SCUBE2过表达通过Wnt/β-catenin抑制结直肠癌细胞上皮-间质转化

目的:研究信号肽-CUB-EGF结构域蛋白2(SCUBE2)对结直肠癌细胞上皮-间质转化(EMT)的影响及可能的分子机制。方法:首先,用real-time PCR与Western blot法检测人结直肠癌HCT116细胞和正常人结直肠上皮FHC细胞SCUBE2的表达;HCT116细胞转染GV144-SCUBE2质粒过表达SCUBE2后,MTT、Transwell和流式细胞术分别检测其对细胞活力、迁移和凋亡的影响。转染GV144-SCUBE2质粒6 h后加入10μg/L重组转化生长因子(TGF)-β1蛋白继续培养48 h,real-time PCR或者Western blot法检测EMT标志物[E-钙黏蛋白(E-cadherin)、波形蛋白(vimentin)和Snail]、β-catenin、c-Myc和细胞周期蛋白(cyclin)D1的表达。随后,在HCT116细胞中使用Wnt/β-catenin通路激活剂氯化锂(Li Cl)或抑制剂XAV93920并加入TGF-β1且过表达SCUBE2,检测Wnt/β-catenin通路激活或阻断后对HCT116细胞EMT的影响。结果:HCT116细胞中SCUBE2的表达明显低于FHC细胞;过表达SCUBE2能够抑制HCT116细胞的活力与迁移,但是对其凋亡影响不大;SCUBE2增加了E-cadherin的表达,降低了TGF-β1诱导的vimentin、Snail、β-catenin、c-Myc和cyclin D1的表达,XAV93920增强了SCUBE2对EMT的影响,而Li Cl阻碍SCUBE2对EMT的影响。结论:过表达SCUBE2能够抑制结直肠癌细胞的生长与迁移,抑制其EMT的发生,这一过程可能是通过抑制Wnt/β-catenin信号通路发挥作用的。     

二苯乙烯苷通过PI3K/AKT通路抑制TGFβ诱导的结直肠癌上皮间质转换

目的:探讨二苯乙烯苷(2,3,5,4'-tetrahydroxy stilbene-2-Ο-β-D-glucoside,THSG)对TGFβ诱导的结直肠癌细胞上皮间质转换(epithelial-mesenchymal transition,EMT)的影响及可能的分子机制.方法:通过转化生长因子(transforming growth factorβ,TGFβ)诱导HCT116细胞发生EMT,倒置显微镜观察细胞形态变化,Western印迹检测EMT相关分子标志物的表达;划痕实验、细胞迁移实验、倒置显微镜观察不同浓度THSG干预TGFβ刺激对HCT116细胞运动、迁移能力以及形态学的影响,Western印迹检测EMT相关标志物及PI3K/AKT通路的改变.结果:TGFβ刺激HCT116细胞后,细胞由圆形变为长梭形,E-cadherin表达降低,vimentin和N-cadherin表达增加;与对照组相比,THSG可增加E-cadherin和PTEN的表达,降低vimentin和N-cadherin和p-AKT表达,同时抑制细胞的迁移及运动(F=454.723,P<0.001;F=412.161,P<0.001).结论:THSG可通过PI3K/AKT通路抑制TGFβ诱导的HCT116细胞EMT,并降低其运动迁移能力.     

同期放化疗诱导人结直肠癌细胞发生上皮-间质转化

目的:探讨放化疗抵抗的结直肠癌细胞发生上皮-间质转化(epithelial-mesenchymal transition,EMT)的意义.方法:采用5-FU化疗同期进行放疗对人结直肠癌野生型细胞(HCTll6)进行干预,诱导放化疗共同抵抗的细胞株(HCT1 16CRR)并采用克隆形成实验进行放化疗抵抗性的鉴定.高倍显微镜下观察细胞形态学变化.采用Real-time PCR和Western印迹,检测上皮表型标志物E-cadherin,间质表型标志物N-cadherin、波形蛋白(vimentin)、核转录因子(Snail)mRNA及其蛋白的表达.结果:放化疗抵抗的结直肠癌细胞发生与EMT相符的形态学改变,细胞呈纺锤体状,极性消失,并出现伪足;Real-time PCR和Western印迹结果显示E-cadherin mRNA及蛋白表达下调;N-cadherin,vimentin,Snail mRNA及蛋白表达上调,差异有统计学意义(P＜0.05).结论:放化疗抵抗后的人结直肠癌细胞发生EMT,其与结直肠癌的治疗抵抗相关.     

