real-time-iontophoresis-with-tetramethylammonium-to-quantify-volume

Real-time Iontophoresis with Tetramethylammonium to Quantify Volume Fraction and Tortuosity of Brain Extracellular Space

This review describes the basic concepts and protocol to perform the real-time iontophoresis (RTI) method, the gold-standard to explore and quantify the ...

New York University School of Medicine

The structural properties of brain extracellular space (ECS) are summarised by the tortuosity (lambda) and the volume fraction (alpha). To determine if these two parameters were independent, we varied the size of the ECS by changing the NaCl content to alter osmolality of bathing media for rat cortical slices. Values of lambda and alpha were extracted from diffusion measurements using the real-time ionophoretic method with tetramethylammonium (TMA+). In normal medium (305 mosmol kg(-1)), the average value of lambda was 1.69 and of alpha was 0.24. Reducing osmolality to 150 mosmol kg(-1), increased lambda to 1.86 and decreased alpha to 0.12. Increasing osmolality to 350 mosmol kg(-1), reduced lambda to about 1.67 where it remained unchanged even when osmolality increased further to 500 mosmol kg(-1). In contrast, alpha increased steadily to 0.42 as osmolality increased. Comparison with previously published experiments employing 3000 M(r) dextran to measure lambda, showed the same behaviour as for TMA+, including the same constant lambda in hypertonic media but with a steeper slope in the hypotonic solutions. These data show that lambda and alpha behave differently as the ECS geometry varies. When alpha decreases, lambda increases but when alpha increases, lambda rapidly attains a constant value. A previous model allowing cellular shape to alter during osmotic challenge can account qualitatively for the plateau behaviour of lambda.

Independence of extracellular tortuosity and volume fraction during osmotic challenge in rat neocortex.

Spreading depression (SD) and ischemia are different pathophysiological events but have similar characteristics. This study investigated whether similarity exists in the light scattering (LS) properties during SD and ischemia in rat neocortical slices. SD was induced by injection of K(+) while ischemia was simulated by removing oxygen and glucose. LS was simultaneously recorded with changes in extracellular direct current (DC) potential and extracellular space (ECS) volume. LS was measured using a photon counting fiber optic system and the ECS volume change was determined by measuring the ECS concentration of tetramethylammonium (TMA(+)). Slices maintained in normal artificial cerebrospinal fluid (ACSF) showed a consistent LS increase during SD, but exhibited two different LS behaviors during 6 min of ischemia. In eight slices, LS decreased and remained so until the end of the ischemic challenge. In another 10 slices, LS diminished initially but, after 2 min, suddenly reversed sign, accompanied by a rapid negative shift in extracellular DC potential. When 50 or 91% of Cl(-) in the ACSF was replaced by membrane-permeable propionate, LS retained its increase during SD, but always showed the sudden LS reversal during ischemia. In contrast, when Cl(-) was substituted with membrane-impermeant methylsulfate, the SD-induced LS increase was replaced by an LS decrease, and the sudden LS reversal during ischemia was absent. While the LS signal showed different characteristics during SD and ischemia, the DC potential always presented negative shifts and the ECS volume always exhibited similar decreases. These results suggest that the polarity of the LS signal is determined by the competition of at least two factors: cell swelling and anion influx.

Light scattering in rat neocortical slices differs during spreading depression and ischemia.

Glycoprotein (GP) VI, a collagen receptor, plays a important role in collagen-mediated platelet aggregation and adhesion. To date, GPVI expression has been found only in platelets and megakaryocytes. In the present studies, we have demonstrated that GPVI was also expressed in cultured human umbilical vein endothelial cells (HUVEC) at both transcript and protein levels. Using a GPVI-specific probe, a approximately 6-kb band was detected in HUVEC as well as in platelets and megakaryoblastic cell lines by Northern blotting. Using polyclonal antibodies raised against platelet GPVI peptides, the same size band (57 kDa) was labeled with convulxin (CVX) after immuo-precipitation in both HUVEC and platelet lysates. In addition, a approximately 70-kDa band was also labeled in HUVEC. Surface expression of GPVI in HUVEC was confirmed by flow cytometry with GPVI-specific IgG or by direct labeling with FITC-conjugated CVX. Since HUVEC lack FcRgamma chain that forms complex with GPVI in platelets for signaling process, the function of GPVI in vascular endothelial cells remains to be determined.

Expression of glycoprotein VI in vascular endothelial cells.

Emodin (3-methyl-1,6,8-trihydro- xyanthrax-quinone) is the main effective composition of some Chinese herbs. Previous studies showed that emodin could inhibit the proliferation of some kind of tumor cells, such as breast cancer and lung cancer, while the mechanism(s) by which emodin suppresses tumor growth remains unknown. The study was designed to investigate the inhibitory effects and mechanisms of emodin-induced cell death in human hepatoma cell HepG2.

[Apoptosis of human hepatoma HepG2 cells induced by emodin in vitro].

It has been previously shown that cilostazol (Pletal), a drug for relief of symptoms of intermittent claudication, potently inhibits cyclic nucleotide phosphodiesterase type 3 (PDE3) and moderately inhibits adenosine uptake. It elevates extracellular adenosine concentration, by inhibiting adenosine uptake, and combines with PDE3 inhibition to augment inhibition of platelet aggregation and vasodilation while attenuating positive chronotropic and inotropic effects on the heart. In the present study, we tested the hypothesis that cilostazol combined with a more potent adenosine uptake inhibitor, dipyridamole, synergistically inhibited platelet aggregation in human blood. In the presence of exogenous adenosine (1 microM), the combination of cilostazol and dipyridamole synergistically increased intra-platelet cAMP. Furthermore, cilostazol inhibited platelet aggregation in a washed platelet assay concentration-dependently with IC50s of 0.17 +/- 0.04 microM (P < 0.05 versus plus adenosine alone of 0.38 +/- 0.05 microM), 0.11 +/- 0.06 microM (P < 0.05), and 0.01 +/- 0.01 microM (P < 0.005) when combined with 1, 3, or 10 microM dipyridamole, respectively (n = 5). In whole blood, cilostazol (0.3 to 3 microM) and dipyridamole (1 or 3 microM) synergistically inhibited collagen- and ADP-induced platelet aggregation in vitro. Furthermore, the synergism was confirmed in an open-label, sequential study in healthy human subjects using ex vivo whole-blood collagen-induced platelet aggregation. Four hours after oral co-administration of cilostazol (100 mg) and dipyridamole (200 mg), platelet aggregation was inhibited by 45 +/- 17%, while no significant inhibition was observed from subjects treated with either drug alone. The combination may provide a potential treatment of arterial thrombotic disorders.

Cilostazol and dipyridamole synergistically inhibit human platelet aggregation.

Single embryonic stem cell-derived embryoid bodies for gene screening.

