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| 1 | Apoptosis of hepatoma cells SMMC-7721 induced by Ginkgo biloba seed polysaccharide显示文摘AIM: To study the apoptosis of hepatoma cells SMMC-7721induced by polysaccharide isolated from Ginkgo biloba seed.METHODS: Ginkgo biloba seed polysaccharide (GBSP) wasisolated by ethanol fractionation of Ginkgo biloba seed andpurified by Sephadex G-200 chromatography. The purity ofGBSP was verified by reaction with iodine-potassium iodideand ninhydrin and confirmed by UV spectrophotometer,cellulose acetate membrane electrophoresis and Sepharose4B gel filtration chromatography. The Scanning ElectronMicroscope (SEM) and Flow Cytometrv (FCM) were used toexamine the SMMC-7721 cells with and without GBSPtreatment at 500 mg/ml for 36 h.RESULTS: GBSP product obtained was of high purity withthe average molecular weight of 1.86 × 105. Quantitativeanalysis of SMMC-7721 cells in vitro with FCM showed thatthe percentages of G2-M cells without and with GBSPtreatment were 17.01±1.28 % and 11.77±1.50% (P<0.05),the debds ratio of the cells were 0.46±0.12 % and 0.06±0 .06 %(P<0.01), and the apoptosis ratio of cells was 3.84±0 .55 %and 9.13±1.48 %(P<0.01) respectively. Following GBSPtreatment, microvilli of SMMC-7721 cells appeared thinnerand the number of spherical cells increased markedly. Mostsignificantly, the apoptosis bodies were formed on andaround the spherical cells treated with GBSP.CONCLUSION: GBSP could potentially induce the apoptosisof SMMC-7721 cells. | Gui-Wen Yang Li-Guo An Department of Biology,Shandong Normal University,Jinan 250014,Shandong Province,China Qun Chen Department of Biology & Chemistry,Huainan Teachers College,Huainan 232001,Anhui Province,China | 2002 | World Journal of Gastroenterology2002，8，5： | 15 |
| 2 | HMBA诱导人成骨肉瘤MG-63细胞分化过程中hnRNP A2/B1的表达与定位显示文摘背景与目的:核基质蛋白的差异表达与细胞癌变和增殖分化调控关系密切。本研究观察了hnRNP A2/B1在诱导分化处理前后人成骨肉瘤MG-63细胞核基质中的存在和分布,及其与Actin、Prohibitin的共定位关系。方法:选择性抽提经环六亚甲基双乙酰胺(hexamethylene bisacetamide,HMBA)诱导处理前后的MG-63细胞核基质,并运用双向电泳、质谱分析、蛋白质印记杂交、免疫荧光、激光共聚焦等技术检测hnRNP A2/B1在核基质中的表达与定位变化,及其与相关蛋白的共定位关系。结果:双向电泳及蛋白质印迹杂交结果证实了hnRNP A2/B1存在于MG-63细胞核基质蛋白组分中,并在HMBA处理后细胞核基质中表达下调;免疫荧光显微镜观察显示hnRNP A2/B1定位在核基质上,经HMBA处理后hnRNP A2/B1表达减弱。激光共聚焦显微镜观察结果显示,hnRNP A2/B1与细胞核基质蛋白组分Actin、细胞增殖相关调控因子Prohibitin具有共定位关系,但在诱导处理后细胞内的共定位关系减弱。结论:hnRNP A2/B1在MG-63细胞诱导分化过程中的表达分布,及其与Actin、Prohibitin的共定位关系的改变对MG-63细胞分化具有重要影响,值得进一步探索和研究。 | 赵春红 李祺福 陈兰英 唐剑 宋建晔 谢忠 | 2008 | 癌症2008，27，7： | 6 |
