鼻咽癌组织CTHRC1 mRNA、c-IAP2 mRNA表达水平与临床病理特征和预后的关系分析

摘要 目的：探讨鼻咽癌组织中胶原三股螺旋重复蛋白1（CTHRC1）信使核糖核酸（mRNA）、细胞凋亡抑制蛋白2 （c-IAP2）mRNA表达与临床病理特征和预后的关系。方法：选择2016年5月至2019年1月我院收治的82例鼻咽癌患者,取活检鼻咽癌组织,采用实时荧光定量聚合酶链反应（qRT-PCR）检测CTHRC1 mRNA、c-IAP2 mRNA表达,患者治疗结束后接受定期电话随访或定期回院复查,随访持续时间为3年。比较不同临床病理特征鼻咽癌组织中CTHRC1、c-IAP2 mRNA表达差异,绘制Kaplan-Meier生存曲线,分析不同CTHRC1 mRNA、c-IAP2 mRNA表达的鼻咽癌患者3年总生存率（OS）差异。单因素和多因素Cox比例风险回归模型分析影响鼻咽癌患者预后的危险因素。结果：低度分化、TNM Ⅳ期、淋巴结转移、颅底侵犯的鼻咽癌组织中CTHRC1mRNA、c-IAP2 mRNA表达均高于高、中度分化、TNM Ⅲ期、无淋巴结转移、无颅底侵犯的鼻咽癌组织（P＜0.05）。CTHRC1mRNA高表达、c-IAP2 mRNA高表达患者3年OS低于CTHRC1mRNA低表达、c-IAP2 mRNA低表达患者（log-rank χ²=7.088、5.511,P＜0.05）。多因素Cox比例风险回归分析结果显示TNM分期IV期、淋巴结转移、高表达CTHRC1mRNA、高表达c-IAP2 mRNA是鼻咽癌患者预后不良的危险因素（P＜0.05）。结论：鼻咽癌组织 CTHRC1 mRNA、c-IAP2 mRNA表达均异常升高,高表达CTHRC1 mRNA、c-IAP2 mRNA与鼻咽癌恶性病理特征以及预后不良有关。     

类赖氨酰氧化酶2与肿瘤转移和发生

类赖氨酰氧化酶2（lysyl oxidase—like 2,LOXL2）是赖氨酰氧化酶（1ysyl oxidase,LOX）基因家族的成员之一,其表达产物能促进胶原沉积。LOXL2的过表达能促进纤维化,并与肿瘤侵袭、转移及不良预后有关。目前大部分学者认为LOXL2是一种转移促进基因,也有实验支持其是一种肿瘤抑制基因。研究发现LOXL2可以通过激活Snail／Ecadherin通路或Src／FAK通路促进转移。LOXL2有望作为肿瘤生物标志物,用于预后判断,成为一个新的治疗靶点。     

Dkk-1在肿瘤中的研究进展

DKK-1是一种分泌型糖蛋白,通过结合细胞表面受体LRP5/6、Kremen1/2在Wnt通路中起负调控作用。Dkk-1的表达受p53、MYCN、β-catenin等基因调控。Dkk-1在细胞内的异位表达能抑制多种肿瘤细胞的增殖,但有时能在促凋亡因子存在时诱发凋亡。Dkk-1在一些肿瘤中低表达,而在另一些肿瘤中高表达。Dkk-1在不同肿瘤的发生、发展以及转移这几个阶段中的表达和功能表现出复杂多重的差异。该文就Dkk-1在肿瘤中表达和功能的研究进展作一综述。     

大鼠肝癌模型中Piwil4、Mael和Ddx4基因mRNA和蛋白表达水平及其与肿瘤检测的关系

PIWI和piRNA的表达水平与肿瘤类型密切相关。Piwil2 mRNA在肝癌中的表达量比肿瘤周围的肝脏组织的表达量高。PIWI/piRNA通路基因Piwil4、Mael和Ddx4为候选癌基因,在多种癌组织中表达,而在肝癌组织中的研究尚无报道。为探讨Piwil4、Mael和Ddx4基因在肝癌组织中表达水平变化的影响,建立了大鼠肝癌模型,并提取血清用ELISA方法检测肿瘤标记物,采用实时荧光定量PCR、蛋白质免疫印迹方法和免疫组织化学方法检测正常肝组织与模型组肝组织中Piwil4、Mael和Ddx4基因mRNA转录和蛋白表达水平。结果表明,大鼠肝癌动物模型血清中肿瘤标记物含量明显升高。Piwil4、Mael和Ddx4基因mRNA及蛋白在肝癌模型组织中高表达。研究表明,肝癌模型组织中Piwil4、Mael和Ddx4基因表达水平有望作为肝癌检测的一种分子标志物。     

