Induction of epithelial-mesenchymal transition (EMT) in human hepatocellular carcinoma after radiotherapy

Objective: Epithelial-mesenchymal transition (EMT) is a critical early event for the invasion and metastasis of many carcinomas. In the present study, we examined EMT markers in the residual cancer cells of hepatocellular carcinoma (HCC) after radiotherapy. Methods: Eight patients with large HCC who underwent hepatectomy with preoperative radiothera- py were studied. The expressions of E-cadherin and vimentin were determined immunohistochemically in the residual cancer cells of HCC following radiotherapy, and also in the pre-radiotherapy biopsy cancer cells. Results: Histological analysis showed that some residual cancer cells of HCC displayed an elongated spindle or fibroblast-like shape. The expression of E- cadherin was markedly reduced or negative in the spindle residual cancer cells, but the expression of vimentin significantly in- duced. However, the above changes were not found in the pre-radiotherapy biopsy cancer cells. Conclusion: EMT is induced in the residual cancer cells of HCC following radiotherapy, which may facilitate the systemic dissemination of cancer cells.

Re-evaluating the role of epithelial-mesenchymal-transition in cancer progression

Epithelial-mesenchymal transition（EMT） and mesenchymal-epithelial transition（MET） are essential for embryonic development and also important in cancer progression. In a conventional model, epithelial-like cancer cells transit to mesenchymal-like tumor cells with great motility via EMT transcription factors; these mesenchymallike cells migrate through the circulation system, relocate to a suitable site and then convert back to an epithelial-like phenotype to regenerate the tumor. However, recent findings challenge this conventional model and support the existence of a stable hybrid epithelial/mesenchymal（E/M） tumor population. Hybrid E/M tumor cells exhibit both epithelial and mesenchymal properties, possess great metastatic and tumorigenic capacity and are associated with poorer patient prognosis. The hybrid E/M model and associated regulatory networks represent a conceptual change regarding tumor metastasis and organ colonization. It may lead to the development of novel treatment strategies to ultimately stop cancer progression and improve disease-free survival.

Transcriptional Factor Snail Mediates Epithelial-Mesenchymal Transition in Human Bronchial Epithelial Cells Induced by Silica

Epithelial-mesenchymal transition （EMT） plays an important role in fibrotic diseases. We have previously showed that silica induces EMT in human bronchial epithelial cells （BECs）; however, the underlying mechanism of silica-induced EMT is poorly understood. In the present study, we investigated the role of Snail in silica-induced EMT in human BECs in vitro. Human BECs were treated with silica at various concentrations and incubation times. Then MTr assay, western blot, electrophoretic mobility shift assay （EMSA）, and small interfering RNA （siRNA） transfection were performed. We found that silica increased the expression and DNA binding activity of Snail in human BECs. SNAI silica-induced expression siRNA upregulated the siRNA inhibited the of Snail. Moreover, SNAI expression of epithelial marker E-cadherin, but attenuated the expression of mesenchymal marker a-smooth muscle actin and vimentin in silica-stimulated cells. These results suggest that Snail mediates the silica-induced EMT in human BECs.

Strategies for Synchronous and Multiple Metastatic Liver Tumors Designed from Epithelial-Mesenchymal Transition Concept

At some point in the natural course of colorectal cancer up to 50% of patients will develop metastasis to the liver and it is one of the most critical effects for patient prognosis. The incidence of synchronous liver metastasis has been detected at around 20% - 25%, but the optimal timing of surgical resection remains controversial. Neoadjuvant chemotherapy has also been found to be beneficial not only for initially unresectable but also resectable synchronous metastases. Then, traditional surgical strategies of hepatic resection in accordance with past chemotherapeutic regimens have been used decreasingly over the past several years. This review will primarily discuss treatments in association with the recent developed chemotherapeutic regimens and surgical procedure from the clinical data and the concept for epithetlial-mesenchymal transition, which has recently been studied to elucidate mechanisms of the liver metastatic process.

