EphB2基因多态性与先天性肛门直肠畸形的相关性研究

目的 探讨EphB2受体基因第6外显子的-1395A/G多态性在辽宁地区汉族人群中的分布及其与先天性肛门直肠畸形(CAM)的关系.方法 采用PCR-RFLP方法,对65例CAM患儿和115名健康儿童EphB2基因-1395A/G多态位点进行基因型检测,用SHEsis在线统计软件分析等位基因频率、基因型频率及其组间差异.结果 EphB2受体基因第6外显子的-1395A/G多态A等位基因频率及AA基因型频率在CAM组(85.4%、75.4%)与正常对照组(17.0%、4.3%)间差异有显著统计学意义(P＜0.01).结论 EphB2受体基因第6外显子的-1395A/G多态与CAM存在相关性.     

EphrinB-EphB receptor signaling contributes to bone cancer pain via Toll-like receptor and proinflammatory cytokines in rat spinal cord

Treating bone cancer pain poses a major clinical challenge, and the mechanisms underlying bone cancer pain remain elusive. EphrinB-EphB receptor signaling may contribute to bone cancer pain through N-methyl-d-aspartate receptor neuronal mechanisms. Here, we report that ephrinB-EphB signaling may also act through a Toll-like receptor 4 (TLR4)-glial cell mechanism in the spinal cord. Bone cancer pain was induced by tibia bone cavity tumor cell implantation (TCI) in rats. TCI increased the expression of TLR4 and the EphB1 receptor, the activation of astrocytes and microglial cells, and increased levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). The increased expression of TLR4 and EphB1 were colocalized with each other in astrocytes and microglial cells. Spinal knockdown of TLR4 suppressed TCI-induced behavioral signs of bone cancer pain. The TCI-induced activation of astrocytes and microglial cells, as well as the increased levels of IL-1β and TNF-α, were inhibited by intrathecal administration of TLR4-targeting siRNA2 and the EphB receptor antagonist EphB2-Fc, respectively. The administration of EphB2-Fc suppressed the TCI-induced increase of TLR4 expression but siRNA2 failed to affect TCI-induced EphB1 expression. Intrathecal administration of an exogenous EphB1 receptor activator, ephrinB2-Fc, increased the expression of TLR4 and the levels of IL-1β and TNF-α, activated astrocytes and microglial cells, and induced thermal hypersensitivity. These ephrinB2-Fc-induced alterations were suppressed by spinal knockdown of TLR4. This study suggests that TLR4 may be a potential target for preventing or reversing bone cancer pain and other similar painful processes mediated by ephrinB-EphB receptor signaling.     

EphB4受体及其配体EphrinB2 mRNA在子宫内膜癌组织中的表达及意义

目的:探讨红细胞生成素诱导的肝细胞受体EphB4及其配体EphrinB2mRNA在子宫内膜癌组织中的表达及意义。方法:采用逆转录PCR(RT-PCR)法检测33例子宫内膜癌和10例正常子宫内膜组织中EphB4、EphrinB2mRNA的表达率及相对含量,并分析二者与子宫内膜癌临床病理特征的关系。结果:①EphB4mRNA在子宫内膜癌组的表达率和相对含量均高于正常子宫内膜组(P<0.05,P<0.05)。EphrinB2mRNA在子宫内膜癌组的表达率和相对含量也均高于正常子宫内膜组(P<0.05,P<0.01)。②EphB4、EphrinB2mRNA的表达与子宫内膜癌手术病理分期、组织学分级有密切关系(P<0.05)。二者表达与子宫内膜癌不同组织学类型、肌层浸润程度以及有无淋巴结转移无关(P>0.05)。结论:EphB4和EphrinB2的异常表达与子宫内膜癌发生、进展以及恶性程度有关,二者表达趋势呈现一致性,检测EphB4/EphrinB2有助于判断子宫内膜癌预后。     

