hS100A6对人骨肉瘤细胞株143B的体内体外作用

为研究hS100A6对人骨肉瘤细胞系143B的体内体外作用,利用整合有S100A6基因和SiS100A6基因的腺病毒(AdS100A6和AdSiS100A6)作用于143B细胞,MTT法和台盼蓝染色法检测其对细胞的增殖作用,Hoechst染色法检测其对凋亡的作用,Transwell实验检测其对迁移的作用,同时以143B裸鼠皮下移植瘤为动物模型,检测S100A6对143B的体内作用,结果显示AdS100A6干预后出现细胞增殖活性的减弱、凋亡率增高、迁移率降低,在体内试验中瘤体体积明显较对照组减小,而AdsiS100A6干预后出现时间依赖性地细胞增殖活性的增强、凋亡率降低、迁移率增高,体内试验中瘤体体积较对照组明显增大.提示hS100A6能抑制人骨肉瘤细胞株143B细胞的增殖和迁移,并促进细胞凋亡,同时能抑制人骨肉瘤细胞株143B的裸鼠皮下移植瘤的生长.     

葡萄籽原花青素对人骨肉瘤143B细胞的作用及机制研究

该文研究葡萄籽原花青素(grape seed procyanidins,GSP)对人骨肉瘤143B细胞增殖、凋亡的影响及其机制。用不同剂量GSP处理骨肉瘤143B细胞,结晶紫染色和集落形成试验分别用于检测细胞增殖和集落形成能力的变化;流式细胞术检测细胞周期与凋亡,Western blot法检测周期和凋亡相关蛋白;RT-PCR和Western blot法检测Wnt/β-catenin信号通路相关分子表达的变化;采用腺病毒Adβ-catenin感染以过表达β-catenin,探讨GSP抑制143B细胞增殖的作用是否与Wnt/β-catenin信号通路相关。结果显示,GSP在20~60μg/m L剂量范围内呈剂量和时间依赖性抑制143B细胞增殖活力和降低细胞克隆形成能力;GSP引起G0/G1周期阻滞和周期相关蛋白cyclin D1下调,还使细胞凋亡率明显增高,并伴有活化的凋亡相关蛋白cleaved PARP(poly ADP-ribose polymerase)和cleaved caspase-8的水平增高;GSP降低143B细胞中β-catenin m RNA水平及核内与总β-catenin蛋白质水平,抑制GSK3β磷酸化并上调GSK3β蛋白质水平;过表达β-catenin可部分减弱GSP对该细胞增殖活性的抑制作用。结果表明,GSP抑制骨肉瘤143B细胞的增殖和促进其凋亡,其机制涉及Wnt/β-catenin信号通路的抑制。     

S100A6蛋白对细胞中β-catenin水平的影响及可能机制

探讨S100A6蛋白对细胞中β-catenin水平的影响及可能机制。用表达S100A6及其siRNA的重组腺病毒AdS100A6和AdsiS100A6处理人骨肉瘤细胞系143B,Western blot分析处理前后细胞中β-catenin水平的变化;再以人胚肾细胞HEK293为研究对象,通过免疫共沉淀(co-immunoprecipitation,Co-IP)联合Western blot检测S100A6与Wnt经典信号通路主要成员β-catenin、糖原合酶激酶-3β(GSK-3β)、散乱蛋白(Dishevelled,DVL)和轴蛋白(Axin)之间的相互作用。结果:1)AdS100A6作用48h后,143B细胞中β-catenin水平较实验对照组(AdGFP组)增加了64%(P<0.01),而AdsiS100A6处理的细胞中β-catenin较AdGFP组减少了20.2%(P<0.05),AdGFP组与空白组之间的差异无统计学意义;2)在S100A6抗体的沉淀物中,检测到β-catenin、GSK-3β和DVL,而无Axin检出;3)β-catenin、GSK-3β和DVL的抗体沉淀物中,均有S100A6检出,而Axin抗体的沉淀物中无S100A6检出。结论:S100A6能够上调人骨肉瘤细胞系143B中的β-catenin水平;并与β-catenin、GSK-3β和DVL之间存在直接相互作用;S100A6与这三个分子之间的直接相互作用可能抑制了β-catenin的降解,使之在胞浆内聚集,水平升高。这可能是S100A6上调β-catenin的部分机制。     