lncRNA RAB11B-AS1对结直肠癌SW480细胞增殖、侵袭、迁移及凋亡的影响

目的探讨lncRNA RAB11B-AS1对结直肠癌SW480细胞增殖、侵袭、迁移及凋亡的影响及可能机制。方法 qRT-PCR方法检测34例结直肠癌组织及相应的癌旁组织、人正常结肠黏膜上皮细胞NCM460与人结直肠癌细胞株SW480中lncRNA RAB11B-AS1的表达水平。构建稳定过表达RAB11B-AS1的SW480细胞系,MTT方法检测SW480细胞的增殖能力变化;流式细胞术检测SW480细胞凋亡率;Transwell检测SW480细胞侵袭能力变化,划痕实验检测SW480细胞迁移能力变化;Western blot检测SW480细胞中细胞周期蛋白1(cyclin D1)和CDK6的表达水平。结果 RAB11B-AS1在结直肠癌组织中的表达显著低于癌旁组织(P0.01);在SW480中的表达显著低于NCM460细胞(P0.01)。RAB11B-AS1过表达后,SW480细胞的增殖、侵袭与迁移能力显著降低,凋亡率显著升高,cyclin D1与CDK6的蛋白表达降低(P0.01)。结论 lncRNA RAB11B-AS1在结直肠癌组织中表达降低,过表达RAB11B-AS1能够抑制SW480细胞的增殖、迁移与侵袭,并促进凋亡。     

lncRNA PTENP1通过miR-142-5p/PTEN分子轴调控结直肠癌HCT116细胞增殖、迁移和侵袭的分子机制

目的:探讨lncRNA PTENP1(PTENP1)通过miR-142-5p/PTEN轴调控结直肠癌(CRC)HCT116细胞增殖、迁移和侵袭的分子机制.方法:采用RT-qPCR检测CRC组织和细胞系中PTENP1和miR-142-5p的表达水平;在HCT116细胞中转染pcDNA3.1-PTENP1、sh-PTENP...     

高尔基体磷酸化蛋白2对人结直肠癌细胞放射敏感性的影响

目的观察人结直肠癌细胞的放射敏感性及辐射对其高尔基磷酸化蛋白2表达的影响。方法体外培养人结直肠癌细胞株HCT116、SW480及正常结直肠细胞株FHC,检测GOLPH2基因转录及蛋白表达。选取指数生长期人结直肠癌细胞进行0、2、4、6、8、10 Gy照射,计数细胞克隆形成率,制备细胞存活曲线。应用RT-PCR和Western blotting检测2种结直肠癌细胞株在不同剂量照射时GOLPH2基因mRNA转录水平及GOLPH2蛋白表达情况。应用Transwell侵袭实验观察不同照射剂量对人结直肠癌细胞株HCT116、SW480迁移能力的影响。结果随着照射剂量在一定范围内增高时,人结直肠癌细胞株HCT116、SW480克隆形成率逐渐下降,两种细胞株中GOLPH2基因mRNA转录水平及GOLPH2蛋白表达水平均逐渐下降。不同X射线照射剂量对人结直肠癌细胞株HCT116、SW480迁移有抑制作用,并且在8Gy、10Gy时对人结肠癌细胞株迁移能力抑制最明显。结论 X射线对人结直肠癌细胞株HCT116和SW480的增殖及迁移能力均有抑制作用,可降低人结直肠癌细胞株HCT116和SW480中GOLPH2基因mRNA转录水平及GOLPH2蛋白表达水平。     

微小RNA-1273g-3p对人结直肠癌HCT116和SW480细胞增殖和迁移的影响

目的:探讨微小RNA(MicroRNA,miR)-1273g-3p对结直肠癌细胞增殖和迁移的调控作用.方法:将miR-1273g-3p模拟物分别转染人结肠癌细胞(Human colon cancer cell,HCT116)和人结肠腺癌细胞(Human colon adenocarcinoma,SW480)细胞株,通过聚合酶链反应(Polymerase chain reaction,PCR)测定miR1273g-3p的表达情况;四唑盐(Methyl thiazolyl tetrazolium,MTT)比色法检测miR-1273g-3p对细胞增殖的作用;Transwell实验检测细胞的迁移能力.结果:转染miR-1273g-3p模拟物后,结直肠癌HCT116和SW480细胞miR-1273g-3p相对表达量明显上调;MTT和Transell实验结果显示,过表达miR-1273g-3p能明显促进HCT116和SW480细胞的增殖和迁移(P<0.01).结论:miR-1273g-3p具有促进结直肠癌HCT116和SW480细胞株增殖和迁移的作用.     