Effect of Rapamycin on TGF-β_1-induced epithelial-mesenchymal transition in LoVo colonic adenocarcinoma cells

Objective： To investigate the effect of Rapamycin on epithelial-mesenchymal transition（EMT） of LoVo colonic adenocarcinoma cells in vitro. Methods：Cultured LoVo colonic adenocarcinoma cells were divided into three groups： negative control group, EMT-inducing group（TGF-β1） and EMT-interfering group（TGF-β1 plus Rapamycin）. E-cadherin expression in LoVo cells was detected by Western Blot, while the expression of vimentin was evaluated through immunocytochemistry. The Snail mRNA in LoVo cells was examined by RT- PCR. Results：TGF-β1 induced LoVo cell switching from polygonal to spindle-shaped. TGF-β1 enhanced the expression of vimentin, but lowered the level of E-cadhefin. In contrast, Rapamycin impaired the transition induced by TGF-β1. Rapamycin dramatically abrogated TGF-β1-induced vimentin expression and restored E-cadherin expression in LoVo cells. Rapamycin significantly repressed the upregulation of Snail mRNA expression induced by TGF-β1. Conclusion：Rapamycin dramatically abrogated TGF-β1 induced Snail mRNA expression in LoVo cells, hence inhibiting EMT of these cells in vitro.

The roles of micro RNAs and epithelial-mesenchymal transition in colorectal cancer metastasis

Colorectal cancer(CRC) is the second most common cause of cancer death worldwide. Distant metastasis is the major cause of death in patients with CRC. During progression to metastasis in which malignant cells disseminate from the primary tumor to seeding other organs, a multistep process is involved. Cancer cells proliferate, invade microenvironment, enter into the blood circulation, then survive and colonize into distant organs. Micro RNAs(mi RNAs) and epithelial-mesenchymal transition(EMT) are key regulators and mechanism in tumorigenesis and cancer metastasis. We review the roles of EMT and micro RNAs, especially EMT related micro RNAs in the metastatic pathway of CRC. Micro RNAs provide us a set of potential therapeutic applications and molecular target for CRC.

MiR-663a Inhibits Radiation-Induced Epithelium-to-Mesenchymal Transition by Targeting TGF-β1

Objective miR-663 a has been reported to be downregulated by X-ray irradiation and participates in radiation-induced bystander effect via TGF-β1.The goal of this study was to explore the role of mi R-663 a during radiation-induced Epithelium-to-mesenchymal transition(EMT).Methods TGF-β1 or IR was used to induce EMT.After mi R-663 a transfection,cell migration and cell morphological changes were detected and the expression levels of mi R-663 a,TGF-β1,and EMT-related factors were quantified.Results Enhancement of cell migration and promotion of mesenchymal changes induced by either TGF-β1 or radiation were suppressed by mi R-663 a.Furthermore,both X-ray and carbon ion irradiation resulted in the upregulation of TGF-β1 and downregulation of mi R-663 a,while the silencing of TGF-β1 by mi R-663 a reversed the EMT process after radiation.Conclusion Our findings demonstrate an EMT-suppressing effect by mi R-663 a via TGF-β1 in radiationinduced EMT.

Silencing Neuropilin 1 gene reverses TGF-β1-induced epithelial mesenchymal transition in HGC-27 gastric cancer cell line

Objective The aim of this study was to determine Neuropilin 1(NRP1)contribution to transforming growth factorβ1(TGF-β1)-induced epithelial mesenchymal transition(EMT)of HGC-27 gastric cancer cells and study its mechanism.Methods In this study,TGF-β1 was used to induce EMT in HGC-27 cells.Further,these cells were stably transfected with siRNA targeting NRP1.Wound healing and transwell assays were used to measure cell migration and invasion,respectively.NRP1 and EMT markers were measured using quantitative real time reverse transcription polymerase chain reaction and western blotting.Results Exposure of TGF-β1 conferred a fibroblastic-like shape to cancer cells and significantly increased the expression of NRP1 in HGC-27 cells.TGF-β1 subsequently promoted migration and invasion of HGC-27 cells.Furthermore,silencing NRP1 inhibited the invasion and migration of TGF-β1-induced cells undergoing EMT.Conclusion Silencing NRP1 can inhibit cell migration,invasion,and metastasis and reverse the TGF-β1-induced EMT process of gastric cancer.