EphB6受体在恶性肿瘤中的研究进展

受体酪氨酸激酶(RTKs)是一种主要的膜受体,调控细胞增殖、分化和迁移。解除RTK信号通路的管制会导致许多疾病,如癌症和发育障碍。促红细胞生成素肝细胞受体(Erythropoietin-producing hepatocellular,Eph)家族是酪氨酸激酶受体家族中最大的一个亚族,其与配体Ephrin的相互作用在生长发育和肿瘤发生过程中发挥着重要作用。研究表明,一种缺乏酪氨酸激酶活性的特殊Eph受体EphB6在乳腺癌、结直肠癌等许多恶性肿瘤中表达下降,而大量证据表明EphB6表达的缺失依赖于其启动子DNA的高甲基化,进而促进肿瘤的进展与转移。EphB6是近期研究的热点因子,本文就其目前在恶性肿瘤中研究进展做一综述。     

EphB1通过PI3K/AKT信号通路促进食管鳞状细胞癌EC-9706细胞增殖、迁移、侵袭并抑制其凋亡

目的:研究促红细胞生成素产肝细胞受体B1(erythropoietin-producing human hepatocellular receptor B1,EphB1)对食管鳞状细胞癌EC-9706细胞增殖、迁移、侵袭及凋亡的影响,并探索其可能的作用机制。方法:qRT-PCR法分析40对食管鳞状细胞癌手术患者的癌组织及邻近正常组织中EphB1表达情况,并分析其临床特征。在EC-9706细胞中分别转染si-EphB1 RNA和oe-EphB1 RNA及其阴性对照。通过qRT-PCR和蛋白免疫印迹实验检测转染效率;CCK-8实验分析EphB1对细胞活力的影响;Hoechst 33258染色和流式细胞学实验检测细胞凋亡;划痕愈合实验及Transwell侵袭实验明确细胞迁移及侵袭能力;蛋白免疫印迹实验检测EphB1蛋白、增殖相关蛋白［增殖细胞核抗原（proliferating cell nuclear antigen,PCNA）］、凋亡相关蛋白［Bad、Bcl-2、Caspase 3 及剪切型-Caspase 3（cleaved-Caspase 3）］、侵袭转移相关蛋白［基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)、基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2）、Snail、波形蛋白(Vimentin)、N-cadherin、E-cadherin］以及PI3K/AKT信号通路相关蛋白（PI3K、AKT、p-AKT）的表达水平。结果:EphB1在食管鳞状细胞癌组织及细胞系中高表达(P均＜0.05),EphB1的表达水平与食管鳞状细胞癌患者的TNM分期、有无淋巴结转移和远处转移具有显著相关性(P均＜0.05)。与对照组相比,通过细胞转染技术沉默EphB1和过表达EphB1使EC-9706细胞的增殖、迁移、侵袭能力明显降低或增强,并诱导或抑制细胞凋亡(P均＜0.05)。在蛋白水平上,si-EphB1和oe-EphB1处理EC-9706细胞48 h后,增殖相关蛋白PCNA蛋白表达水平分别显著降低或增加（P均＜0.05）,并分别增加或降低促凋亡相关蛋白Bad、cleaved-Caspase 3的表达水平（P均＜0.05）,下调或上调抗凋亡蛋白Bcl-2、Caspase 3的蛋白表达（P均＜0.05）,抑制或促进侵袭转移相关蛋白MMP-9、MMP-2、Snail、Vimentin、N-cadherin的蛋白活性,但却上调或下调 E-cadherin的表达水平（P均＜0.05）。同时,Western blotting实验结果还证实EphB1可诱导食管鳞状细胞癌EC-9706细胞中通路蛋白PI3K、p-AKT的活性增强（P均＜0.05）。结论:EphB1可能通过调控PI3K/AKT信号通路促进食管鳞状细胞癌EC-9706细胞的增殖、迁移及侵袭,并抑制其凋亡。     

左、右半结肠癌组织中EphB2的表达及其与预后相关性分析

目的:探讨人结肠癌组织中EphB2蛋白的表达情况,对比分析左、右半结肠癌组织中EphB2蛋白的表达并分析其与预后相关性.方法:选取180例结肠癌标本,应用免疫组化检测结肠癌组织及癌旁组织中EphB2蛋白的表达,并对病理参数及预后情况进行分析.结果:左半结肠癌组织中,EphB2与组织学分级、淋巴结转移及Dukes分期呈负...     

