省是全国恶性肿瘤高发省份之一,恶性肿瘤死亡率高于全国平均水平[2]。为探讨恶性肿瘤对江苏省阜宁县居民健康和减寿的影响,本文对2008-2015年江苏省阜宁县居民恶性肿瘤国家自然科学基金(U1405221);;福建省科技重大专项(2015Y0051

类雌激素毒性快速检测试剂盒的制备方法

本发明公开了类雌激素毒性快速检测试剂盒的制备方法,首先是卵黄蛋白原的诱导和分离纯化,鱼体腹腔注射乙炔基雌二醇,用离子交换柱分离纯化鱼血清,透析脱盐,制备其冻干粉；其次是卵黄蛋白原多克隆抗体的制备,溶解冻干粉,加入弗氏佐剂乳化,免疫大白兔,制备多克隆抗体；第三是生物素标记的卵黄蛋白原多克隆抗体的制备,亲和柱分离纯化抗体,加入生物素酰-N-羟基丁二酰亚胺酯反应,在PBS缓冲液中透析,加入甘油,冻存。本发明可灵敏、准确、方便的定量检测多种鲤科鱼类血液中、肝组织、肝细胞培养基及鱼体中的卵黄蛋白原

一种检测鲤科鱼类卵黄蛋白原试剂盒的制备方法

本发明公开了一种检测鲤科鱼类卵黄蛋白原试剂盒的制备方法,首先是卵黄蛋白原的诱导和分离纯化,配制乙炔基雌二醇溶液,腹腔注射入鱼体,诱导一段时间,取鱼血,离心获得血清,用离子交换柱分离纯化,透析脱盐,制备冻干粉；其次是卵黄蛋白原多克隆抗体的制备,称取冻干粉,溶于双蒸水中,加入弗氏佐剂乳化,免疫大白兔,制备多克隆抗体。本发明可灵敏、准确、方便的定量检测多种鲤科鱼类血液中、肝组织及鱼体中的卵黄蛋白原