Livin表达与胃癌上皮-间质转化的关系及临床意义

目的探讨Livin表达与胃癌上皮-间质转化(epithelial-mesenchymal transition,EMT)的关系及临床意义。方法采用免疫组化法检测110例胃癌及相应癌旁正常胃组织中Livin和EMT标志蛋白的表达,分析Livin表达与胃癌临床病理特征的关系及Livin表达与EMT标志蛋白表达的相关性。应用Western blot法检测8例新鲜胃癌组织中Livin和EMT标志蛋白的表达。结果胃癌组织中Livin阳性率(59. 1%)明显高于相应癌旁组织(8. 2%)(P 0. 05)。Livin表达与胃癌淋巴结转移相关(P 0. 001)。Western blot检测结果显示Livin在有淋巴结转移的胃癌组中表达高于无淋巴结转移组。胃癌组织中E-cadherin、vimentin和N-cadherin的阳性率分别为29. 1%、30. 9%和57. 3%。胃癌组织中Livin与E-cadherin的表达呈负相关(r=-0. 322,P=0. 001),与vimentin和N-cadherin的表达呈正相关(r=0. 276,P=0. 004; r=0. 216,P=0. 024)。Livin高表达的胃癌组织中,vimentin和N-cadherin亦高表达,而E-cadherin表达降低,反之亦然。结论 Livin表达与胃癌组织的EMT表型有关,并促进胃癌的转移。     

RASSF7表达与乳腺癌上皮-间质转化的关系

目的探讨RASSF7在乳腺癌中的表达及与乳腺癌上皮-间质转化(epithelial-mesenchymal transition,EMT)的关系。方法采用免疫组化法检测80对乳腺癌及癌旁正常乳腺组织中RASSF7的表达,分析RASSF7表达与乳腺癌临床病理特征的关系。采用Western blot法检测8对新鲜乳腺癌及癌旁正常乳腺组织中RASSF7的表达。Western blot法检测在低氧条件下MDA-MB-231细胞中RASSF7、HIF-1α、EMT相关标志蛋白(E-cadherin、vimentin和N-cadherin)的表达水平。进一步分析乳腺癌组织中RASSF7的表达与HIF-1α及EMT相关标志蛋白(E-cadherin、vimentin和N-cadherin)表达的相关性。结果乳腺癌组织中RASSF7的阳性率(70.0%,56/80)明显高于癌旁正常乳腺组织(27.5%,22/80)(P<0.05),Western blot法结果亦证实,乳腺癌组织中RASSF7蛋白表达高于癌旁正常乳腺组织。RASSF7表达与乳腺癌淋巴结转移和TNM分期呈正相关(P=0.002,P=0.027)。低氧组RASSF7、HIF-1α、vimentin和N-cadherin的蛋白表达水平高于常氧组,E-cadherin在低氧组表达低于常氧组。进一步统计分析结果表明,乳腺癌组织中RASSF7表达与HIF-1α、vimentin和N-cadherin表达呈正相关(r=0.245,P=0.029;r=0.237,P=0.034;r=0.301,P=0.007),与E-cadherin的表达呈负相关(r=-0.33,P=0.003)。结论低氧环境下RASSF7可能通过引起乳腺癌EMT,促进乳腺癌的侵袭、转移。     

子宫颈鳞状细胞癌中GDF-15与EMT相关蛋白的表达及相关性

目的检测子宫颈鳞状细胞癌组织中GDF-15及上皮-间质转化(epithelial-mesenchymal transition, EMT)相关蛋白E-cadherin、N-cadherin、vimentin的表达,探讨GDF-15在子宫颈鳞状细胞癌发生、侵袭中的生物学作用。方法采用免疫组化EliVision两步法检测85例子宫颈鳞状细胞癌组织及其配对癌旁子宫颈组织中GDF-15、E-cadherin、N-cadherin、vimentin的表达,分析GDF-15表达与子宫颈鳞状细胞癌临床病理特征的关系及GDF-15与EMT相关蛋白表达的相关性。结果与癌旁子宫颈组织相比,子宫颈鳞状细胞癌组织中GDF-15、N-cadherin和vimentin表达上调,E-cadherin表达下调,差异有统计学意义(P均0.05)。子宫颈鳞状细胞癌组织中GDF-15表达与N-cadherin和vimentin表达呈正相关(r_s=0.422,r_s=0.374),与E-cadherin表达呈负相关(r_s=-0.305)。结论 GDF-15与EMT相关蛋白在子宫颈鳞状细胞癌组织中异常表达,过表达GDF-15在子宫颈癌的发生、侵袭中发挥重要作用,可能与诱导子宫颈癌EMT有关。     

乳腺癌中转录因子DLX4与EMT相关蛋白的表达及意义

目的 探讨转录因子DLX4与上皮细胞-间质转化(epithelial-mesenchymal transition,EMT)相关蛋白在乳腺癌组织和乳腺细胞系中的表达及其相关性。方法 收集100例乳腺癌及对应癌旁组织标本,采用免疫组化SP法和Western blot法检测乳腺癌组织和乳腺细胞系中DLX4与EMT相关蛋白E-cadherin和vimentin的表达,并分析三者与乳腺癌临床病理特征的关系。结果 乳腺癌组织中DLX4和vimentin阳性率(68.0%和53.0%)显著高于癌旁组织(22.0%和28.0%)(P0.05),乳腺癌组织中E-cadherin阳性率(32.0%)显著低于癌旁组织(76.0%)(P0.05)。Western blot法检测结果显示,在高侵袭性乳腺癌细胞系中DLX4、vimentin均呈高表达,E-cadherin呈低表达;DLX4、E-cadherin及vimentin的表达与乳腺癌TNM分期和淋巴结转移相关。Spearman相关分析结果显示,乳腺癌中DLX4与E-cadherin表达呈负相关(r_s=-0.505,P=0.000),DLX4与vimentin表达呈正相关(r_s=0.165,P=0.016)。结论 乳腺癌中转录因子DLX4与E-cadherin表达呈负相关,与vimentin表达呈正相关,可能存在调控关系,从而促进乳腺癌的局部侵袭和远处转移。     