DLX1过表达对骨肉瘤细胞MG63迁移、侵袭、凋亡和增殖的影响

前期经基因芯片筛选发现,同源盒基因1(distal-less homeobox 1,DLX1)低表达是导致骨肉瘤形成的关键基因。该研究在此基础上,拟通过过表达DLX1探讨其对骨肉瘤细胞MG63迁移和侵袭等生物学特征的影响,并初步揭示其作用途径。采用含DLX1基因的重组腺病毒Ad DLX1(adenovirus distal-less homeobox 1)和空载腺病毒Ad RFP(adenovirus red fl uorescence protein)分别感染人骨肉瘤细胞MG63,观察细胞荧光表达情况,并用RT-PCR和Western blot验证DLX1表达水平;Transwell实验检测细胞迁移和侵袭能力;DAPI染色和流式细胞术检测细胞凋亡水平;CCK-8检测细胞增殖能力;RT-PCR和Western blot检测Wnt/β-catenin信号通路中β-catenin的表达。结果显示,与对照组(Ad RFP感染组)相比,Ad DLX1能显著增加MG63细胞中DLX1的表达,并导致细胞迁移和侵袭能力下降,但细胞增殖和凋亡情况无明显改变。DLX1过表达还可以上调Wnt/β-catenin信号通路中β-catenin的表达。该研究表明,DLX1可通过Wnt/β-catenin信号通路抑制骨肉瘤细胞MG63的迁移和侵袭。     

OPN对卵巢癌Hey细胞增殖、迁移和侵袭能力的影响

目的:探讨骨桥蛋白(Osteopontin,OPN)对卵巢癌Hey细胞增殖、迁移和侵袭能力的影响及其可能涉及的分子机制。方法:选择卵巢癌细胞株Hey、HO8910、A2780和正常卵巢上皮细胞IOSE80,采用Western blot检测OPN蛋白的表达情况。对OPN表达量相对较高的Hey细胞株的OPN基因敲减或过表达,运用CCK-8、平板克隆实验、细胞划痕、Transwell侵袭实验等方法研究OPN对细胞增殖、迁移及侵袭能力的影响,采用Western blot检测Wnt/β-catenin通路相关蛋白β-catenin、CyclinD1、c-myc的表达情况。结果:OPN在卵巢癌细胞株Hey、HO8910、A2780内均有表达,且表达量均高于正常卵巢上皮细胞IOSE80。si RNA-OPN转染卵巢癌Hey细胞沉默OPN表达,CCK-8和平板克隆实验显示沉默OPN能够降低Hey细胞的增殖能力,划痕实验和Transwell侵袭实验显示下调OPN可降低Hey细胞的迁移和侵袭能力,Wnt/β-catenin通路相关蛋白β-catenin、CyclinD1、c-myc表达下降。ex-OPN转染Hey细胞使OPN表达升高,与对照组相比,OPN过表达能够增强Hey细胞的增殖、迁移和侵袭能力,并且Wnt/β-catenin通路相关蛋白β-catenin、CyclinD1、c-myc表达升高。结论:OPN能够促进卵巢癌细胞的增殖、迁移及侵袭能力,该作用可能通过促进Wnt/β-catenin信号通路实现。     

汉黄芩素抗骨肉瘤143B细胞的实验研究

目的:观察汉黄芩素对人骨肉瘤细胞系143B增殖和凋亡的影响,并探讨其可能的作用机制。方法:采用体外培养人成骨肉瘤细胞系143B,CCK-8实验检测不同浓度汉黄芩素对骨肉瘤143B细胞增殖抑制作用;流式细胞术分析汉黄芩素对癌细胞周期分布及凋亡的影响;Western Blot检测凋亡相关蛋白Bax、Bcl-2、cleaved caspase-9和cleaved caspase-3的表达水平。结果:CCK-8结果显示汉黄芩素以时间、浓度依耐性的方式抑制骨肉瘤143B细胞的增殖;流式细胞术结果表明汉黄芩素可导致骨肉瘤细胞周期阻滞于G0/G1期并以浓度依赖的方式诱导骨肉瘤细胞凋亡;Western Blot检测结果证明,汉黄芩素可上调骨肉瘤细胞中促凋亡蛋白Bax、cleaved caspase-9、cleaved caspase-3的表达,而下调抑制凋亡蛋白Bcl-2。结论:汉黄芩素抑制骨肉瘤细胞增殖、导致细胞周期阻滞,促进其凋亡,并呈现时间和浓度依赖性,汉黄芩素激活Caspase凋亡途径及诱导细胞周期阻滞可能是其抗骨肉瘤的作用机制。     

