Reduced expression ofβ-catenin inhibitor Chibby in colon carcinoma cell lines

AIM: To analyse the Chibby expression and its function in colon carcinoma cell lines and colorectal carcinoma (CRC). METHODS: Chibby expression levels were investigated by quantitative RT-PCR in a panel of seven different colon carcinoma cell lines. By sequencing, we analysed mutational status of Chibby. To test whether Chibby exhibited effects onβ-catenin signalling in colon carcinoma cells, we transfected SW480 cells with Chibby expression plasmid and, subsequently, analysed activity of p-catenin and tested for alterations in cellular phenotype. In addition, we examined Chibby mRNA levels in samples of colorectal carcinomas and adjacent normal tissues by using quantitative RT-PCR and hybridised gene chips with samples from CRC and normal tissues. RESULTS: Chibby mRNA expression was strongly down-regulated in colon carcinoma cell lines in comparison to normal colon epithelial cells and no mutation in any of the examined colon carcinoma cell lines was found. Further, we could show that Chibby inhibited p-catenin activity in TOPflash assays when over-expressed in SW480 cells. Proliferation and invasion assays with Chibby transfected SW480 cells did not reveal profound differences compared to control cells. In contrast to these in vitro data, quantitative RT-PCR analyses of Chibby mRNA levels in CRC tumor samples did not show significant differences to specimens in adjacent non-cancerous tissue. Consistent with these findings, gene chips analysing tissue samples of tumors and corresponding normal tissue did not show altered Chibby expression CONCLUSION: Altered Chibby expression might be observed in vitro in different colon carcinoma cell lines. However, this finding could not be confirmed in vitro in CRC tumors, indicating that Chibby is not likely to promote CRC tumor development or progression. As Chibby is an important inhibitor ofβ-catenin signalling, our data implicate that the usability of colon carcinoma cell lines for in vitro studies analysing the Wnt/β-catenin pathway in colorectal carcinoma needs extensive verification.

Migration-associated secretion of melanoma inhibitory activity at the cell rear is supported by KCa3.1 potassium channels

恶意的黑瘤，由侵略本地生长和转移的早形成描绘了，是皮癌症的最好攻击的类型。黑瘤禁止的活动(MIA ) ，由恶意的黑瘤房间藏匿了，与房间粘附受体， integrins 伪 4 尾 1 和伪 5 尾 1，便于的房间分开和转移的支持的形成交往。在现在的学习，我们证明那 MIA 分泌物被限制到移植房间的后面的目的，当在非移居的房间 MIA 在肌动朊外皮积累时。MIA 蛋白质包括上衣蛋白质建筑群拿一条常规能分泌的小径我(COPI )- 并且上衣蛋白质建筑群 II (COPII ) 依赖者蛋白质运输到房间圆周，在它的最后的版本取决于细胞内部的 Ca2+ 离子的地方。有趣地，激活 Ca2+ 的 K+ 隧道，亚科 N，成员 4 (KCa3.1 ) ，知道在移植房间的后面的结束活跃，被发现支持 MIA 分泌物。分泌物被特定的 KCa3.1 隧道禁止者 TRAM-34 并且由隧道的主导否定的异种的表示减少。在摘要，我们阐明了到房间尾部并且也的 MIA 蛋白质的联系迁居的运输揭示了 KCa3.1 钾隧道由支持房间移植的新机制。

Role of soluble factors and three-dimensional culture in in vitro differentiation of intestinal macrophages

AIM: To examine the factor(s) involved in differentiation of intestinal macrophages (IMACs) using a recently established in vitro model. METHODS: To test whether soluble or membrane bound factors induce IMAC-differentiation, freshly elutriated monocytes (MO) were incubated with conditioned media or cell membranes of intestinal epithelial cells (IEC) or cultured with IEC in transwell systems. To determine the importance of an active migration of MO, three- dimensional aggregates from a 1:1-mixture of MO and IEC were examined by immunohistochemistry and flow cytometry. Apoptosis was examined by caspase-3 Western blots. Extracellular matrix production in differentiation models was compared by immunohistochemistry. RESULTS: IMAC differentiation was observed in a complex three-dimensional co-culture model (multicellular spheroid, MCS) with IEC after migration of MO into the spheroids. By co-culture of MO with conditioned media or membrane preparations of IEC no IMAC differentiation was induced. Co-culture of MO with IEC in transwell- cultures, with the two cell populations separated by a membrane also did not result in intestinal-like differentiation of MO. In contrast to IEC-spheroids with immigrating MO in mixed MCS of IEC and MO only a small subpopulation of MO was able to survive the seven day culture period. CONCLUSION: Intestinal-like differentiation of MO in vitro is only induced in the complex three-dimensional MCS model after immigration of MO indicating a roleof cell-matrix and/or cell-cell interactions during the differentiation of IMACs.