| 3 | Alreration of nuclear matrix-intermediate filament system and differential expression of nuclear matrix proteins during human hepatocarcinoma cell differentiation显示文摘AIM:To investigate the association between the configurational and compositional changes of nuclear matrix and the differentiation of carcinoma cells. METHODS: Cells cultured with or without 5 x 10'3 mmol/L of hexamethylene bisacetamide (HMBA) on Nickel grids were treated by selective extraction and prepared for whole mount observation under electron microscopy. The samples were examined under transmission electron microscope. Nuclear matrix proteins were selectively extracted and subjected to subcellular proteomics study. The protein expression patterns were analyzed by PDQuest software. Spots of differentially expressed nuclear matrix proteins were excised and subjected to in situ digestion with trypsin. The peptides were analyzed by matrix-assisted laser-desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Data were submitted for database searching using Mascot tool (www.matrixscience.com). RESULTS: The nuclear matrix (NM) and intermediate filament (IF) in SMMC-7721 hepatocarcinoma cells were found relatively sparse and arranged irregularly. The nuclear lamina was non-uniform,and two kinds of filaments were not tightly connected. After induction for differentiation by HMBA,the NM-IF filaments were concentrated and distributed uniformly. The heterogeneous population of filaments,including highly branched utrathin filaments could also be seen in the regular meshwork. The connection between the two kinds of filaments and the relatively thin,condensed and sharply demarcated lamina composed of intermediate-sized filaments was relatively fastened. Meanwhile,21 NM proteins changed remarkably during SMMC-7721 cell differentiation. Four proteins,i.e. mutant Pyst1,hypothetical protein,nucleophosminl,and LBP were downregulated,whereas four other proteins,eIF6,p44 subunit,(3-tubulin,and SIN3B were upregulated with the last one,SR2/ASF found only in the differentiated SMMC-7721 cells. CONCLUSION: The induced differentiation of SMMC-7721 cells by HMBA is accompanied by the configurational changes of nuclear matrix-intermediate filament (NM-IF) system and the compositional changes of nuclear matrix protein expression. These changes may be important morphological or functional indications of the cancer cell reversion. | Jian Tang Jing-Wen Niu Dong-Hui Xu Zhi-Xing Li Qi-Fu Li Jin-An Chen | 2007 | World Journal of Gastroenterology2007，13，20： | 4 |
| 4 | 伪狂犬病病毒Ea株诱导牛肾细胞凋亡及核基质蛋白的变化显示文摘目的 研究伪狂犬病病毒(PrV)是否具有诱导组织培养细胞发生细胞凋亡的功能及探讨凋亡细胞核 基质蛋白表达的变化。 方法 利用锥虫蓝(台盼蓝)染色绘制PrV在不同细胞上的增殖曲线;荧光染色观察凋亡 细胞核的形态特征;琼脂糖凝胶电泳分析细胞染色体DNA的片段化;高分辨率双向电泳分析细胞凋亡前后核基质 蛋白的表达差异。 结果 膜通透性的DNA特异性结合染料Hoechst33342染色显示,PrV感染后36h,牛肾 (MDBK)细胞核染色质开始固缩、凝聚,细胞核碎裂;DNA提取及琼脂糖凝胶电泳分析表明,细胞染色体DNA发生 片段化,形成“DNA梯状”条带(DNAladder);尽管PrV可以诱导MDBK细胞发生细胞凋亡,但不能诱导IBRS 2,BHK 21及PK 15细胞发生凋亡。MDBK细胞发生凋亡前后核基质蛋白的表达发生改变,存在表达上调、下调、诱导表达 及表达遏制等情况。 结论 PrV诱导细胞发生细胞凋亡是其致细胞死亡的形式之一,间接反映了PrV与宿主细 胞间复杂的相互作用。细胞凋亡前后核基质蛋白表达存在的差异说明PrV致MDBK细胞发生凋亡是自身基因产 物或诱导或抑制宿主细胞基因表达的结果。 | 李祥敏 钱平 陈焕春 郭东春 徐卓菲 金梅林 | 2005 | 解剖学报2005，36，2： | 2 |
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