Beta-elemene inhibits the growth of KDM6A-null bladder cancer cells by suppressing the epithelial-mesenchymal transition

Background:Bladder cancer is a highly prevalent and lethal malignant tumor characterized by frequent mutations/deletions of lysine-specific demethylase 6A(KDM6A),which is suggested to be a key event in cancer progression and metastasis.Beta-elemene has been shown to inhibit metastasis and growth of various tumors,but its effect on KDM6A-null bladder cancer cells remains unknown.Objective:This study aimed to investigate the potential and molecular mechanism ofβ-elemene in inhibiting the growth of KDM6A-null bladder cancer.Methods:This study examined the migration ability and viability of RT-4(KDM6A wild-type)and KU19-19(KDM6A-null)cell lines using wound healing assay and CCK-8,respectively.The inhibitory effect ofβ-elemene on KU19-19 cell migration was evaluated using transwell and immunofluorescence assays,and the expression of transfer-related proteins and genes was analyzed through western blot and qRT-PCR,respectively.Molecular docking was performed to predict the targeting ofβ-elemene,and the effects were confirmed in KDM6Aknockdown RT-4 cells.Finally,the therapeutic effect ofβ-elemene on bladder cancer was tested in animal models.Results:The study observed that loss of KDM6A increased bladder cancer cell migration,with KU19-19 exhibiting significantly stronger migration than RT-4.Further investigation revealed thatβ-elemene effectively inhibited KU19-19 cell migration,likely through targeting EZH2 as determined by molecular docking.Overexpression of KDM6A inhibited KU19-19 metastasis,while knockdown of KDM6A in RT-4 cells enhanced cell migration,which was reversed byβ-elemene treatment.Notably,in vivo testing revealed a significant suppression of KU19-19 cell growth withβ-elemene administered at a dosage of 100 mg/kg.Conclusion:β-elemene has the potential to suppress the growth of KDM6A-null bladder cancer by inhibiting epithelial-mesenchymal transition(EMT),which could make it a promising therapeutic option for patients with KDM6A-null bladder cancer.

PLCD3调控EMT进程促进结直肠癌细胞的增殖、侵袭和迁移

目的:研究磷脂酶PLCD3在结直肠癌组织和细胞中的表达情况和对结直肠癌细胞增殖、侵袭和迁移的影响及发生机制。方法:采用免疫组化分析PLCD3在结直肠癌组织中的表达,利用Kaplan-Meier Plotter数据库得知预后情况。使用RT-qPCR技术验证了PLCD3在人永生化结肠上皮细胞NCM460和结直肠癌细胞的表达水平,利用基因工具干预SW620和SW480细胞。采用克隆形成实验、CCK8增殖实验、划痕实验和Transwell实验来探究PLCD3对结直肠癌细胞增殖、迁移和侵袭的影响。运用Western blot验证EMT相关蛋白的变化。结果:PLCD3在结直肠癌组织中的表达高于癌旁组织(P<0.05),PLCD3高表达与预后生存率低密切相关(P<0.001)。敲低PLCD3抑制了SW620细胞增殖、侵袭和迁移,N-cadherin、MMP2、MMP9相对表达量显著降低(均P<0.01),E-cadherin表达水平显著升高(P<0.001)。过表达PLCD3促进了SW480细胞增殖、侵袭和迁移,N-cadherin、MMP2、MMP9相对表达量显著升高(均P<0.001),E-cadherin表达水平显著降低(P<0.0001)。结论:PLCD3在结直肠癌中表达上调,PLCD3可能通过调控EMT进程促进结直肠癌细胞的增殖、侵袭和迁移。