鱼类Ⅰ型与Ⅱ型胶原蛋白基因系统进化及其在有/无肌间骨代表鱼中的表达比较分析

研究在分析鱼类Ⅰ型与Ⅱ型胶原蛋白基因系统进化基础上, 以有肌间骨的团头鲂(Megalobrama amblycephala)和无肌间骨的尼罗罗非鱼(Oreochromis niloticus)为研究对象, 探究了Ⅰ型与Ⅱ型胶原蛋白基因在二者不同发育阶段及不同部位肌肉组织中的表达模式。系统进化分析结果显示, 胶原蛋白基因col1a1、col1a2和col2a1在有刺鱼和无刺鱼中均各自聚为一支, 团头鲂和罗非鱼3个基因的氨基酸同源性都在90%以下。不同部位肌肉组织(背部上方、尾部上方和尾部下方)的基因表达结果显示, 团头鲂col1a1a和col1a1b基因与罗非鱼该同源基因col1a1在不同肌肉组织中的表达模式存在明显差异。在团头鲂中, Ⅰ型与Ⅱ型胶原蛋白基因在背上肌肉中的表达量高于尾部; 而在罗非鱼中, 其表达模式则相反。团头鲂和罗非鱼不同发育时期的基因定量结果显示: 团头鲂col1a1a和col2a1b基因的表达在肌间骨出现以前(15 dph)和基本出现之后(50 dph)显著(P<0.01)增加, 且Ⅰ型胶原蛋白基因和col2a1b的相对表达量在不同时期差异明显, 其中col1a1a基因在50 dph的表达丰度极高; 与团头鲂相比, 罗非鱼中相应基因的表达量变化较小, 整体波动不大。研究揭示了Ⅰ型和Ⅱ型胶原蛋白在有刺鱼与无刺鱼肌肉中的表达模式, 结果表明col1a1在团头鲂和尼罗罗非鱼两种鱼类中表达模式显著不同(P<0.01), 推测其与肌间骨发育潜在相关。     

LEF-1的调控及其与肿瘤的关系研究进展

淋巴细胞增强因子（lymphoid enhancer factor,LEF-1）是Wnt信号通路中的一个重要调控因子。其异常表达对肿瘤细胞增殖和凋亡调控起关键作用。LEF-1作为Wnt信号通路核内的转录因子,还与T细胞受体α（TCRα）的增强子相互作用形成特定的构象,从而与其他因子结合共同调节基因的表达。同时,LEF-1为一多启动子基因,编码产生致瘤的全长形式的LEF-1和对Wnt信号通路起负向调控作用的截短形式的LEF-1。诸多研究结果表明肿瘤的发生发展与LEF／TCF各亚型的比例有关。就Wnt／LEF-1信号通路的调控以及与肿瘤的关系做一综述。     

LEF-1表达与肿瘤进程研究进展

淋巴增强因子-1(lymphoid enhancer factor-1,LEF-1)属于高迁移组分(HMG)家族,它与TCRα的增强子相互作用形成特定的构象,从而与其它因子结合共同调节基因的表达。LEF-1作为核内的转录因子介导Wnt信号通路,对细胞的增殖、分化和凋亡起重要作用。近年来,研究显示许多肿瘤的发生与Wnt信号通路的异常有关,而LEF-1在肿瘤的发生发展侵润过程中起重要作用。同时,LEF-1是一个多启动子基因,编码产生致瘤的全长形式的LEF-1和对Wnt信号通路起负向调控作用的截短形式的LEF-1。研究结果表明,肿瘤的发生发展与LEF/TCF各亚型的比例有关。因此,研究LEF-1的结构、功能以及其在细胞增殖、细胞存活、肿瘤的发生发展过程中的作用意义重大。该文就LEF-1的表达以及与肿瘤的关系作一综述。     