Lung and colon tumors were induced in A/J, C3H, and A/J X C3H (AC3) mice by administering 16 mg/kg vinyl carbamate followed by 6 weekly doses of 12 mg/kg azoxymethane (AOM). Beginning 1 week after carcinogen treatment, the mice received chemopreventive agents, dexamethasone or piroxicam, at 0.1 and 75 mg/kg in the diet, respectively. Both AOM and vinyl carbamate induces lung tumors, but only AOM induced colon tumors. The strain sensitivity for both colon and lung tumors was A/J > AC3 > C3H mice. Dexamethasone and piroxicam reduced the multiplicity of colon and lung tumors in A/J and AC3 mice, demonstrating the advantage of a combined colon and lung bioassay. The ability of budesonide, a drug that prevents mouse lung tumors, to modulate DNA methylation in vinyl carbamate-induced lung tumors was also determined. Budesonide administered for only 7 days prior to sacrifice caused a dose-dependent (0.6 to 2.4 mg/kg diet) reversal in tumors of DNA hypomethylation and hypomethylation of the insulin-like growth factor (IGF)-II gene in the differentially methylated region (DMR) 2 region of exons 4 to 5. Longer treatment with budesonide reversed hypomethylation when administered up to the time of sacrifice. These results indicate that reversal of the hypomethylation of DNA and of specific genes in lung tumors may be applicable as a surrogate end-point biomarker for chemoprevention.

Chemoprevention: mouse colon and lung tumor bioassay and modulation of DNA methylation as a biomarker.

During blood feeding, mosquitoes inject saliva containing a mixture of molecules that inactivate or inhibit various components of the hemostatic response to the bite injury as well as the inflammatory reactions produced by the bite, to facilitate the ingestion of blood. However, the molecular functions of the individual saliva components remain largely unknown. Here, we describe anopheline antiplatelet protein (AAPP) isolated from the saliva of Anopheles stephensi, a human malaria vector mosquito. AAPP exhibited a strong and specific inhibitory activity toward collagen-induced platelet aggregation. The inhibitory mechanism involves direct binding of AAPP to collagen, which blocks platelet adhesion to collagen and inhibits the subsequent increase in intracellular Ca(2+) concentration ([Ca(2+)]i). The binding of AAPP to collagen effectively blocked platelet adhesion via glycoprotein VI (GPVI) and integrin alpha(2)beta(1). Cell adhesion assay showed that AAPP inhibited the binding of GPVI to collagen type I and III without direct effect on GPVI. Moreover, intravenously administered recombinant AAPP strongly inhibited collagen-induced platelet aggregation ex vivo in rats. In summary, AAPP is a malaria vector mosquito-derived specific antagonist of receptors that mediate the adhesion of platelets to collagen. Our study may provide important insights for elucidating the effects of mosquito blood feeding against host hemostasis.

Inhibition of collagen-induced platelet aggregation by anopheline antiplatelet protein, a saliva protein from a malaria vector mosquito.

The concentration of extracellular calcium plays a critical role in synaptic transmission and neuronal excitability as well as other physiological processes. The time course and extent of local fluctuations in the concentration of this ion largely depend on its effective diffusion coefficient (D*) and it has been speculated that fixed negative charges on chondroitin sulphate proteoglycans (CSPGs) and other components of the extracellular matrix may influence calcium diffusion because it is a divalent cation. In this study we used ion-selective microelectrodes combined with pressure ejection or iontophoresis of ions from a micropipette to quantify diffusion characteristics of neocortex and hippocampus in rat brain slices. We show that D* for calcium is less than the value predicted from the behaviour of the monovalent cation tetramethylammonium (TMA), a commonly used diffusion probe, but D* for calcium increases in both brain regions after the slices are treated with chondroitinase ABC, an enzyme that predominantly cleaves chondroitin sulphate glycans. These results suggest that CSPGs do play a role in determining the local diffusion properties of calcium in brain tissue, most likely through electrostatic interactions mediating rapid equilibrium binding. In contrast, chondroitinase ABC does not affect either the TMA diffusion or the extracellular volume fraction, indicating that the enzyme does not alter the structure of the extracellular space and that the diffusion of small monovalent cations is not affected by CSPGs in the normal brain ionic milieu. Both calcium and CSPGs are known to have many distinct roles in brain physiology, including brain repair, and our study suggests they may be functionally coupled through calcium diffusion properties.

Calcium diffusion enhanced after cleavage of negatively charged components of brain extracellular matrix by chondroitinase ABC.

Adipogenesis is a complex process that involves many genes/proteins at different stages of differentiation. In order to identify genes critical for adipogenesis, we took a novel approach based on phenotype change of individual cell, to search for genes with regulatory roles in adipogenesis genome-wide in 3T3-L1 cells.

Involvement of an alternatively spliced mitochondrial oxodicarboxylate carrier in adipogenesis in 3T3-L1 cells.