CARMA3基因敲减对结肠癌细胞HCT116生长和侵袭转移的抑制

目的:探讨CARMA3基因在人结肠癌细胞HCT116生长和侵袭转移中的作用及其机制。方法:选取高表达CARMA3的人结肠癌细胞株。应用慢病毒技术敲减CARMA3基因,puromycin筛选后构建稳定转染的HCT116-sh CARMA3细胞株。Real-time PCR和Western blot鉴定mRNA和蛋白表达的抑制情况。WST-1法和RTCA S16系统分析细胞增殖情况。集落形成实验观察集落形成。流式细胞术检测细胞周期。显微镜下观察上皮-间充质转化(EMT)形态变化。划痕实验和Transwell实验检测细胞迁移与侵袭能力的改变。Western blot分析相关分子变化,探讨可能机制。结果:4株人结肠癌细胞株中HCT116细胞的CARMA3 mRNA和蛋白表达量最高,构建稳定沉默CARMA3的HCT116-sh CARMA3细胞株,其中HCT116-sh CARMA3-93细胞中CARMA3的mRNA和蛋白受抑制最明显,将其作为细胞模型。相比于对照组,HCT116-sh CARMA3-93细胞形态发生EMT逆转,其增殖、集落形成、迁移和侵袭能力明显下降(P0.01)。HCT116-sh CARMA3-93的G_0/G_1细胞所占比例明显升高,S期细胞比例相应下降(P0.05)。信号通路分子Bcl10和NF-κB表达明显下调,MALT-1变化不明显;细胞周期相关蛋白cyclin D1显著下调,cyclin A表达略有下降;侵袭转移相关分子MMP-2和MMP-9的表达下调,MMP-7未见改变,TIMP-1和TIMP-2的表达上调;EMT相关分子E-cadherin的表达水平升高,N-cadherin、Snail、Slug和Twist的表达水平呈不同程度降低。结论:CARMA3可通过改变细胞周期和侵袭转移分子的表达、调控EMT来影响结肠癌细胞HCT116的生长和侵袭转移。这可能与NF-κB信号通路发生改变有关。     

c-Met与ERKs正反馈信号环路促进结直肠癌HCT116细胞的肿瘤生成

目的探讨结直肠癌HCT116细胞肿瘤发生的分子机制。方法细胞学实验检测结直肠癌HCT116细胞中c-Met和ERKs的表达;体外结合实验检测c-Met和ERKs的相互作用;在结直肠癌HCT116细胞及裸鼠成瘤模型中,通过过表达或沉默c-Met和ERKs检测c-Met和ERKs的关系。结果细胞学实验表明,在结直肠癌HCT116细胞中c-Met及ERKs过表达。体外结合实验表明,c-Met可以和ERKs结合。细胞及动物实验表明,沉默或过表达c-Met/ERKs,可以引起ERKs/c-Met的相应变化。结论在结直肠癌HCT116细胞中,c-Met和ERKs正反馈作用促进结直肠癌的发生。     

miR-377靶向ZEB2调节结肠癌细胞迁移、侵袭和上皮-间质转化北大核心CSCD

目的:探讨miR-377对结肠癌细胞迁移、侵袭的影响及潜在作用机制。方法:RT-qPCR检测结肠癌细胞系HT29、SW480、SW620和HCT116和人小肠上皮细胞HIEC miR-377表达。将miR-377 mimic、阴性对照物、pmirGLO-wt-ZEB2、pmirGLO-mut-ZEB2转染至HCT116细胞,双荧光素酶报告实验检测miR-377与ZEB2的靶向作用,伤口愈合实验、Transwell、Western blot检测miR-377和ZEB2对细胞迁移、侵袭、上皮-间质转化相关蛋白表达的影响。结果:与HIEC细胞相比,结肠癌细胞系miR-377表达降低(P<0.05)。与对照组相比,miR-377 mimic和pmirGLO-WT-ZEB2共转染的HCT116细胞荧光素酶活性显著降低(P<0.05),但ZEB2结合位点突变逆转了miR-377 mimic对荧光素酶活性的抑制作用(P>0.05);miR-377 mimic显著降低了HCT116细胞ZEB2蛋白表达(P<0.05)。与NC+Vector组相比,miR-377+Vector组细胞划痕愈合率、侵袭细胞数显著降低(P<0.05),E-cadherin蛋白表达显著升高(P>0.05),N-cadherin、Vimentin蛋白表达显著降低(P<0.05)。与miR-377+Vector组相比,miR-377+ZEB2组细胞划痕愈合率、侵袭细胞数显著高于miR-377+Vector组(P<0.05),E-cadherin蛋白表达显著降低(P<0.05),N-cadherin、Vimentin蛋白表达显著升高(P<0.05),miR-377+ZEB2组与NC+Vector组上述指标差异无统计学意义(P>0.05)。结论:miR-377通过靶向作用于ZEB2调节结肠癌细胞上皮-间质转化相关蛋白表达,进而影响结肠癌细胞迁移、侵袭,为结肠癌临床治疗提供了新的策略。     

Schizophrenia in a North Swedish geographical isolate, 1900–1977. Epidemiology, genetics and biochemistry

Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc² (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.     