Notch3对促进胰腺星形细胞活化的基因表达及信号通路的影响

目的: 分离培养小鼠胰腺星形细胞(PSCs),检测Notch3 对促进PSCs活化的基因表达及信号通路的影响。方法: 对小鼠PSCs进行分离培养及传代。采用免疫荧光染色检测活化的小鼠PSCs中α-SMA, fibronectin及collagen I的表达;细胞分组为空白对照组(MOCK组),阴性对照组(转染Notch3 siRNA negative control,NC组),Notch3 siRNA组(转染Notch3 siRNA,N3 siRNA组)及Notch3 siRNA-1组(转染Notch3 siRNA-1,N3 siRNA-1组),提取各组总RNA,测定RNA浓度及纯度后,送至安诺优达基因科技(北京)有限公司进行转录组测序。结果: 免疫荧光结果显示,在活化的PSCs中α-SMA,fibronectin及collagen I都有明显的表达。测序结果分析表明,与NC组相比较,在N3 siRNA组与N3 siRNA-1组,α-SMA基因,collagen I基因,fibronectin基因及CTGF基因均表达下调,与胶原蛋白代谢过程相关的基因表达上调,正向调节胶原生物合成的基因表达下调,而负向调节胶原生物合成的基因表达上调,PCNA基因表达下调;在N3siRNA组与N3siRNA-1组,调节细胞聚集的基因表达下调;在细胞组分部分,细胞外基质的基因表达下调;抑制PSCs中Notch3的表达可对细胞粘附分子信号通路,MAPK信号通路及TGF-β信号通路的组成成员的基因表达产生影响。结论: 抑制Notch3的表达可抑制PSCs的活化,降低细胞增殖能力,降低迁移聚集能力及ECM合成的能力;抑制Notch3的表达可对其他的信号如细胞粘附分子信号通路,MAPK信号通路及TGF-β信号通路产生影响。     

TIEG与肿瘤的发生发展

β-转化生长因子(transforming growth factor-β,TGF-β)诱导的早期反应基因(TGF-β inducible early gene,TIEG)产物是Sp1样转录因子家族的新成员。TIEG亚家族共有3个成员:TIEG1、TIEG2和TIEG3。TIEG作为一个转录因子,能够调节多个与细胞生长分化有关的基因的表达。近年来研究发现,TIEG异常表达与多种肿瘤的发生发展有关,如乳腺癌、肺癌、胰腺癌、卵巢癌和胃癌;并且最新的研究发现,在某些抗癌药物作用过程中TIEG的表达也明显上调。这些研究结果表明,TIEG可能是一个重要的肿瘤抑制因子,因此有必要对其进行深入研究,有助于加深对肿瘤发生机制的了解。     

MicroRNA与肿瘤相关的信号转导通路

信号转导通路在细胞代谢、生长、增殖、应激、发育和凋亡等生命活动中具有极为重要的作用。干扰这些通路将可能影响细胞的正常发育, 甚至导致肿瘤。MicroRNA(miRNA)是近年来在真核生物中发现的、在转录后水平负调节基因表达的一类长度约22个核苷酸的非编码小RNA, 其靶基因数目众多, 生物学功能广泛。在多种肿瘤中发现了miRNA的异常表达, 提示后者与肿瘤发生有关, 可能机制为调控癌基因或肿瘤抑制基因的表达。此外亦发现miRNA的靶基因有许多作用于肿瘤相关的信号转导通路。miRNA在肿瘤发生过程中的重要调控功能预示其将成为人类癌症诊断和治疗方面的新星。     

CTHRC1 promotes growth,migration and invasion of trophoblasts via reciprocal Wnt/β-catenin regulation

Preeclampsia (PE) is a pregnancy complication that is characterized by high blood pressure and is associated with high maternal and fetal morbidities. At a mechanistic level, PE is characterized by reduced invasion ability of trophoblasts. Collagen triple helix repeat containing-1 (CTHRC1) is a well-known tumor-promoting factor in several malignant tumors, but its role in trophoblasts remains unknown. In this study, we characterized the expression of CTHRC1 in placenta tissue samples from PE pregnancies and from normal pregnancies. We used the trophoblasts cell lines HTR-8/SVneo and JEG-3 to investigate the role of CTHRC1 in cell migration, invasion and proliferation. Western blot, PCR and TOP/FOP luciferase activity assays were used to investigate the molecular mechanisms underlying these cell behaviors. Placenta tissue samples obtained from pregnant women with PE expressed lower levels of CTHRC1 than those of placenta tissues from women with normal pregnancies. Down-regulation of CTHRC1 impaired cell proliferation, migration and invasion of trophoblasts, while CTHRC1 overexpression promoted nuclear translocation of β-catenin, a result that was further confirmed by TOP/FOP luciferase activity assay. Our findings suggest that CTHRC1 promotes migration and invasion of trophoblasts via reciprocal Wnt/β-catenin signaling pathway. Down-regulation of CTHRC1 may be a potential mechanism underpinning the development of preeclampsia.     