&#x7EC6;&#x80DE;&#x9057;&#x4F20;&#x5B66;&#x7578;&#x53D8;&#x548C;&#x6742; (&#x9646;) &#x4E22;&#x5931;&#x7684;&#x8BB0;&#x5F55;&#x5728;&#x8BB8;&#x591A;&#x764C;&#x75C7;&#x7684; 6 &#x53F7;&#x67D3;&#x8272;&#x4F53;&#x4E0A;&#xFF0C;&#x5E76;&#x5F15;&#x5165;&#x4E00;&#x4E2A;&#x65B0;&#x6807;&#x8BB0;&#x67D3;&#x8272;&#x4F53; 6 &#x6210;&#x4E73;&#x817A;&#x764C;&#x7EC6;&#x80DE;&#x682A;&#x8C03;&#x89E3;&#x81F4;&#x7624;&#x6027;&#x7684;&#x6291;&#x5236;&#x3002;&#x5728;&#x6B64;&#x7814;&#x7A76;&#x4E2D;&#xFF0C;&#x6211;&#x4EEC;&#x63CF;&#x8FF0;&#x4E86;&#x786E;&#x5B9A;&#x7684; KIAA1949 (phostensin) &#x4F5C;&#x4E3A;&#x63A8;&#x5B9A;&#x6291;&#x764C;&#x57FA;&#x56E0;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x57FA;&#x56E0;&#x82AF;&#x7247;&#x5206;&#x6790;&#x7504;&#x522B; 25,985 &#x957F;&#x767D;&#x81F4;&#x7624;&#x6027;&#x548C;&#x8F6C;&#x79FB;&#x6027;&#x4E73;&#x817A;&#x764C;&#x7EC6;&#x80DE;&#x4E19;&#x4E8C;&#x919B;-MB-231 &#x548C; MDA/H6&#xFF0C;&#x5BFC;&#x81F4; 651 &#x5DEE;&#x5F02;&#x8868;&#x8FBE;&#x57FA;&#x56E0;&#x7684;&#x9274;&#x5B9A;&#x67D3;&#x8272;&#x4F53; 6 &#x4ECB;&#x5BFC;&#x7684;&#x6291;&#x5236;&#x3001; &#x975E;&#x81F4;&#x7624;&#x6027;&#x548C;&#x975E;&#x8F6C;&#x79FB;&#x6027;&#x884D;&#x751F;&#x7EC6;&#x80DE;&#x7EBF;&#x4E4B;&#x95F4;&#x3002;&#x4F7F;&#x7528;&#x81EA;&#x5B9A;&#x4E49;&#x7684;&#x82AF;&#x7247;&#x5305;&#x542B;&#x8FD9;&#x4E9B; 651 &#x6269;&#x589E;&#x548C; 117 &#x63A7;&#x4EF6;&#xFF0C;&#x6211;&#x4EEC;&#x786E;&#x5B9A;&#x4E86; 200 &#x7ECF;&#x5E38;&#x6C34;&#x5E73;&#x53CA;&#x57FA;&#x56E0; 10 &#x4E73;&#x817A;&#x764C;&#x7EC6;&#x80DE;&#x589E;&#x6B96;&#x53CA;&#x4F5C;&#x4E3A;&#x53C2;&#x8003;&#x4F7F;&#x7528; MDA/H6 5 &#x80BF;&#x7624;&#x7EC4;&#x7EC7;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x751F;&#x7269;&#x4FE1;&#x606F;&#x5B66;&#x5206;&#x6790;&#x900F;&#x9732;&#xFF0C;6 &#x53F7;&#x67D3;&#x8272;&#x4F53;&#x7F16;&#x7801;&#x8FD9;&#x4E9B; 200 &#x7684;&#x57FA;&#x56E0;&#xFF0C;&#x4E0E; 4 &#x4E0B;&#x8C03;&#x548C; 21 upergulation 25&#x3002;&#x5317;&#x90E8;&#x5206;&#x6790;&#x9A8C;&#x8BC1;&#x82AF;&#x7247;&#x7ED3;&#x679C;&#x548C;&#x88AB;&#x7528;&#x6765;&#x68C0;&#x6D4B;&#x7684;&#x5927;&#x5C0F;&#x548C;&#x6570;&#x91CF;&#x7684; RNA &#x7269;&#x79CD; 2 &#x7F16;&#x7801; PTK7 KIAA1949&#xFF09; &#x548C; 2 HMGN3 SFRS3&#xFF09; &#x7684;&#x4E0A;&#x8C03;&#x57FA;&#x56E0;&#x7684;&#x7EC6;&#x80DE;&#x682A;&#x3002;&#x7279;&#x522B;&#x662F;&#xFF0C;&#x5728; 6p21.33&#x3001; &#x9891;&#x7E41;&#x7684;&#x7EC6;&#x80DE;&#x9057;&#x4F20;&#x5B66;&#x5F02;&#x5E38;&#x4E0E;&#x8559;&#x5E26;&#x533A;&#x57DF;&#x7684; KIAA1949 &#x57FA;&#x56E0;&#x8FDB;&#x884C;&#x7F16;&#x7801; 4 &#x805A; (A)-RNA &#x5728;&#x591A;&#x4E2A;&#x666E;&#x901A;&#x7684;&#x7269;&#x79CD; &#xFF08;4.6&#x3001; 4&#x3001; 3 &#x548C; 1.5 kb) &#x548C;&#x4E73;&#x817A;&#x80BF;&#x7624;&#x6837;&#x672C;&#x3002;&#x57FA;&#x56E0;&#x82AF;&#x7247;&#x5206;&#x6790;&#x63ED;&#x793A;&#x4E86; KIAA1949 &#x4E0B;&#x8C03; 86.7%(n = 15&#xFF09; &#x7684;&#x4E73;&#x817A;&#x764C;&#x7EC6;&#x80DE;&#x682A;&#x548C;&#x80BF;&#x7624;&#x7EC4;&#x7EC7;&#x3002;&#x5317;&#x90E8;&#x5206;&#x6790;&#x8868;&#x660E;&#x65E0;&#x6CD5;&#x68C0;&#x6D4B;&#x548C;&#x51CF;&#x5C11;&#x8868;&#x8FBE;&#x7684; KIAA1949 100%&#xFF08;n = 10&#xFF09; &#x7684;&#x4E73;&#x817A;&#x764C;&#x7EC6;&#x80DE;&#x682A;&#x3002;&#x603B;&#x4E4B;&#xFF0C;&#x8FD9;&#x4E9B;&#x7ED3;&#x679C;&#x5F3A;&#x70C8;&#x5EFA;&#x8BAE; KIAA1949 &#x662F;&#x65B0;&#x578B;&#x5019;&#x9009;&#x4EBA;&#x4E73;&#x817A;&#x764C;&#x6291;&#x764C;&#x57FA;&#x56E0;&#x3002;

&#x65E0;&#x6CD5;&#x68C0;&#x6D4B;&#x548C;&#x51CF;&#x5C11;&#x8868;&#x8FBE;&#x7684; KIAA1949 (Phostensin) &#x4E0A;&#x5BF9;&#x4EBA;&#x7C7B;&#x4E73;&#x817A;&#x764C;&#x63ED;&#x793A;&#x7684;&#x8F6C;&#x5F55;&#x7EC4;&#x5206;&#x6790;&#x67D3;&#x8272;&#x4F53; 6p21.33 &#x7F16;&#x7801;&#x3002;

Cytogenetic aberration and loss of heterozygosity (LOH) are documented on chromosome 6 in many cancers and the introduction of a neo-tagged chromosome 6 into breast cancer cell lines mediates suppression of tumorigenicity. In this study, we described the identification of KIAA1949 (phostensin) as a putative tumor suppressor gene. Our microarray analysis screened 25,985 cDNAs between a tumorigenic and metastatic breast cancer cell line MDA-MB-231 and the chromosome 6-mediated suppressed, non-tumorigenic and non-metastatic derivative cell line MDA/H6, resulting in the identification of 651 differentially expressed genes. Using customized microarrays containing these 651 cDNAs and 117 controls, we identified 200 frequently dysregulated genes in 10 breast cancer cell lines and 5 tumor tissues using MDA/H6 as reference. Our bioinformatics analysis revealed that chromosome 6 encodes 25 of these 200 genes, with 4 downregulation and 21 upergulation. Northern analysis validated microarray results and was used to detect the size and number of RNA species of 2 downregulated (KIAA1949, PTK7) and 2 upregulated (SFRS3, HMGN3) genes in the cell lines. Particularly, the KIAA1949 gene at 6p21.33, a band region with the frequent cytogenetic aberration and LOH encodes 4 poly(A)-RNA species (4.6-, 4-, 3- and 1.5-kb) in multiple normal and breast cancer samples. Microarray analysis revealed KIAA1949 downregulation in 86.7% (n=15) of breast cancer cell lines and tumor tissues. Northern analysis demonstrated undetectable and decreased expression of KIAA1949 in 100% (n=10) of breast cancer cell lines. Taken together, these results strongly suggest KIAA1949 is a novel candidate breast cancer suppressor gene.

Undetectable and Decreased Expression of KIAA1949 (Phostensin) Encoded on Chromosome 6p21.33 in Human Breast Cancers Revealed by Transcriptome Analysis.

