核黄素对人食管癌细胞株生长增殖的影响

[目的]采用细胞培养的方法研究核黄素对人食管癌细胞株生长增殖的影响,探讨核黄素营养水平与食管癌的关系。[方法]常规培养人食管癌细胞株Eca109细胞。(1)用不同浓度的核黄素作用于细胞株,MTT比色法测定Eca109细胞的增殖活性,计算细胞增殖率。(2)采用流式细胞仪法测定核黄素作用Eca109细胞后细胞周期的改变。(3)免疫组织化学法测定核黄素对Eca109细胞作用24 h Cyclin D1蛋白表达的影响。(4)HE染色以观察核黄素作用24 h对Eca109细胞的分化影响。[结果](1)不同浓度的核黄素作用于Eca109细胞后,其增殖率未见明显变化。(2)核黄素的加入使Eca109的细胞周期改变,出现G2/M期阻滞,但不促进其凋亡。(3)核黄素处理24 h后,经过免疫组化测定,食管癌细胞Cyclin D1蛋白表达无降低,与阴、阳性对照组相比差异均无统计学意义(P﹥0.05)。(4)HE染色观察各浓度组细胞形态形态无明显改变。[结论]核黄素不会影响食管癌Eca109的增殖,但可能将细胞阻滞在M期。     

凝血酶对食管癌EC109细胞增殖的影响

目的：探讨凝血酶对食管癌EC109细胞增殖的影响。方法：MTT比色法检测细胞增殖情况。凝血酶（终浓度1 U/mL）组和其作用于EC109细胞后收集的上清分别培养EC109细胞24、48、72 h。结果：凝血酶培养EC109细胞48 h后,细胞的490 nmOD值高于对照组（P〈0.05）。培养上清处理的各组与对照组比无统计学意义。结论：凝血酶有刺激EC109细胞增殖的作用。     

TGF-β superfamily co-receptors in cancer

Transforming growth factor-β (TGF-β) superfamily signaling via their cognate receptors is frequently modified by TGF-β superfamily co-receptors. Signaling through SMAD-mediated pathways may be enhanced or depressed depending on the specific co-receptor and cell context. This dynamic effect on signaling is further modified by the release of many of the co-receptors from the membrane to generate soluble forms that are often antagonistic to the membrane-bound receptors. The co-receptors discussed here include TβRIII (betaglycan), endoglin, BAMBI, CD109, SCUBE proteins, neuropilins, Cripto-1, MuSK, and RGMs. Dysregulation of these co-receptors can lead to altered TGF-β superfamily signaling that contributes to the pathophysiology of many cancers through regulation of growth, metastatic potential, and the tumor microenvironment. Here we describe the role of several TGF-β superfamily co-receptors on TGF-β superfamily signaling and the impact on cellular and physiological functions with a particular focus on cancer, including a discussion on recent pharmacological advances and potential clinical applications targeting these co-receptors.     

Impact of resveratrol on the expression of apoptosis related gene survivin and bax in human cancer cells

Objective： We explored the mechanism of apoptosis in human esophageal cancer Ecal09 cells by resveratrol. Methods： The suppressive ratio of resveratrol on Ecal09 cells proliferation was evaluated by MTT colorimetric assay and morphology was observed by transmission electron microscope. The expression of survivin and bax was analyzed by RT-PCR and Flow Cytometry （FCM）. Results： Resveratrol inhibited the growth of Ecal09 calls in a dose-and time-dependent man- ner, and the suppressive ratio arrived at 76.42%. Morphological apoptosis could be observed after treated with resveratrol.The bulk of some drug-treated cells turned small and the nuclear chromatin became condensed and rnarginated. The results determined by RT-PCR and FCM showed that resveratrol could down-regulate surviving, while up-regulate bax. Conclusion： Resveratrol could induce the apoptosis of human esophageal cancer Ecal09 cells, and its possible molecular mechanisms might be related to modulation the expression of survivin and bax.     

CD109 promotes the tumorigenic ability and metastatic motility of pancreatic ductal adenocarcinoma cells

BackgroundAccumulating evidence indicates that CD109, a glycosylphosphatidylinositol-anchored glycoprotein, is highly expressed in human epithelial carcinomas of multiple organs including the pancreas, but its functional role in carcinoma development has not yet been fully clarified. The aim of this study was to investigate the role of CD109 in the malignancy of pancreatic ductal adenocarcinoma (PDAC).MethodsPDAC specimens of 145 cases were immunostained for CD109, and correlations between CD109 expression and clinicopathological conditions were analyzed. CD109 expression in PANC-1 cells, a PDAC-derived cell line, was decreased by siRNA or shRNA and its effect on the malignancy of PANC-1 cells was examined.ResultsSuppression of CD109 expression in PANC-1 cells resulted in reduction of in vitro cell motility and tumorigenicity in xenografts. Based on these results, we investigated the relationship between CD109 expression and metastasis of PDAC using tumor tissue specimens. Among 106 recurrent cases of 145 PDAC, there was a tendency for CD109-positive cases to be accompanied by distant metastasis.ConclusionsCD109 plays a critical role in the promotion of tumorigenic ability and cellular motility relating to metastasis of PDAC cells.     