miR-155-5p modulates malignant behaviors of hepatocellular carcinoma by directly targeting CTHRC1 and indirectly regulating GSK-3β-involved Wnt/β-catenin signaling

Background

Hepatocellular carcinoma (HCC) remains one of the most lethal cancers. MicroRNA-155 (miR-155) and collagen triple helix repeat containing 1 (CTHRC1) were found to be involved in hepatocarcinogenesis, but their detailed functions in HCC are unclear. Here, we aimed to investigate the underlying role of miR-155-5p and CTHRC1 in HCC.

Methods

miR-155-5p and CTHRC1 expression levels were detected by qRT-PCR, IHC and WB in HCC patients and cell lines. Dual-luciferase assay, qRT-PCR and WB were used to validate the target interaction between miR-155-5p and CTHRC1. Biological behaviors, including apoptosis, cell cycle progression, and cell proliferation, invasion and migration, were measured by flow cytometry, CCK-8 assay and Transwell tests. A xenograft model was established to examine the effects of miR-155-5p and CTHRC1 on tumor formation. WB was finally utilized to identify the role of GSK-3β-involved Wnt/β-catenin signaling in HCC growth and metastasis.

Results

Our results showed that miR-155-5p and CTHRC1 were down-regulated and up-regulated, respectively, in HCC patients and cell lines. Dual-luciferase assay verified that CTHRC1 was the direct target of miR-155-5p. Moreover, elevated miR-155-5p expression promoted apoptosis but suppressed cell cycle progression and cell proliferation, invasion and migration in vitro and facilitated tumor formation in vivo; elevated CTHRC1 expression abolished these biological effects. Additionally, miR-155-5p overexpression increased metastasis- and anti-apoptosis-related protein expression and decreased pro-apoptosis-related protein expression, while forced CTHRC1 expression conserved the expression of these proteins.

Conclusion

Altogether, our data suggested that miR-155-5p modulated the malignant behaviors of HCC by targeting CTHRC1 and regulating GSK-3β-involved Wnt/β-catenin signaling; thereby, miR-155-5p and CTHRC1 might be promising therapeutic targets for HCC patients.

     

CTHRC1 promotes osteogenic differentiation of periodontal ligament stem cells by regulating TAZ

Collagen triple helix repeat containing 1 (CTHRC1) is associated with bone metabolism. Alveolar bone has an ability to rapidly remodel itself to adapt its biomechanical environment and function. However, whether CTHRC1 is expressed in alveolar bone tissue and the role of CTHRC1 in alveolar bone remodeling remain unclear. We used orthodontic tooth movement (OTM) rat model to study the effects of CHTRC1 in alveolar bone remodeling in vivo. We found that CTHRC1 was expressed in normal physiological condition of osteocytes, bone matrix, and periodontal ligament cells in rat. During the OTM, the expression of CTHRC1, Runx2 and TAZ were increased. We further studied the effects of CTHRC1 on osteogenic differentiation of human periodontal ligament stem cells in vitro. CTHRC1 can positively regulate the expression of TAZ and osteogenic differentiation markers like Col1, ALP, Runx2 and OCN. Overexpression of CHTRC1 increased osteogenic differentiation of PDLSCs, which could be abolished by TAZ siRNA. Our results suggest that CTHRC1 plays an important role in alveolar bone remodeling and osteogenic differentiation of PDLSCs.     

PRR11 Is a Prognostic Marker and Potential Oncogene in Patients with Gastric Cancer

PRR11 is a potential candidate oncogene that has been implicated in the pathogenesis of lung cancer, however the role of PRR11 in gastric cancer is currently unclear. In the present study, we investigated the role of PRR11 in gastric cancer by evaluating its expression status in samples from a cohort of 216 patients with gastric cancer. PRR11 was found to be overexpressed in 107 (49.5%) patients by immunohistochemistry of tissue microarrays generated using the patient samples. Furthermore, PRR11 overexpression was found to correlate significantly with clinicopathologic features such as tumor invasion, tumor differentiation, and disease stage. Survival analysis of the cohort revealed that PRR11 is an independent prognostic factor for gastric cancer patients. PRR11 was stably silenced in a gastric carcinoma cell line using an shRNA-based approach, and treated cells showed decreased cellular proliferation and colony formation in vitro and cell growth in vivo, companied by decreased expression of CTHRC1 and increased expression of LXN, proteins involved in tumor progression. Evaluation of human gastric cancer samples demonstrated that PRR11 expression was also associated with increased CTHRC1 and decreased LXN expression. These data indicate that PRR11 may be widely activated in human gastric cancer and are consistent with the hypothesis that PRR11 functions as an oncogene in the development and progression of gastric cancer.     