Aberration cytog&#xE9;n&#xE9;tique et la perte d&#x26;#39;h&#xE9;t&#xE9;rozygotie (LOH) sont document&#xE9;s sur le chromosome 6 dans de nombreux cancers et l&#x26;#39;introduction d&#x26;#39;un chromosome n&#xE9;o-&#xE9;tiquet&#xE9; 6 dans le cancer du sein des lign&#xE9;es de cellules de suppression de m&#xE9;diateur de la tumorig&#xE9;nicit&#xE9;. Dans cette &#xE9;tude, nous avons d&#xE9;crit l&#x26;#39;identification de KIAA1949 (phostensin) comme un g&#xE8;ne suppresseur de tumeur putatif. Notre analyse des micror&#xE9;seaux ADNc projet&#xE9; 25,985 entre une lign&#xE9;e cellulaire de cancer du sein tumorig&#xE8;ne et m&#xE9;tastatique MDA-MB-231 et le chromosome 6-m&#xE9;diation supprim&#xE9;, la ligne non-tumorig&#xE8;nes et non-m&#xE9;tastatique des cellules d&#xE9;riv&#xE9; MDA/H6, r&#xE9;sultant dans l&#x26;#39;identification de 651 diff&#xE9;rentiellement exprim&#xE9;s g&#xE8;nes. Utilisation de puces personnalis&#xE9;es contenant ces 651 ADNc et 117 contr&#xF4;les, nous avons identifi&#xE9; 200 g&#xE8;nes fr&#xE9;quemment surexprim&#xE9;s dans le cancer du sein de 10 lignes de cellules et tissus tumoraux en utilisant 5 MDA/H6 comme r&#xE9;f&#xE9;rence. Notre analyse bioinformatique a montr&#xE9; que le chromosome 6 code 25 de ces 200 g&#xE8;nes, r&#xE9;gulation &#xE0; la baisse avec 4 et 21 upergulation. L&#x26;#39;analyse de Northern valid&#xE9; les r&#xE9;sultats microarray et a &#xE9;t&#xE9; utilis&#xE9; pour d&#xE9;tecter la taille et le nombre d&#x26;#39;esp&#xE8;ces d&#x26;#39;ARN de 2 r&#xE9;prim&#xE9;e (KIAA1949, PTK7) et 2 r&#xE9;gul&#xE9;e &#xE0; la hausse (SFRS3, HMGN3) g&#xE8;nes dans les lign&#xE9;es cellulaires. Particulier, le g&#xE8;ne KIAA1949 &#xE0; 6p21.33, une r&#xE9;gion de bande avec l&#x26;#39;aberration cytog&#xE9;n&#xE9;tique fr&#xE9;quente et LOH code 4 poly (A) ARN-esp&#xE8;ces (4,6-, 4 -, 3 -, et 1,5 kb) dans de multiples &#xE9;chantillons de cancer du sein et normales . L&#x26;#39;analyse de puces &#xE0; ADN a r&#xE9;v&#xE9;l&#xE9; KIAA1949 r&#xE9;gulation &#xE0; la baisse dans 86,7% (n = 15) des lignes cellulaires du cancer du sein et les tissus tumoraux. L&#x26;#39;analyse a d&#xE9;montr&#xE9; une expression du Nord et une diminution des ind&#xE9;tectable KIAA1949 dans 100% (n = 10) des lignes cellulaires du cancer du sein. Pris ensemble, ces r&#xE9;sultats sugg&#xE8;rent fortement KIAA1949 est un nouveau candidat suppresseur du sein cancer du g&#xE8;ne.

Expression ind&#xE9;tectable et diminution de la KIAA1949 (Phostensin) Encoded sur le chromosome 6p21.33 dans les cancers mammaires humaines r&#xE9;v&#xE9;l&#xE9;es par l&#x26;#39;analyse du transcriptome

Zytogenetische Aberration und Verlust der Heterozygotie (LOH) auf Chromosom 6 in vielen Krebsarten dokumentiert sind und die Einf&uuml;hrung eines Neo-getaggten Chromosoms 6 in Brust-Krebs-Zelllinien vermittelt Unterdr&uuml;ckung von Tumorigenicity. In dieser Studie beschrieben wir die Identifizierung von KIAA1949 (Phostensin) als ein mutma&szlig;liches Tumor-Suppressor-Gens. Unsere Microarray-Analyse geschirmt 25.985 cDNAs zwischen einer tumorigenic und metastasierendem Brustkrebs-Krebs-Zelllinie MDA-MB-231 und das Chromosom 6-vermittelte unterdr&uuml;ckte, nicht tumorigenic und nicht-metastasierten Derivative Zelllinie MDA/H6, wodurch die Identifizierung von 651 differentiell exprimierten Genen. Mit angepassten Microarrays, die diese 651 cDNAs und 117-Steuerelemente enth&auml;lt, wir identifiziert 200 h&auml;ufig defekten Gene in 10 Brustkrebs Zell-Linien und 5 Tumor-Gewebe mit MDA/H6 als Referenz. Unsere Bioinformatik-Analyse ergab, dass das Chromosom 6 25 dieser 200 Gene, mit 4 Heraufregulation und 21 Upergulation codiert. Nordanalyse validiert Microarray-Ergebnisse und wurde verwendet, um die Gr&ouml;&szlig;e und Anzahl von RNA 2 Downregulated (KIAA1949, PTK7) und 2 herraufreguliert (SFRS3, HMGN3) Gene in den Zelllinien zu erkennen. Besonders, das KIAA1949-gen bei 6p21.33, einer Band-Region mit den h&auml;ufigen zytogenetische Aberration und LOH codiert 4 Poly (A)-RNA-Arten (4.6, 4, 3 und 1.5 kb) in mehrere Normal und Brust-Krebs-Beispiele. Microarray-Analyse ergab KIAA1949 Medikament in 86,7 % (n = 15) von Brust-Krebs-Zelllinien und Tumor-Gewebe. N&ouml;rdlichen Analyse zeigte, nicht nachweisbar und verringerten Ausdruck der KIAA1949 in 100 % (n = 10) Brust-Krebs-Zelllinien. Zusammengenommen empfehle diese Ergebnisse KIAA1949 ein Tumorsuppressor-Brustkrebsgen Roman Kandidat ist.

Nicht nachweisbar und verminderte Expression von KIAA1949 (Phostensin) auf Chromosom 6p21.33 in den menschlichen Brustkrebsen Transcriptome Analyse offenbart kodiert.

Aberrazione citogenetica e la perdita di eterozigosi (LOH) sono documentate sul cromosoma 6 in molti tipi di cancro e l&#39;introduzione di un neo-tagged cromosoma 6 in linee di cellule di cancro al seno da intermediario soppressione del tumorigenicity. In questo studio, abbiamo descritto l&#39;identificazione del KIAA1949 (phostensin) come un gene soppressore tumorale putativo. La nostra analisi microarray proiettato 25.985 cDNAs tra una linea cellulare di cancro al seno metastatico e tumorigeno MDA-MB-231 e il cromosoma 6-mediata soppressi, non inibiscono e non-metastatico derivati cellula MDA/H6, con conseguente identificazione di geni differenzialmente espressi 651. Usando i microarrays personalizzati contenenti questi 651 cDNAs e 117 controlli, abbiamo individuato 200 frequentemente geni nel cancro al seno 10 cell lines e 5 tessuti tumorali utilizzando MDA/H6 come riferimento. La nostra analisi bioinformatica ha rivelato che quel cromosoma 6 codifica 25 di questi 200 geni, con 4 downregulation e upergulation 21. Settentrionale analisi microarray risultati validati e fu usato per rilevare la dimensione e il numero di specie di RNA dei geni sovraregolati 2 (SFRS3, HMGN3) e 2 downregolata (KIAA1949, PTK7) nelle linee cellulari. In particolare, il gene KIAA1949 presso 6p21.33, una regione di banda con le frequenti aberrazioni citogenetiche e LOH codifica 4 poli (A)-specie di RNA (4,6, 4, 3 e 1,5 kb) nella pi&ugrave; normale e campioni di cancro al seno. Analisi di microarray ha rivelato KIAA1949 downregulation 86,7% (n = 15) delle linee di cellule di cancro al seno e dei tessuti tumorali. Analisi del Nord ha dimostrato non rilevabile e diminuita espressione di KIAA1949 in 100% (n = 10) di linee di cellule di cancro al seno. Presi insieme, questi risultati suggeriscono fortemente che kiaa1949 &egrave; un gene soppressore romanzo candidato al seno cancro.