BMP4通过诱导EMT促进肝癌迁移侵袭

目的:检测BMP4在肝癌中的表达并探讨BMP4在诱导肝癌EMT中的作用,进而研究其对肝癌细胞迁移侵袭能力的影响。方法:采用免疫组织化学方法检测肝癌组织中BMP4的表达,分析其与肝癌临床病理资料之间的关系。将BMP4表达质粒转染至肝癌细胞系Hep G2中,诱导BMP4外源性过表达。观察BMP4转染前、后Hep G2的细胞形态学改变;Western blot检测转染前、后Hep G2中BMP4、EMT相关蛋白(E-cadherin、Vimentin)表达变化情况;划痕和侵袭实验检测BMP4对细胞迁移侵袭能力的影响。结果:BMP4与患者的年龄、病理分级、临床分期、不良预后密切相关。BMP4过表达后Hep G2呈现典型的EMT形态学改变,E-cadherin表达下调、Vimentin表达上调、细胞的迁移侵袭能力显著增强。结论:BMP4与肝癌临床病理资料密切相关,并可能通过诱导EMT促进肝癌细胞的迁移侵袭能力。

CCL20通过AKT/MMP3轴而非EMT途径诱导结肠癌SW480细胞的侵袭和转移

目的:探讨趋化因子CCL20/CCR6促进结肠癌SW480细胞侵袭和迁移的分子机制。方法:筛选高表达CCR6的结肠癌SW480细胞,加入外源性重组人CCL20后,采用Transwell、划痕愈合实验检测其侵袭和迁移能力,以免疫荧光、WB实验检测SW480细胞EMT标志蛋白、AKT信号蛋白以及靶标蛋白MMP3的表达;通过MK2206阻断实验验证AKT信号是其作用机制,通过TCGA数据库资源(https://portal.gdc.cancer.gov/)分析CCL20和MMP3在结直肠癌组织中的表达水平及其相关性。结果:趋化因子CCL20能够明显促进结肠癌SW480细胞侵袭和迁移(均P<0.01),其间并不伴随细胞的EMT变化,而是通过AKT信号的激活及下游靶标蛋白MMP3表达上调是其诱因之一;阻断AKT信号能够明显抑制SW480细胞侵袭和迁移能力,且下调MMP3的表达水平(P<0.05或P<0.01)。TCGA平台数据提示,结肠癌组织中CCL20和MMP3的表达明显高于正常肠黏膜组织,且两者呈明显正相关(r=0.051,P<0.01)。结论:趋化因子CCL20通过AKT/MMP3信号轴而非EMT机制促进结肠癌SW480细胞的侵袭和迁移。

EMT分子标志物在肝癌细胞系中的表达及其意义

目的探讨人肝癌细胞系EMT相关分子标志物的表达情况,为确定相关的肝癌细胞研究模型奠定基础。方法利用实时定量PCR方法检测5株人肝癌细胞系(SNU368、SNU739、HLF、Huh-1和MHCC97L)中E-cadherin、ZO-1、N-cadherin、Vimentin、Snail和Slug的mRNA表达,Westernblot和免疫荧光检测E-cadherin、ZO-1、N-cadherin、Vimentin、Snail和Slug的蛋白表达。结果E-cadherin、ZO-1、N-cadherin、Vimentin、Snail和Slug的mRNA在5种肝癌细胞中均有不同程度表达,E-cadherin、ZO-1、N-cadherin和Slug蛋白在5个细胞株中亦有表达,Vimentin蛋白主要在SNU368、SNU739、HLF和MHCC97L细胞中表达明显,Snail蛋白在SNU368,SNU739,Huh-1和MHCC97L细胞中表达较高。结论HLF细胞系表现出更明显的间质细胞表型特点,Huh-1和SNU368则表现为较典型的上皮细胞表型特点。

EMT研究进展

上皮细胞间质化(EMT)描述了一种从有极性的不能移动的上皮细胞转化为间质细胞的分子重塑和表型改变,在胚胎发育以及成人的损伤修复、组织再生、器官纤维化等方面发挥重要作用,是肿瘤侵袭、转移和肿瘤耐药发生中的一个重要步骤。现在已经发现多个分子通路、miRNAs和EMT有关,由于EMT与肿瘤转移、耐药有重要关系,所以了解EMT相关机制对于了解肿瘤的转移和耐药有重要意义。本文现对EMT相关通路和miRNAs作一综述。