Retracted: EphB3‐targeted regulation of miR‐149 in the migration and invasion of human colonic carcinoma HCT116 and SW620 cells

microRNAs play key roles during various crucial cell processes such as proliferation, migration, invasion and apoptosis. Also, microRNAs have been shown to possess oncogenic and tumor‐suppressive functions in human cancers. Here, we describe the regulation and function of miR‐149 in colorectal cancer cell lines. miR‐149 expression patterns were detected in human colorectal cell lines and tissue samples, and then focused on its role in regulation of cell growth, migration, invasion, and its target gene identification. Furthermore, the function of the target gene of miR‐149 was analyzed in vitro and in vivo. miR‐149 expression was downregulated in human colorectal cancer HCT116 and SW620 cell lines compared to the normal colon epithelial NCM460 cell line using quantitative real‐time polymerase chain reaction methods. Further studies indicated that introduction of miR‐149 was able to suppress cell migration and invasion. Then, EphB3 was identified as a direct target gene of miR‐149 in colorectal cancer cells. Moreover, experiments in vitro showed that knockdown expression of EphB3 could suppress cell proliferation and invasion, and ectopic expression of EphB3 restored the phenotypes of CRC cell lines transfected with miR149. In addition, silencing of EphB3 significantly affected cycle progression distribution and increased apoptosis in CRC cell lines. Finally, in vivo results demonstrated that knockdown of EphB3 by siRNA inhibited tumor growth. In conclusion,the important role of miR‐149 in colorectal cancer progression suggesting that miR‐149 may serve as a therapeutic target for colorectal cancer treatment.     

结直肠癌ＥｐｈＢ６蛋白的表达及其意义

目的　探讨ＥｐｈＢ６蛋白在结直肠癌中的表达及其临床意义。方法　采用免疫组织化学染色检测８２例结直肠癌组织与癌旁组织中ＥｐｈＢ６蛋白的表达,并比较两者表达的差异;分析ＥｐｈＢ６蛋白与结直肠癌临床病理特征的关系;分析有随访资料的４９例患者ＥｐｈＢ６蛋白表达与预后的关系。结果　与癌旁组织相比,结直肠癌组织ＥｐｈＢ６蛋白表达下调４０例（４８.８％）,表达上调１４例（１７.１％）,无变化２８例（３７.１％）。结直肠癌组织ＥｐｈＢ６蛋白表达下调者多为中晚期（Ｐ＝００１５）或淋巴结转移者（Ｐ＝０.０４８）,但与肿瘤分化无明显相关。结直肠癌组织ＥｐｈＢ６蛋白表达下调组和无变化组的中位生存时间分别为３４个月和３３个月,上调组的生存时间较长（中位生存时间为未达到）,３组差异接近有统计学意义（Ｐ＝０.０５２）。结论ＥｐｈＢ６基因在结直肠癌的发展过程中可能有一定抑制肿瘤转移及发展的作用,该蛋白有可能成为结直肠癌预后的新指标。     

Coexpression of EphB4 and ephrinB2 in tumour advancement of ovarian cancers

EphB4 and ephrinB2 expressions in ovarian cancers were studied to analyse EphB4/ephrinB2 functions against clinical backgrounds. EphB4 and ephrinB2 were dominantly localised in ovarian cancer cells of all cases studied. Both the histoscores and mRNA levels of EphB4 and ephrinB2 significantly increased with clinical stages (I相似文献   