Roles of Wnt/β-catenin signaling in the gastric cancer stem cells proliferation and salinomycin treatment

The Wnt1 protein, a secreted ligand that activates Wnt signaling pathways, contributes to the self-renewal of cancer stem cells (CSCs) and thus may be a major determinant of tumor progression and chemoresistance. In a series of gastric cancer specimens, we found strong correlations among Wnt1 expression, CD44 expression, and the grade of gastric cancer. Stable overexpression of Wnt1 increased AGS gastric cancer cells'' proliferation rate and spheroids formation, which expressed CSC surface markers Oct4 and CD44. Subcutaneous injection of nude mice with Wnt1-overexpressing AGS cells resulted in larger tumors than injection of control AGS cells. Salinomycin, an antitumor agent, significantly reduced the volume of tumor caused by Wnt1-overexpressing AGS cells in vivo. This is achieved by inhibiting the proliferation of CD44+Oct4+ CSC subpopulation, at least partly through the suppression of Wnt1 and β-catenin expression. Taken together, activation of Wnt1 signaling accelerates the proliferation of gastric CSCs, whereas salinomycin acts to inhibit gastric tumor growth by suppressing Wnt signaling in CSCs. These results suggest that Wnt signaling might have a critical role in the self-renewal of gastric CSCs, and salinomycin targeting Wnt signaling may have important clinical applications in gastric cancer therapy.     

Gli1与β-catenin相互作用促进二者在结肠癌细胞中的协同核输入

Wnt信号通路和Hedgehog（Hh）信号通路在胚胎和干细胞的发育中发挥重要作用.此外,这两条信号途径在结肠癌复发和浸润的过程也至关重要.然而,Wnt信号通路、Hedgehog信号通路二者之间具体的交互作用机制目前仍不清楚.本文发现,这两条途径的关键分子Gli1和β-联蛋白之间存在蛋白质相互作用.Gli1与β-联蛋白之间的分子相互作用有助于二者的核输入.同时发现,在肠癌细胞系中,Gli1与β-联蛋白协同上调表达. LiCl激活细胞Wnt信号通路使Gli1表达水平增加, RNA干扰抑制Wnt信号通路,Gli1的表达水平下降.同时,Gli1的过表达也提高了细胞内β-联蛋白的表达水平,并且用Hedgehog信号通路抑制剂GANT61处理细胞,降低Gli1的表达后细胞内β 联蛋白的表达相应下降.本研究揭示了Gli1 和 β-联蛋白的相互作用及二者协助核输入在Wnt、Hedgehog信号通路交互调节中发挥重要作用,Wnt、Hedgehog信号通路交互作用为大肠癌发生发展研究提供了细胞水平交互调控机制.     

Wnt/beta-catenin 信号通路相关蛋白在胃癌组织中的表达和 肿瘤转移的关系

目的：探讨了Wnt信号通路相关蛋白在胃癌组织中的表达及与肿瘤转移的关系。方法：选取2011 年6 月到2012 年6 月我 院胃癌术后47 例肿瘤标本作为研究对象,并选取同一患者的正常胃组织作为对照研究。采用实时荧光定量PCR 和Western blot 对胃癌组织和正常胃组织Wnt 信号通路相关蛋白进行分析,并分析了肿瘤转移和非转移患者Wnt信号通路相关蛋白的变化。结 果：与正常胃组织比较,胃癌组织中Wnt1、Wnt3、Wnt3a、beta-catenin、CyclinD1 和c-Myc 等分子的mRNA 水平明显上调,差异有显 著统计学意义（P<0.05）。胃癌组织中总beta-catenin 和核内beta-catenin蛋白较正常胃组织明显增加,而磷酸化beta-catenin 较正常组明显 下降、差异有显著统计学意义（P<0.05）。与非转移组比较,转移组患者胃癌组织中Wnt1、Wnt3、Wnt3a 等分子mRNA水平显著上 调,差异有统计学意义（P<0.05）。结论：Wnt信号通路异常激活在胃癌发生和癌细胞转移中发挥着重要的作用,为临床治疗提供了 一定靶点。