Non rilevabile e diminuita espressione di KIAA1949 (Phostensin) codificata sul cromosoma 6p21.33 nei tumori al seno umano rivelato da Transcriptome analisi.

&#x7D30;&#x80DE;&#x907A;&#x4F1D;&#x5B66;&#x7684;&#x7570;&#x5E38;&#x3068;&#x30D8;&#x30C6;&#x30ED;&#x63A5;&#x5408;&#x6027;&#x306E;&#x6D88;&#x5931;&#x306F;&#xFF08;LOH&#xFF09;&#x591A;&#x304F;&#x306E;&#x764C;&#x3068;&#x816B;&#x760D;&#x539F;&#x6027;&#x306E;&#x4E73;&#x304C;&#x3093;&#x7D30;&#x80DE;&#x682A;&#x3092;&#x4EF2;&#x4ECB;&#x6291;&#x5236;&#x306B;&#x30CD;&#x30AA;&#x30BF;&#x30B0;&#x4ED8;&#x304D;6&#x67D3;&#x8272;&#x4F53;&#x306E;&#x5C0E;&#x5165;&#x306B;&#x7B2C;6&#x67D3;&#x8272;&#x4F53;&#x4E0A;&#x306B;&#x8A18;&#x8F09;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x672C;&#x7814;&#x7A76;&#x3067;&#x306F;&#x3001;&#x63A8;&#x5B9A;&#x3055;&#x308C;&#x308B;&#x816B;&#x760D;&#x6291;&#x5236;&#x907A;&#x4F1D;&#x5B50;&#x3068;&#x3057;&#x3066;KIAA1949&#xFF08;phostensin&#xFF09;&#x306E;&#x8B58;&#x5225;&#x306B;&#x3064;&#x3044;&#x3066;&#x8AAC;&#x660E;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x79C1;&#x305F;&#x3061;&#x306E;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x30A2;&#x30EC;&#x30A4;&#x89E3;&#x6790;&#x3067;&#x306F;&#x3001;&#x767A;&#x73FE;&#x5DEE;&#x306E;&#x3042;&#x308B;651&#x306E;&#x8B58;&#x5225;&#x3001;&#x305D;&#x306E;&#x7D50;&#x679C;&#x3001;&#x816B;&#x760D;&#x5F62;&#x6210;&#x304A;&#x3088;&#x3073;&#x8EE2;&#x79FB;&#x6027;&#x4E73;&#x764C;&#x7D30;&#x80DE;&#x682A;MDA-MB-231&#x304A;&#x3088;&#x3073;&#x7B2C;6&#x67D3;&#x8272;&#x4F53;&#x5A92;&#x4ECB;&#x6027;&#x306E;&#x6291;&#x5236;&#x3001;&#x975E;&#x816B;&#x760D;&#x5F62;&#x6210;&#x6027;&#x304A;&#x3088;&#x3073;&#x975E;&#x8EE2;&#x79FB;&#x6027;&#x306E;&#x6D3E;&#x751F;&#x7D30;&#x80DE;&#x682A;MDA/H6&#x9593;&#x306B;25985&#x306E;cDNA&#x3092;&#x30B9;&#x30AF;&#x30EA;&#x30FC;&#x30CB;&#x30F3;&#x30B0;&#x3057;&#x305F;&#x907A;&#x4F1D;&#x5B50;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;cDNA&#x306E;651&#x304A;&#x3088;&#x3073;117&#x306E;&#x30B3;&#x30F3;&#x30C8;&#x30ED;&#x30FC;&#x30EB;&#x3092;&#x542B;&#x3080;&#x30AB;&#x30B9;&#x30BF;&#x30DE;&#x30A4;&#x30BA;&#x3055;&#x308C;&#x305F;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x30A2;&#x30EC;&#x30A4;&#x3092;&#x7528;&#x3044;&#x3066;&#x3001;&#x30EA;&#x30D5;&#x30A1;&#x30EC;&#x30F3;&#x30B9;&#x3068;&#x3057;&#x3066;MDA/H6&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;10&#x306E;&#x4E73;&#x764C;&#x7D30;&#x80DE;&#x682A;&#x3068;5&#x816B;&#x760D;&#x7D44;&#x7E54;&#x3067;200&#x983B;&#x7E41;&#x306B;&#x8ABF;&#x7BC0;&#x4E0D;&#x5168;&#x306E;&#x907A;&#x4F1D;&#x5B50;&#x3092;&#x540C;&#x5B9A;&#x3057;&#x305F;&#x3002;&#x79C1;&#x305F;&#x3061;&#x306E;&#x30D0;&#x30A4;&#x30AA;&#x30A4;&#x30F3;&#x30D5;&#x30A9;&#x30DE;&#x30C6;&#x30A3;&#x30AF;&#x30B9;&#x89E3;&#x6790;&#x3067;&#x306F;&#x7B2C;6&#x67D3;&#x8272;&#x4F53;&#x306F;4&#x30C0;&#x30A6;&#x30F3;&#x30EC;&#x30AE;&#x30E5;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3068;21 upergulation&#x3067;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;200&#x306E;&#x907A;&#x4F1D;&#x5B50;&#x306E;25&#x3092;&#x30B3;&#x30FC;&#x30C9;&#x3057;&#x3066;&#x3044;&#x308B;&#x3053;&#x3068;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x305F;&#x3002;&#x30CE;&#x30FC;&#x30B6;&#x30F3;&#x89E3;&#x6790;&#x306F;&#x3001;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x30A2;&#x30EC;&#x30A4;&#x306E;&#x7D50;&#x679C;&#x3092;&#x691C;&#x8A3C;&#x3057;&#x3001;&#x7D30;&#x80DE;&#x682A;&#x3067;&#x30C0;&#x30A6;&#x30F3;&#x30EC;&#x30AE;&#x30E5;&#x30EC;&#x30FC;&#x30C8;2&#xFF08;KIAA1949&#x3001;PTK7&#xFF09;&#x3068;&#x4E0A;&#x65B9;&#x5236;&#x5FA1;2&#xFF08;SFRS3&#x3001;HMGN3&#xFF09;&#x907A;&#x4F1D;&#x5B50;&#x306E;RNA&#x7A2E;&#x306E;&#x30B5;&#x30A4;&#x30BA;&#x3068;&#x6570;&#x3092;&#x691C;&#x51FA;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x4F7F;&#x7528;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3057;&#x305F;&#x3002;&#x8907;&#x6570;&#x306E;&#x6B63;&#x5E38;&#x304A;&#x3088;&#x3073;&#x4E73;&#x764C;&#x30B5;&#x30F3;&#x30D7;&#x30EB;&#x4E2D;&#x306E;&#x3001;&#x7279;&#x306B;&#x3001;&#x983B;&#x7E41;&#x306B;&#x7D30;&#x80DE;&#x907A;&#x4F1D;&#x5B66;&#x7684;&#x7570;&#x5E38;&#x3068;LOH&#x306E;&#x30D0;&#x30F3;&#x30C9;&#x9818;&#x57DF;&#x306F;4&#x3064;&#x306E;&#x30DD;&#x30EA;&#xFF08;A&#xFF09;-RNA&#x7A2E;&#xFF08;&#x304A;&#x3088;&#x3073;1.5-kb&#x306E; -   - &#x3001;3 4.6&#x3001;4&#xFF09;6p21.33&#x306B;&#x304A;&#x3051;&#x308B;KIAA1949&#x907A;&#x4F1D;&#x5B50;&#x3092;&#x30B3;&#x30FC;&#x30C9;&#x3059;&#x308B;&#x3002;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x30A2;&#x30EC;&#x30A4;&#x89E3;&#x6790;&#x3067;&#x306F;&#x3001;&#x4E73;&#x764C;&#x7D30;&#x80DE;&#x682A;&#x3068;&#x816B;&#x760D;&#x7D44;&#x7E54;&#x306E;86.7&#x30D1;&#x30FC;&#x30BB;&#x30F3;&#x30C8;&#xFF08;N = 15&#xFF09;&#x306B;KIAA1949&#x30C0;&#x30A6;&#x30F3;&#x30EC;&#x30AE;&#x30E5;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x305F;&#x3002;&#x30CE;&#x30FC;&#x30B6;&#x30F3;&#x5206;&#x6790;&#x306F;&#x3001;&#x4E73;&#x764C;&#x7D30;&#x80DE;&#x682A;&#x306E;100&#xFF05;&#xFF08;N = 10&#xFF09;KIAA1949&#x306E;&#x691C;&#x51FA;&#x9650;&#x754C;&#x4EE5;&#x4E0B;&#x3068;&#x6E1B;&#x5C11;&#x3057;&#x305F;&#x767A;&#x73FE;&#x3092;&#x793A;&#x3057;&#x305F;&#x3002;&#x307E;&#x3068;&#x3081;&#x308B;&#x3068;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x7D50;&#x679C;&#x306F;&#x3001;KIAA1949&#x306F;&#x3001;&#x65B0;&#x898F;&#x5019;&#x88DC;&#x4E73;&#x764C;&#x6291;&#x5236;&#x907A;&#x4F1D;&#x5B50;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x308B;&#x3002;