非小细胞肺癌组织中EphB4和ephrinB2的表达与CT肺灌注成像的关系

目的 探讨非小细胞肺癌(NSCLC)组织中EphB4和ephrinB2的表达及其与CT肺灌注成像的关系.方法 对31例NSCLC患者的肺内单发结节行CT灌注成像,获得血流量(BF)、血容量(BV)和最高增强值(PEI)数据.采用免疫组化SP法检测EphB4和ephrinB2在肿瘤细胞与间质血管中的表达.运用Spearman相关分析与趋势检验探讨EphB4和ephrinB2蛋白表达与NSCLC临床病理特征、灌注参数的关系.结果 31例NSCLC组织中,22例(71.0%)EphE4表达阳性,26例(83.9%)ephrinB2表达阳性,明显高于内对照(分别为24.0%和20.3%,P＜0.01).EphB4和ephrinB2的表达在肿瘤细胞中有一致性,在肿瘤间质血管中有差异性.EphB4和ephrinB2蛋白表达强度与淋巴结转移均呈低度正相关(r=0.370,P=0.045;r=0.362,P=0.047).31例NSCLC患者BF为(42.3±10.2)ml·100 mg-1·min-1,BV为(9.0±3.9)mL/100 g,PEI为(21.6±8.4)HU.相关分析显示,在NSCLC肿瘤细胞中,EphB4的表达强度分别与BF和BV呈负相关(r=-0.452,P=0.011;r=-0.408,P=0.023),与PEI无关.ephrinB2的表达强度分别与BF和BV呈正相关(r=0.516,P=0.003;r=0.448,P=0.013),与PEI无关.EphB4表达强阳性组和阳性组的BF与BV值均显著低于EphB4表达阴性组(均P＜0.01).ephrinB2表达强阳性组的BF值显著高于ephrinB2表达阳性组和阴性组(均P＜0.01),ephrinB2表达强刚性组的BV值显著高于ephrinB2表达阴性组(P＜0.01).结论 灌注成像反映功能性管腔化血管的密度差异,这种差异同时取决于管腔化血管的数量与质量.

Abstract：

Objective To investigate the role of the expression of ephrinB2 and EphB4 in non-small cell lung cancer (NSCLC), and their relationship with multi-slice spiral CT pulmonary perfusion imaging.Methods Thirty-one nodules with pathologically proven NSCLC underwent CT perfusion scan. The perfusion parameters including blood flow (BF) , blood volume (BV), peak enhancement image (PEI)were collected. The expression of ephrinB2 and EphB4 in tumor cells and interstitial vasculature were detected by immunohistochemistry. Correlation analysis and trend test were used to assess the relationship between ephrinB2/EphB4 expression and clinicopathological features, and between ephrinB2/EphB4 expression and perfusion parameters. Results Positive expression of ephrinB2 and EphB4 in the NSCLC group was 83.9%and 71.0%, respectively, significantly higher than that in the internal control group ( P ＜ 0.01 ). The expression of ephrinB2 and EphB4 was consistently in tumor parenchyma but differently in tumor vessels.The expressions of ephrinB2 and EphB4 were positively correlated with lymphatic metastasis ( P ＜ 0. 05 ).The expression of EphB4 was negatively correlated with blood flow (BF) and blood volume (BV),respectively (P ＜ 0.05 ). There was a significant positive correlation between ephrinB2 expression and BF (r =0.516,P =0. 003), and a positive correlation between ephrinB2 expression and BV (r = 0.448, P =0.013 ). The expressions of ephrinB2 and EphB4 were not correlated with PEI (P ＞0.05 ). The values of BF and BV in the high and moderate EphB4 expression groups were significantly decreased compared with that in the negative group ( P ＜ 0. 01 ). The value of BF in the high ephrinB2 expression group was significantly increased compared with that in the moderately positive group and negative group (P ＜0.01 ).The value of BV in the high ephrinB2 expression group was significantly increased compared with that in the negative group (P ＜ 0. 01 ). Conclusion The CT pulmonary perfusion imaging reflects the density difference of blood vessels with functional lumen, and such difference also depends on the quantity and quality of vasculature with functional lumen.