Serological pattern “anti-HBc alone”:Characterization of 552 individuals and clinical significance

瞄准：调查流行和临床的意义“ anti-HBc 独自一个”在病人的一张 unselected 人口和在南部的德国的一所大学医院的雇员。方法：有模式的所有个人“ anti-HBc 独自一个”在 82 瞬间的时间跨度上被登记。HBV-DNA 在浆液和肝样品被测量，并且临床的图表被考察。结果：五百和 52 个个人是“ anti-HBc 独自一个”(3004 anti-HBc 积极个人；18.4%) ，并且这个模式比女性更经常影响了男性(20.5%)(15.3% ；P【0.001 ) 。HBV-DNA 在 545 中的 44 个的浆液被检测“ anti-HBc 独自一个”个人(8.1%) ，并且在石蜡，在 39 个病人中的 16 个的嵌入的肝织物测试了(41.0%) 。在 HBV 染色体的察觉和肝损坏的生物化学、超声或组织学的符号的存在之间没有协会。38 “ anti-HBc 独自一个”有肝硬化或主要的肝癌的病人有至少一个另外的风险因素。HCV-coinfection 在 20.4% 所有个人与是在场的“ anti-HBc 独自一个”并且唯一的因素与更坏的临床的结果被联系。结论：在一个 HBV 低流行区域，没有证据被发现那 HBV 独自一个在个人引起严重的肝损坏与“ anti-HBc 独自一个”。为这些个人的管理的建议被给。

Identification of novel sense and antisense transcription at the TRPM2 locus in cancer

在癌症，在染色体的体积成为 hypomethylated 的地方，可能导致能影响关键 oncosuppressor 基因的功能的 antisense 抄本的产生的 transcriptional 噪音有增加，这被建议了，最终导致恶意的转变。这里，我们描述一个充实黑瘤的 antisense 抄本的计算鉴定， TRPM2 -- 同样，在 TRPM2 的地点以内印射，能够调停的一条离子隧道危险性到房间死亡。TRPM2 的分析 -- 当 genomic 区域在另一个充实肿瘤的 TRPM2 抄本的一样的区域显示了存在， TRPM2-TE，定位了与 TRPM2 分享的一个 CpG 岛到对面 -- 作为。量的 PCR 实验证实了那 TRPM2 -- 作为并且 TRPM2-TE 抄本在黑瘤是起来调整的，并且他们的激活与分享的 CpG 岛的 methylation 地位一致。TRPM2-TE 的功能的大美人，以及野类型的 TRPM2 的在表示上，增加的黑瘤危险性到 apoptosis 和坏死。最后，在另外的癌症类型的表示分析显示了那 TRPM2 -- 作为并且 TRPM2-TE 在表示上可能比期望有一个甚至更宽的角色，在识别新潜力增强我们的计算途径的关联治疗学的目标。

Ephrin-B2 is differentially expressed in the intestinal epithelium in Crohn's disease and contributes to accelerated epithelial wound healing in vitro

AIM: Eph receptor tyrosine kinases and their membrane bound receptor-like ligands, the ephrins, represent a bi-directional cell-cell contact signaling system that directs epithelial movements in development. The meaning of this system in the adult human gut is unknown. We investigated the Eph/ephrin mRNA expression in the intestinal epithelium of healthy controls and patients with inflammatory bowel disease (IBD).METHODS: mRNA expression profiles of all Eph/ephrin family members in normal small intestine and colon were established by real-time RT-PCR. In addition, differential expression in IBD was investigated by cDNA array technology, and validated by both real-time RT-PCR and immunohistochemistry. Potential effects of enhanced EphB/ephrin-B signaling were analyzed in an in vitro IEC-6 cell scratch wound model.RESULTS: Human adult intestinal mucosa exhibits a complex pattern of Eph receptors and ephrins. Beside the known prominent co-expression of EphA2 and ephrinA1,we found abundantly co-expressed EphB2 and ephrin-B1/2.Interestingly, cDNA array data, validated by real-time PCR and immunohistochemistry, showed upregulation of ephrin-B2 in both perilesional and lesional intestinal epithelial cells of IBD patients, suggesting a role in epithelial homeostasis. Stimulation of ephrin-B signaling in ephrinB1/2 expressing rat IEC-6-cells with recombinant EphB1Fc resulted in a significant dose-dependent acceleration of wound closure. Furthermore, fluorescence microscopy showed that EphB1-Fc induced coordinated migration of wound edge cells is associated with enhanced formation of lamellipodial protrusions into the wound, increased actin stress fiber assembly and production of laminin at the wound edge.CONCLUSION: EphB/ephrin-B signaling might represent a novel protective mechanism that promotes intestinal epithelial wound healing, with potential impact on epithelial restitution in IBD.

A new mission for an old anti-cancer drug:harnessing hepatocyte-specific metabolic pathways against liver tumors

In a recent study published in Nature Cancer,Shi et al.reported the identification of the small molecule YC-1 as the selective drug against primary liver tumor cells due to the specific expression of sulfotransferase family 1A member 1(SULT1A1)in hepatocytelineage cells.1 This study offered new insights into repurposing an old anti-cancer drug via harnessing hepatocyte-specific metabolic enzymes to treat primary liver tumors.