KIAA1949&#x306E;&#x691C;&#x51FA;&#x3067;&#x304D;&#x306A;&#x3044;&#x3068;&#x767A;&#x73FE;&#x4F4E;&#x4E0B;&#xFF08;Phostensin&#xFF09;&#x306F;&#x3001;&#x30C8;&#x30E9;&#x30F3;&#x30B9;&#x30AF;&#x30EA;&#x30D7;&#x30C8;&#x30FC;&#x30E0;&#x89E3;&#x6790;&#x306B;&#x3088;&#x3063;&#x3066;&#x660E;&#x3089;&#x304B;&#x306B;&#x306A;&#x3063;&#x305F;&#x30D2;&#x30C8;&#x4E73;&#x764C;&#x306B;&#x304A;&#x3051;&#x308B;&#x67D3;&#x8272;&#x4F53;6p21.33&#x306B;&#x30A8;&#x30F3;&#x30B3;&#x30FC;&#x30C9;&#x3055;&#x308C;&#x305F;

Cytogenetic &#xC218; &#xCC28; &#xBC0F; heterozygosity (LOH) &#xB9CE;&#xC740; &#xC554;&#xC5D0; &#xC5FC;&#xC0C9;&#xCCB4; 6&#xC5D0; &#xBB38;&#xC11C;&#xD654; &#xD558; &#xACE0; &#xC720; &#xBC29; &#xC554; &#xC138;&#xD3EC; &#xB77C;&#xC778;&#xC5D0; &#xB124;&#xC624; &#xD0DC;&#xADF8; &#xC5FC;&#xC0C9;&#xCCB4; 6 &#xB3C4;&#xC785; &#xC911;&#xC7AC; tumorigenicity&#xC758; &#xC5B5;&#xC81C;. &#xC774; &#xC5F0;&#xAD6C;&#xC5D0;&#xC11C; &#xC6B0;&#xB9AC;&#xB294; kiaa1949&#xC758; &#xC2DD;&#xBCC4; &#xC124;&#xBA85; (phostensin) &#xC0C1; &#xC885;&#xC591; &#xC5B5;&#xC81C; &#xC720;&#xC804;&#xC790;&#xB85C;. Microarray &#xBD84;&#xC11D; tumorigenic &#xBC0F; &#xC804;&#xC774;&#xC131; &#xC720;&#xBC29;&#xC554; &#xC554; &#xC138;&#xD3EC; &#xB77C;&#xC778; MDA-MB-231 &#xC0AC;&#xC774;&#xC758; &#xC5FC;&#xC0C9;&#xCCB4; 6 &#xC911;&#xC7AC; &#xC5B5;&#xC81C;, &#xBE44; tumorigenic &#xBC0F; &#xBE44; &#xC804;&#xC774;&#xC131; &#xD30C;&#xC0DD; &#xC140; &#xB77C;&#xC778; MDA/H6, 651 &#xCC28;&#xB3D9; &#xD45C;&#xD604;&#xB41C; &#xC720;&#xC804;&#xC790;&#xC758; &#xC2DD;&#xBCC4;&#xC5D0; &#xB530;&#xB978; 25,985 cDNAs &#xC0C1;&#xC601;. 200&#xC5D0;&#xC11C; &#xC2DD;&#xBCC4;&#xC774; 651 Cdnas&#xC640; 117 &#xCEE8;&#xD2B8;&#xB864;&#xC774; &#xD3EC;&#xD568; &#xB41C; &#xC0AC;&#xC6A9;&#xC790; &#xC9C0;&#xC815;&#xB41C; microarrays&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC;, &#xC790;&#xC8FC; dysregulated &#xC720;&#xC804;&#xC790;&#xC5D0; 10 &#xC720;&#xBC29;&#xC554; &#xC138;&#xD3EC; &#xB77C;&#xC778; &#xBC0F; MDA/H6 &#xCC38;&#xC870; &#xD558; &#xC5EC; 5 &#xC885;&#xC591; &#xC870;&#xC9C1;. &#xC6B0;&#xB9AC;&#xC758; &#xC0DD;&#xBB3C; &#xC815;&#xBCF4;&#xD559; &#xBD84;&#xC11D; &#xBC1D;&#xD600; &#xADF8; &#xC5FC;&#xC0C9;&#xCCB4; 6 4 downregulation&#xC640; 21 upergulation 200&#xC774; &#xC720;&#xC804;&#xC790;&#xC758; 25&#xB97C; &#xC778;&#xCF54;&#xB529;&#xD569;&#xB2C8;&#xB2E4;. &#xBD81;&#xBD80; &#xBD84;&#xC11D; microarray &#xACB0;&#xACFC; &#xC720;&#xD6A8;&#xC131;&#xC744; &#xAC80;&#xC0AC; &#xD558; &#xACE0; &#xC140; &#xB77C;&#xC778;&#xC5D0;&#xC11C; &#xD06C;&#xAE30;&#xC640; 2 downregulated (KIAA1949, PTK7)&#xC640; 2 upregulated (SFRS3, HMGN3) &#xC720;&#xC804;&#xC790;&#xC758; RNA &#xC885;&#xB958;&#xC758; &#xC218;&#xB97C; &#xAC80;&#xC0C9; &#xD558;&#xB294; &#xB370; &#xC0AC;&#xC6A9; &#xB418;&#xC5C8;&#xB2E4;. &#xD2B9;&#xD788;, 6p21.33, cytogenetic &#xC218; &#xC0C1;&#xC6A9; &#xCC28; &#xBC0F; LOH &#xBC34;&#xB4DC; &#xC601;&#xC5ED;&#xC5D0;&#xC11C; KIAA1949 &#xC720;&#xC804;&#xC790; &#xC778;&#xCF54;&#xB529;&#xD569;&#xB2C8;&#xB2E4; 4 &#xD3F4; &#xB9AC; (A)-&#xC5EC;&#xB7EC; &#xC815;&#xC0C1;&#xC5D0;&#xC11C; RNA &#xC885; (4.6-, 4-3 &#xBC0F; 1.5 kb) &#xBC0F; &#xC720; &#xBC29; &#xC554; &#xC0D8;&#xD50C;. Microarray &#xBD84;&#xC11D; &#xACF5;&#xAC1C; 86.7%&#xC5D0;&#xC11C; KIAA1949 downregulation (n = 15) &#xC720; &#xBC29; &#xC554; &#xC138;&#xD3EC; &#xB77C;&#xC778;&#xACFC; &#xC885;&#xC591; &#xC870;&#xC9C1;. &#xBD81;&#xBD80; &#xBD84;&#xC11D; 100 %kiaa1949&#xC5D0;&#xC11C; &#xD0D0;&#xC9C0; &#xBC0F; &#xAC10;&#xC18C; &#xC2DD; &#xC2DC;&#xC5F0; (n = 10) &#xC720; &#xBC29; &#xC554; &#xC138;&#xD3EC; &#xB77C;&#xC778;. &#xC774;&#xB7EC;&#xD55C; &#xACB0;&#xACFC; &#xAC15;&#xB825; &#xD558; &#xAC8C; &#xC81C;&#xC548; KIAA1949 &#xC18C;&#xC124; &#xD6C4;&#xBCF4; &#xC720; &#xBC29; &#xC554; &#xC5B5;&#xC81C; &#xC720;&#xC804;&#xC790;&#xAC00; &#xD568;&#xAED8; &#xCC0D;&#xC740;.