宫颈癌细胞系中EMT相关基因的表达及其意义

目的上皮-间质转化(EMT)是上皮来源的恶性肿瘤侵袭转移的重要机制之一,其特征为上皮细胞标志物表达下调(E-cadherin等)而间质细胞标志物(vimentin等)表达上调。本研究旨在探讨人宫颈癌细胞系EMT相关标志物的表达情况,以确定宫颈癌细胞是否发生EMT,并为确定相关的细胞研究模型奠定基础。方法采用逆转录PCR方法检测4株人宫颈癌细胞系(SiHa、HeLa、RJC-1、CS1213)中E-cadherin、vimentin、Snail和Slug mRNA的表达;Western blotting检测E-cadherin和vimentin的蛋白表达;免疫细胞化学方法检测Snail和Slug的蛋白表达。结果vimentin和Snail mRNA在4种细胞中均有表达,E-cadherin和Slug mRNA在SiHa、HeLa和RJC-1细胞表达。E-cadherin蛋白在SiHa和RJC-1细胞表达,vimentin蛋白在HeLa和CS1213细胞表达。Snail蛋白在4个细胞株都有表达,Slug蛋白在SiHa、HeLa和RJC-1细胞表达。结论 HeLa和CS1213表现出间质细胞表型,SiHa和RJC-1则表现为上皮细胞表型。

肝细胞生长因子(HGF)诱导肝癌Huh7细胞发生上皮间质转化(EMT)后细胞膜表面糖蛋白糖谱的变化

目的应用凝集素芯片技术寻找肝癌细胞表面转移相关的特征性糖谱。方法应用肝细胞生长因子(hepatocyte growth factor,HGF)诱导建立肝癌上皮间质转化(epithelial mesenchymal transition,EMT)细胞模型。通过凝集素芯片比较诱导前后细胞膜的糖谱改变,采用凝集素印迹和荧光细胞凝集素免疫组化方法验证芯片结果。结果诱导后细胞对凝集素ACL、BPL、JAC、MPL、PHA-E、SBA和SNA的亲和作用减弱,而对凝集素AAL、ConA、DBA、GSLⅡ、ECL、HAL、LCA、LTL、NML、NPL、PHA-L、PTLⅡ和WFL的亲和作用增强。提示诱导后肝癌细胞表面出现了黏蛋白T/Tn抗原、平分型N-乙酰葡萄糖胺、α2,6唾液酸和末端α或β连接的N-乙酰半乳糖胺结构减少;而末端和核心岩藻糖、高甘露糖、N-乙酰葡萄糖胺β1,6分支和复杂型寡糖分支结构增多。结论肝癌细胞发生EMT过程中细胞膜表面糖链结构改变,提示糖链结构与肝癌的转移密切相关,为有效控制肝癌转移、改善预后提供了新思路。

肺岩宁方对EMT间质细胞标志因子Vimentin、Fibronectin和N-cadherin表达的影响

目的:研究标志EMT过程发生的间质细胞因子Vimentin、Fibronectin和N-cadherin在C57小鼠Lewis肺癌中的表达,以及肺岩宁方对其mRNA表达的影响。方法:采用Real-TimePCR观察C57小鼠Lewis肺癌右腋下移植瘤和远处转移灶发生的肺组织中Vimentin、Fibronectin和N-cadherin mRNA表达及肺岩宁方对它表达的影响。结果:对于各组移植肿瘤组织中,间质细胞标志因子Vimentin经肺岩宁方治疗后,与模型组相比无差异(P>0.05);而Fibronectin和N-cadherin的表达经肺岩宁方治疗后,与模型组相比明显降低(P<0.01);对于各组肺组织中,Vimentin、Fibronectin和N-cadherin在转移率发生最高的模型组,其表达明显高于正常肺组织中的表达(P<0.01),而经过肺岩宁方治疗后,与模型组相比,Fibronectin和N-cadherin标志因子表达均显著性下降(P<0.05),而Vimentin标志因子治疗前后无差异(P>0.05)。结论:EMT有可能参与了肺癌转移的发生;肺岩宁方具有下调标志EMT过程发生的间质细胞因子Fibronectin和N-cadherin的作用。