CD109, a new marker for myoepithelial cells of mammary, salivary, and lacrimal glands and prostate basal cells

The CD109 gene encodes a glycosylphosphatidylinositol (GPI)-anchored cell surface protein. Herein it is shown that CD109 is highly expressed in myoepithelial cells of mammary, salivary, and lacrimal glands; and in prostate basal cells. The anti-CD109 antibody generated by the authors was available for formalin-fixed paraffin section, and it strongly stained myoepithelial cells and basal cells but not ductal, acinar, and secretory cells in these glands. CD109 expression was negative in examined breast ductal carcinomas and prostate adenocarcinomas. These findings indicate that CD109 is a useful marker for the diagnosis of invasive breast and prostate carcinomas.     

转录因子MAZ对c1orf109基因转录表达调控的体外研究

目的:本文旨在研究转录因子Myc相关锌指蛋白(MAZ)在体外对人类1号染色体开放阅读框109(c1orf109)基因的转录表达调控。方法:体外条件下,采用凝胶电泳迁移实验筛选c1orf109基因启动子区MAZ的结合位点,并利用本室构建的由c1orf109启动子驱动的增强型绿色荧光蛋白报告载体与MAZ和转录因子特化蛋白1(Sp1)的表达质粒共转染He La细胞,24 h后,用激光共聚焦显微镜和流式细胞术检测c1orf109基因的转录表达情况。结果:c1orf109启动子区可与MAZ结合,且有与Sp1共享的结合位点;MAZ与Sp1皆可抑制c1orf109的转录表达,且Sp1的抑制作用大于MAZ(P0.05)。结论:MAZ与Sp1两个转录因子共同调控c1orf109基因在生理及病理条件的表达,且调控方向一致。这种冗余机制调控方式的存在提示该基因的精确表达调控对于细胞行使其生物学功能可能具有重要意义     

人食管癌Eca-109细胞膜蛋白分离纯化方法的研究

目的　建立人食管癌细胞株Eca 10 9细胞膜蛋白分离和初步纯化方法。方法　使用改良的Neville法提取细胞膜 ,在pH 6.3的条件下 ,用去垢剂TritonX 10 0和辛基 β 葡糖苷把细胞膜上的蛋白溶解下来 ,以超滤法纯化膜蛋白并分组。 结果　在pH6.3的条件下 ,适当的去垢剂与膜蛋白之比 (即mg去垢剂 /mg膜蛋白 ) ,使用辛基 β 葡糖苷时为 6∶1;使用TritonX 10 0时为 2∶1。超滤法将膜蛋白分为 <3KD、3～ 10KD、<10KD、10～ 3 0KD、3 0～ 10 0KD、>10 0KD共 6个组。结论　为进一步研究食管癌Eca 10 9细胞肿瘤抗原奠定基础     

莪术醇联合顺铂对人食管癌109细胞凋亡以及细胞核因子-κB 表达的影响

目的：研究中药莪术醇联合顺铂对食管癌109细胞系的增殖凋亡、核因子（NF）-κB表达的影响,探讨莪术醇抗肿瘤的分子机制。方法将不同浓度莪术醇、顺铂、莪术醇和顺铂联合作用于食管癌109细胞,用噻唑蓝比色法（MTT 法）检测细胞增殖,流式细胞仪检测细胞凋亡, Western blot 法检测作用48小时后细胞 NF-κB 蛋白的表达情况。结果不同浓度莪术醇、顺铂均对食管癌细胞均有抑制增殖、促进凋亡作用,抑制率、凋亡率呈明显浓度依耐性;联合用药后抑制率、凋亡率显著提高,差异有统计学意义（P ＜0．05）;4个浓度的莪术醇与顺铂（2．5 mg／L）作用食管癌48小时后 NF-κB 的表达量随浓度增加而下降,与对照组比较差异有统计学意义（P ＜0．05）。结论中药莪术醇对人食管癌109细胞株有明显抑制增殖,诱导凋亡的作用,其机制可能与下调NF-κB 蛋白的表达有关。     

Novel reversible selective inhibitor of nuclear export shows that CRM1 is a target in colorectal cancer cells

Colorectal cancer arises via a multistep carcinogenic process and the deregulation of multiple pathways. Thus, the simultaneous targeting of multiple pathways may be a promising therapeutic approach for colorectal treatment. CRM1 is an attractive cancer drug target, because it can regulate multiple pathways and tumor suppressor proteins. In this study, we investigated the anti-tumor activity of a novel reversible CRM1 inhibitor S109 in colorectal cancer. Our data demonstrate that S109 inhibits proliferation and induces cell cycle arrest in colorectal cancer cells. Mechanistically, we demonstrate that the activity of S109 is associated with the nuclear retention of major tumor suppress proteins. Furthermore, the Cys528 mutation of CRM1 prevented the ability of S109 to block nuclear export and inhibit the proliferation of colorectal cancer cells. Interestingly, S109 decreased the CRM1 protein level via proteasomal pathway. We argue that reversible CRM1 inhibitors but not irreversible inhibitors can induce the degradation of CRM1, because the dissociation of reversible inhibitors of CRM1 changes the conformation of CRM1. Taken together, these findings demonstrate that CRM1 is a valid target for the treatment of colorectal cancer and provide a basis for the development of S109 therapies for colorectal cancer.