Kiaa1949&#xC758; &#xD0D0;&#xC9C0; &#xBC0F; &#xAC10;&#xC18C; &#xC2DD; (Phostensin) &#xC778;&#xAC04;&#xC758; &#xC720;&#xBC29;&#xC554; Transcriptome &#xBD84;&#xC11D; &#xD558; &#xC5EC; &#xBC1D;&#xD600;&#xC5D0; &#xC788;&#xB294; &#xC5FC;&#xC0C9;&#xCCB4; 6p21.33&#xC5D0; &#xC778;&#xCF54;&#xB529;&#xB429;&#xB2C8;&#xB2E4;.

Anormalidad citogen&#xE9;tica y la p&#xE9;rdida de heterocigosidad (LOH) se documentan en el cromosoma 6, en muchos tipos de c&#xE1;ncer y la introducci&#xF3;n de un cromosoma neo-marcado 6 en el c&#xE1;ncer de mama l&#xED;neas celulares media la supresi&#xF3;n de tumorigenicidad. En este estudio, se describe la identificaci&#xF3;n de KIAA1949 (phostensin) como un gen supresor tumoral putativo. Nuestro an&#xE1;lisis de microarrays de ADNc seleccionados 25,985 entre una l&#xED;nea de c&#xE9;lulas de mama tumorig&#xE9;nico y metast&#xE1;sico MDA-MB-231 y el cromosoma 6 mediada suprimida, derivado de la l&#xED;nea de c&#xE9;lulas no tumorig&#xE9;nicas y no metast&#xE1;sico MDA/H6, dando como resultado la identificaci&#xF3;n de los 651 expresados &#x200B;&#x200B;diferencialmente los genes. El uso de microarrays personalizados que contienen estos cDNAs 651 y 117 controles, se identificaron 200 genes con frecuencia est&#xE1;n alteradas en 10 l&#xED;neas celulares de c&#xE1;ncer de mama y 5 tejidos tumorales utilizando MDA/H6 como referencia. Nuestro an&#xE1;lisis de la bioinform&#xE1;tica revel&#xF3; que el cromosoma 6 codifica a 25 de estos 200 genes, con regulaci&#xF3;n a la baja y 4 upergulation 21. El an&#xE1;lisis Northern validado resultados de microarrays y se utiliz&#xF3; para detectar el tama&#xF1;o y n&#xFA;mero de especies de ARN de downregulated 2 (KIAA1949, PTK7) y upregulated 2 (SFRS3, HMGN3) genes en las l&#xED;neas celulares. En particular, el gen KIAA1949 en 6p21.33, una regi&#xF3;n de la banda con la aberraci&#xF3;n citogen&#xE9;tica frecuente y LOH codifica 4 poli (A)-RNA especies (4,6-, 4 -, 3 - y 1,5 kb-) en m&#xFA;ltiples muestras de c&#xE1;ncer de mama y normales . El an&#xE1;lisis de microarrays revel&#xF3; KIAA1949 regulaci&#xF3;n a la baja en el 86,7% (n = 15) de las l&#xED;neas celulares de c&#xE1;ncer de mama y los tejidos tumorales. El an&#xE1;lisis Northern demostr&#xF3; expresi&#xF3;n indetectable y disminuci&#xF3;n de KIAA1949 en 100% (n = 10) de las l&#xED;neas celulares de c&#xE1;ncer de mama. En conjunto, estos resultados sugieren fuertemente KIAA1949 es una novela de mama candidato gen supresor del c&#xE1;ncer.

Expresi&#xF3;n indetectable y Disminuci&#xF3;n de la KIAA1949 (Phostensin) codificada en el cromosoma 6p21.33 en c&#xE1;nceres de mama humanos reveladas por el an&#xE1;lisis de transcriptoma

The extracellular space (ECS) consists of the narrow channels between brain cells together with their geometrical configuration and contents. Despite being only 20-60 nm in width, the ECS typically occupies 20% of the brain volume. Numerous experiments over the last 50 years have established that molecules moving through the ECS obey the laws of diffusion but with an effective diffusion coefficient reduced by a factor of about 2.6 compared to free diffusion. This review considers the origins of the diffusion barrier arising from the ECS and its properties. The paper presents a brief overview of software for implementing two point-source paradigms for measurements of localized diffusion properties: the real-time iontophoresis or pressure method for small ions and the integrative optical imaging method for macromolecules. Selected results are presented. This is followed by a discussion of the application of the MCell Monte Carlo simulation program to determining the importance of geometrical constraints, especially dead-space microdomains, and the possible role of interaction with the extracellular matrix. It is concluded that we can predict the impediment to diffusion of many molecules of practical importance and also use studies of the diffusion of selected molecular probes to reveal the barrier properties of the ECS.