OCT4与EMT相关因子在浸润性乳腺癌组织中的表达及其临床意义

目的:观察上皮间质转化(epithelial-mesenchymal transition,EMT)相关因子E-钙黏着蛋白(E-cadherin)和波形蛋白(vimentin)以及干细胞因子八聚体结合转录因子4(octamer-binding transcription factor 4,OCT4)在浸润性乳腺癌中的表达情况及OCT4与E-钙黏着蛋白和波形蛋白的相关性。方法:收集河北医科大学第四医院2009年手术切除的浸润性乳腺癌组织标本40例,实时荧光定量PCR和免疫组织化学法分别检测OCT4、E-钙黏着蛋白和波形蛋白的mRNA和蛋白的表达,分析其与临床病理特征的关系、OCT4和EMT相关蛋白之间的相关性以及与浸润性乳腺癌患者生存率之间的关系。结果:在浸润性乳腺癌组织中,OCT4、E-钙黏着蛋白和波形蛋白表达率分别为30%(12/40)、55%(22/40)和65%(26/40)。OCT4表达与浸润性乳腺癌患者的年龄、组织学分级、淋巴结转移以及Her-2的表达有关,E-钙黏着蛋白表达与淋巴结转移有关,波形蛋白表达与组织学分级和淋巴结转移有关。OCT4 mRNA和蛋白的表达与E-钙黏着蛋白和mRNA蛋白的表达呈负相关,与波形蛋白mRNA和蛋白的表达呈正相关,而E-钙黏着蛋白mRNA和蛋白的表达与波形蛋白mRNA和蛋白的表达也呈负相关。OCT4表达阳性患者的生存率明显低于表达阴性患者,差异具有统计学意义(P<0.05);而E-钙黏着蛋白和波形蛋白的表达与患者生存率之间未见明显相关性(P>0.05)。结论:干细胞因子OCT4可能通过EMT促进浸润性乳腺癌细胞的转移,可能是影响浸润性乳腺癌预后的因素。

基于TGF-β/Smads信号通路探讨胃痞消逆转GPL大鼠胃黏膜EMT效应机制

目的观察健脾化瘀解毒中药复方胃痞消对胃癌前病变(GPL)大鼠TGF-β/Smads信号通路的调控效应,探讨胃痞消逆转GPL大鼠胃黏膜上皮-间质转化(EMT)的效应机制。方法采用N-甲基-N,-硝基-N-亚硝基胍(MNNG)为造模剂结合饥饱失常多因素造模方法复制GPL大鼠模型,连续26周,于16周时开始连续灌胃给药10周,分组如下:正常组,模型组,胃复春组(0.2 g·kg^(-1)),胃痞消高、低剂量(15,7.5 g·kg^(-1))组。观察大鼠胃黏膜组织病理学(HE染色)变化及超微结构;Western Blot法检测尾型同源基因家族2(CDX2)、转化生长因子二型受体(TGF-βRI)I、Rho相关激酶(RhoA)、p-Smad2、p-Smad3、神经钙黏蛋白(N-cad)、β-连环蛋白(β-catenin)表达。结果与正常组比较,模型组GPL大鼠胃黏膜CDX2、RhoA、p-Smad2、p-Smad3和N-cad蛋白表达均显著升高(P<0.05,P<0.01),β-catenin蛋白表达显著降低(P<0.05);与模型组比较,胃痞消高、低剂量组大鼠胃黏膜TGF-βRⅡ、p-Smad2、p-Smad3和RhoA表达均显著降低(P<0.05,P<0.01),胃痞消低剂量组大鼠胃黏膜CDX2表达显著降低(P<0.05),胃痞消高剂量组大鼠胃黏膜N-cad表达显著降低(P<0.01),β-catenin的表达显著升高(P<0.01)。结论胃痞消可拮抗TGF-β/Smads信号通路,进而活化诱导TGF-βRⅡ、p-Smad2、p-Smad3和RhoA表达下调,调控GPL大鼠胃黏膜EMT,延缓GPL向肿瘤的进展。