MicroRNA-21靶向调控β-catenin促进骨肉瘤细胞U2OS的增殖与侵袭

目的:研究微小RNA-21(microRNA-21,miR-21)对骨肉瘤细胞U2OS增殖与侵袭的影响及其可能机制。方法:采用Real-time PCR (RT-PCR)检测miR-21在骨肉瘤和临近正常骨组织中的表达差异。通过脂质体转染法将miR-21模拟物(microRNA-21mimics,即mimics组)及microRNA无关序列(microRNA-NC,即NC组)转染入骨肉瘤细胞U2OS,real-time PCR(RT-PCR)检测miR-21和β-catenin m RNA在U2OS细胞中的表达,Western blot检测β-catenin蛋白在U2OS细胞中的表达,并通过双荧光素酶报告基因验证miR-21与β-catenin基因3'-非编码区(3'-untranslated region,3'-UTR)的特异性结合作用。MTT法检测U2OS细胞体外增殖能力;Transwell侵袭模型探查U2OS细胞侵袭潜能。结果:骨肉瘤组织中miR-21水平显著高于正常骨组织(P0.05)。过表达miR-21能够增强细胞增殖与侵袭,上调U2OS细胞β-catenin m RNA和蛋白的表达。双荧光素酶报告基因结果表明miR-21可与β-catenin基因3'-UTR结合,从而对β-catenin的表达起调控作用。结论:miR-21可能通过调节β-catenin的表达促进骨肉瘤细胞U2OS的增殖与侵袭。     

白皮杉醇对子宫内膜癌HEC-1A细胞增殖、迁移和侵袭的影响

本研究旨在考察白皮杉醇(piceatannol, PIC)对子宫内膜癌细胞HEC-1A的初步抗肿瘤作用。分别通过CCK-8实验、平板克隆形成实验、流式细胞术、划痕损伤修复实验和Transwell小室实验检测不同浓度的PIC对HEC-1A细胞增殖、凋亡、迁移和侵袭能力的影响。Western blot实验检测细胞增殖相关蛋白和侵袭迁移相关蛋白的表达水平。结果显示，与空白组相比，PIC能显著抑制HEC-1A细胞增殖、克隆形成能力、诱导其凋亡并减少细胞迁移距离和侵袭细胞数(P<0.01),下调p-Akt、p-Erk1/2、p-p38MAPK、β-catenin、CD44和Slug蛋白的表达水平，上调E-cadherin蛋白的表达水平(P<0.01)。综上所述，PIC呈浓度依赖性抑制子宫内膜癌HEC-1A细胞的增殖、迁移与侵袭，其机制可能与抑制AKT/ERK/MAPK信号通路和影响Wnt/β-catenin信号通路和上皮-间质转化标志物的改变有关。     

甲基莲心碱促进骨肉瘤143B细胞凋亡的相关研究

目的:研究不同浓度甲基莲心碱对骨肉瘤143B细胞增值迁移的影响及其诱导凋亡的相关机制。方法:不同浓度甲基莲心碱处理骨肉瘤143B之后,CCK-8法检测甲基莲心碱对骨肉瘤143B细胞的增值抑制作用;细胞划痕实验检测不同浓度甲基莲心碱对骨肉瘤143B细胞迁移的影响;流式细胞术检测甲基莲心碱对癌细胞的凋亡率和周期分布;Western Blot检测甲基莲心碱对癌细胞相关蛋白Bcl-2、Bax的表达。结果:CCK-8显示甲基莲心碱能抑制骨肉瘤143B细胞的增值且以浓度和时间依赖的方式(P0.05);给药组(20, 40, 60μmol·L~(-1))24 h细胞迁移率分别为(62.35±4.15)%,(40.74±4.80)%,(25.10±5.52)%,较对照组(75.89±5.24)%明显降低(P0.05);流式细胞术结果显示甲基莲心碱能使骨肉瘤143B细胞周期阻滞于G0/G1期,且呈剂量依赖性诱导癌细胞凋亡(P0.05);Western Blot证明甲基莲心碱可促进癌细胞中促凋亡蛋白Bax的表达,而降低抑制凋亡蛋白Bcl-2的表达(P0.05)。结论:甲基莲心碱可能通过激活线粒体凋亡相关蛋白的表达,发挥抗骨肉瘤143B的作用。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.