Brain Extracellular Space as a Diffusion Barrier.

Telomere lengths are tightly regulated within a narrow range in normal human cells. Previous studies have extensively focused on how short telomeres are extended and have demonstrated that telomerase plays a central role in elongating short telomeres. However, much about the molecular mechanisms of regulating excessively long telomeres is unknown. In this report, we demonstrated that the telomerase enzymatic component, hTERT, plays a dual role in the regulation of telomere length. It shortens excessively long telomeres and elongates short telomeres simultaneously in one cell, maintaining the optimal telomere length at each chromosomal end for efficient protection. This novel hTERT-mediated telomere-shortening mechanism not only exists in cancer cells, but also in primary human cells. The hTERT-mediated telomere shortening requires hTERT's enzymatic activity, but the telomerase RNA component, hTR, is not involved in that process. We found that expression of hTERT increases telomeric circular DNA formation, suggesting that telomere homologous recombination is involved in the telomere-shortening process. We further demonstrated that shelterin protein TPP1 interacts with hTERT and recruits hTERT onto the telomeres, suggesting that TPP1 might be involved in regulation of telomere shortening. This study reveals a novel function of hTERT in telomere length regulation and adds a new element to the current molecular model of telomere length maintenance.

Telomerase enzymatic component hTERT shortens long telomeres in human cells.

Human telomerase reverse transcriptase (hTERT) is an essential component required for telomerase activity and telomere maintenance. Several alternatively spliced forms of hTERT mRNA have been reported in human primary and tumor cells. Currently, however, there is no sensitive and accurate method for the simultaneous quantification of multiple alternatively spliced RNA transcripts, such as in the case of hTERT. Here we show droplet digital PCR (ddPCR) provides sensitive, simultaneous digital quantification in a single reaction of two alternatively spliced single deletion hTERT transcripts (α-/β+ and α+/β-) as well as the opportunity to manually quantify non-deletion (α+/β+) and double deletion (α-/β-) transcripts. Our ddPCR method enables direct comparison among four alternatively spliced mRNAs without the need for internal standards or multiple primer pairs specific for each variant as real-time PCR (qPCR) requires, thus eliminating potential variation due to differences in PCR amplification efficiency.

Simultaneous quantification of alternatively spliced transcripts in a single droplet digital PCR reaction.

To improve the silage quality and reduce the silage additive cost, the present experiment was designed to evaluate the potential of applying the fermented juice of epiphytic lactic acid bacteria (FJLB) as an additive in alfalfa silage. The effects of FJLB on the fermentation quality, carbohydrate and protein fractions, and aerobic stability of alfalfa silage wilted under five different conditions were investigated and compared with commercial lactic acid bacteria (CLAB) and the control. The FJLB application decreased the pH value, the volatile fatty acids and non-protein nitrogen content, and the loss of sugar by 9.9%, 22.9%, 19.6% and 9.6%, respectively; it increased the lactic acid concentration by 29.5% and the aerobic stability by 17 h in comparison to the control. The FJLB application also decreased the pH value (4.44 vs. 4.66) and volatile fatty acid content (38.32 vs. 44.82) and increased the lactic acid concentration (68.99 vs. 63.29) in comparison to the CLAB-treated silage. However, the FJLB treatment had lower aerobic stability (254 h vs. 274 h) than the CLAB treatment. The FJLB application improved silage quality in comparison to the control; in addition, its effect as a fermentation stimulant may be comparable to or even better than CLAB.

Effects of different source additives and wilt conditions on the pH value, aerobic stability, and carbohydrate and protein fractions of alfalfa silage.

The objective was to investigate the capabilities of diffusional kurtosis imaging (DKI) in detection of age-related white matter (WM) changes in elderly patients with leukoaraiosis.

Assessment of severity of leukoaraiosis: a diffusional kurtosis imaging study.

Glycoprotein (GP) VI, a collagen receptor, plays an important role in collagen-mediated platelet aggregation and adhesion. To date, GPVI expression has been found only in platelets and megakaryocytes. In the present studies, we have demonstrated that GPVI was also expressed in cultured human umbilical vein endothelial cells (HUVEC) at both transcript and protein levels. Using a GPVI-specific probe, a ˜6-kb band was detected in HUVEC as well as in platelets and megakaryoblastic cell lines by Northern blotting. Using polyclonal antibodies raised against platelet GPVI peptides, the same size band (57 kDa) was labeled with convulxin (CVX) after immuno-precipitation in both HUVEC and platelet lysates. In addition, a ˜70-kDa band was also labeled in HUVEC. Surface expression of GPVI in HUVEC was confirmed by flow cytometry with GPVI-specific IgG or by direct labeling with FITC-conjugated CVX. Since HUVEC lack FcRγ chain that forms complex with GPVI in platelets for signaling process, the function of GPVI in vascular endothelial cells remains to be determined.

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Independence of extracellular tortuosity and volume fraction during osmotic challenge in rat neocortex. The Journal of physiology\| PubMed ID: 12122149	2002
Light scattering in rat neocortical slices differs during spreading depression and ischemia. Brain research\| PubMed ID: 12376191	2002
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Inhibition of collagen-induced platelet aggregation by anopheline antiplatelet protein, a saliva protein from a malaria vector mosquito. Blood\| PubMed ID: 18056842	2008
Calcium diffusion enhanced after cleavage of negatively charged components of brain extracellular matrix by chondroitinase ABC. The Journal of physiology\| PubMed ID: 19546165	2009
Involvement of an alternatively spliced mitochondrial oxodicarboxylate carrier in adipogenesis in 3T3-L1 cells. Journal of biomedical science\| PubMed ID: 19825180	2009
Undetectable and Decreased Expression of KIAA1949 (Phostensin) Encoded on Chromosome 6p21.33 in Human Breast Cancers Revealed by Transcriptome Analysis. Journal of Cancer\| PubMed ID: 20842223	2010
Brain Extracellular Space as a Diffusion Barrier. Computing And Visualization In Science\| PubMed ID: 23172993	2011
Telomerase enzymatic component hTERT shortens long telomeres in human cells. Cell cycle (Georgetown, Tex.)\| PubMed ID: 24721976	2014
Simultaneous quantification of alternatively spliced transcripts in a single droplet digital PCR reaction. BioTechniques\| PubMed ID: 24924392	2014
Effects of different source additives and wilt conditions on the pH value, aerobic stability, and carbohydrate and protein fractions of alfalfa silage. Animal science journal = Nihon chikusan Gakkaiho\| PubMed ID: 27147561	2017
Assessment of severity of leukoaraiosis: a diffusional kurtosis imaging study. Clinical imaging\| PubMed ID: 27317218	2016
Expression of glycoprotein VI in vascular endothelial cells. Platelets\| PubMed ID: 28447516	2003

Lian Tao

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