HLA in North Colombia Chimila Amerindians

HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.     

Renal dysplasia and asplenia in two sibs

A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.     

The universal muscular chamber. A basic unit of the genitourinary, cardiovascular, gastro-intestinal, skeletal, cutaneous, respiratory and other systems, endocameral hypertension; and spasm, the key to their idiopathic diseases and arthropathies

Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.     

Der Einfluß von Nahrungsmitteln und Rauchen auf die klinisch-chemische Diagnostik von Phäochromozytom,Neuroblastom und Karzinoid-Syndrom

Zusammenfassung Der Einfluß von verschiedenen Nahrungsmitteln auf Methoden zur Bestimmung von Adrenalin (AD), Noradrenalin (NA), Vanillinmandelsäure (VMS), Metanephrinen (MN), Homovanillinsäure (HVS) und 5-Hydroxyindolessigsäure (5-HIE) im 24 h-Harn zur Diagnose des Phäochromozytoms bzw. Karzinoid-Syndroms wurde untersucht. Die in die Untersuchung einbezogenen Nahrungsmittel waren: Tee, Kaffee, Mandeln, Ananas, Käse, Walnüsse, Vanillepudding, Bananen, Tomaten und Milchschokolade. Außerdem wurde der Einfluß des Zigarettenrauchens auf die Bestimmung von AD, NA, VMS und MN untersucht.Walnüsse führten zu einer starken Erhöhung der 5-HIE-Ausscheidung. Bananen erhöhten die Ausscheidung von AD, NA, VMS, MN und 5-HIE. Kaffee und Ananas bewirkten eine geringe Zunahme der MN-Werte. Rauchen von 20–30 Zigaretten/Tag beeinflußte keine der vier Variablen.Wenn die beschriebenen Methoden benutzt werden, sollte lediglich auf den Verzehr von Bananen und Walnüssen vor und während der Harnsammelperioden verzichtet werden, da die oberen Normgrenzen im Harn überschritten werden könnten. Ein Verzicht auf Kaffee und Ananas in normalen Mengen ist nicht erforderlich. Es besteht kein Anlaß, weiterhin die bisherigen umfangreichen Restriktionen der übrigen Nahrungsmittel beizubehalten.     

A comparative study of the effects of dimebon,obsidan, finoptin,and cordaron on the functional state of ischemic focus and size of necrotic zone in experimental myocardial infarction

Dimebon, an antihistamine agent, exerts a moderate antianginal effect, improving the function of ischemic focus in the myocardium and decreasing the necrotic zone in experimental myocardial infarction. Dimebon is less active than obsidan, finoptin (except for the size of the necrotic zone), and cordaron. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 122, No. 12, pp. 642–644, December, 1996     

Effects of sex steroid hormones on lipid peroxidation and glutathione antioxidant system in rat skin

Effects of estradiol and testosterone on the intensity of lipid peroxidation and contents of glutathione redox system components in the dermis and epidermis of rat skin were studied. Only estradiol induced considerable dose-dependent and tissue-specific biphasic antioxidant effects on the skin. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 12, pp. 663–666, December, 1999     

Adhesion and growth of CaCo2 cells on surface-modified PEEK substrata

A series of surface-functionalized poly(ether ether ketone) (PEEK) films has been prepared by selective wet-chemistry; they are hydroxylated polymer (PEEK-OH) obtained by reduction, aminated polymer (PEEK-[]-NH2) prepared by coupling a diisocyanate reagent to PEEKOH (PEEK-[]-NCO) followed by hydrolysis, and carboxylated and aminocarboxylated polymers (PEEK-[]-GABA and PEEK-Lysine) resulting from the coupling of aminoacids to PEEK-[]-NCO. The aminated and carboxylated substrata promoted the adhesion and growth of CaCo2 cells in the presence of serum. Fibronectin (FN), an extra-cellular matrix protein, has been covalently fixed and/or adsorbed on various PEEK substrata, in the presence or not of a polymeric surfactant (Pluronic F68). The performances of the FN-grafted substrata (PEEK-[]-FN(1) and PEEK-[]-FN(2)) were significantly higher than those of reference substrata simply coated with FN (PEEK-OH(+FN)(1) and (2), PEEK-[]-NH2(+FN)(1) and (2)), considering the adhesion and spreading of CaCo2 cells in the absence of serum. Moreover, the stability of the adherent cells on the FN-adsorbed substrata dramatically depended on the experimental conditions applied during the PEEK coating with FN.