Brachyury在非小细胞肺癌高表达并与EMT及患者的不良预后相关

目的研究非小细胞肺癌中Brachyury表达的变化及其与患者预后的关系。方法收集115例NSCLC患者的标本及30例癌旁正常肺组织,应用免疫组织化学的方法观察Brachyury在非小细胞肺癌中的变化及其与EMT标志物E-cadherin及N-cadherin的关系,应用Western Blot结合RT-PCR方法检测肺癌组织中Brachyury蛋白与mRNA水平的变化结果 Brachyury在非小细胞肺癌中表达明显增加Brachyury的表达水平与Ecadherin呈正相关,但与N-cadherin呈负相关,Brachyury在非小细胞肺癌中的高表达与患者的不良预后密切相关。结论 Brachyury的表达与非小细胞肺癌的发生发展紧密相关。     

PDCD5通过Wnt/β-catenin通路抑制肾细胞癌侵袭转移及其机制研究

目的探讨肾细胞癌(RCC)患者PDCD5的表达水平及其参与调控RCC侵袭与转移的可能机制。方法利用免疫组化检测RCC标本与癌旁组织中PDCD5的蛋白表达水平,同时分析PDCD5与疾病分期、预后的相关性;利用transwell及划痕实验检测PDCD5对肾癌细胞系A498侵袭与转移的影响;利用Western blot检测A498细胞转染PDCD5后MMP2、MMP9、E-cadherin、N-cadherin及Wnt/β-catenin通路的变化。结果 (1)与癌旁组织相比,RCC标本中PDCD5明显下调(P0.001),且与疾病分期及预后正相关;(2)PDCD5明显抑制了肾癌细胞系A498的转移与侵袭能力(P0.01);(3)PDCD5通过抑制A498细胞系中MMP2、MMP9及N-cadherin的表达水平,增强E-cadherin的表达水平调控EMT转化,同时Wnt/β-catenin通路关键蛋白下调。结论 PDCD5通过调控Wnt/β-catenin通路影响肾细胞癌EMT转化,负性调控了RCC病人的侵袭与转移进程。     

趋化因子SDF-1调控结直肠癌细胞上皮-间质转化

目的通过检测Twist、SDF-1、E-cadherin在结直肠癌及癌旁组织中的表达,探讨三者与上皮-间质转化(epithelial-mesenchymal transition,EMT)的相关性,为肿瘤的分子靶向治疗提供可能的靶标。方法收集结直肠癌和对应癌旁正常新鲜标本组织40例和结直肠癌组织及对应癌旁组织蜡块90例,利用免疫组化SP法和qRT-PCR法检测结直肠癌组织中SDF-1及EMT相关因子E-cadherin、Twist的表达,统计学分析SDF-1表达与E-cadherin、Twist表达之间的相关性。结果 qRT-PCR结果显示,与对应癌旁组织相比,结直肠癌组织中SDF-1、Twist mRNA的表达水平明显升高,而Ecadherin mRNA表达水平显著下降,差异有统计学意义(P0.05);结直肠癌组织中SDF-1、Twist的阳性率分别为75.56%、62.22%,明显高于对应癌旁组织,而E-cadherin在肿瘤中的阳性率为28.89%,显著低于癌旁组织,差异有统计学意义(P0.05)。SDF-1表达与E-cadherin表达呈负相关,而与Twist表达呈正相关。结论趋化因子SDF-1高表达可能参与结直肠癌细胞的EMT过程,Twist高表达和Ecadherin低表达提示EMT现象参与结直肠癌的形成及转移过程,并促进肿瘤的生成和转移。     

IL-8在胃癌组织上皮间质转化过程中表达增强

目的研究胃癌上皮间质转化(EMT)过程中趋化因子IL-8的调节机制与意义。方法用Western blot、酶联免疫吸附试验(ELISA)等技术,检测胃癌组织与正常组织中IL-8以及EMT相关基因(E-cadherin, vimentin, twist等)的表达,胃癌细胞系MGC-803对IL-8与胃癌细胞EMT的相关性进行分析和探讨。结果肿瘤组织中IL-8的表达水平(710.5±89.9)显著高于癌旁组织(510.5±85.5)(P0.001)。与正常组织相比,胃癌组织中IL-6、twist及vimentin表达水平明显高于正常胃黏膜组织(P0.05),而E-cadherin表达水平低于正常组织(P0.05)。IL-8因子的分泌与twist蛋白(P0.001)和vimentin蛋白(r=0.454,P0.001)表达水平呈显著正相关性,与E-cadherin蛋白表达水平呈显著负相关性(P0.001)。结论胃癌组织中趋化因子IL-8的表达量增强, IL-8与胃癌侵袭转移相关,同时IL-8的表达量也可作为胃癌侵袭转移患者预后判断的参考标准之一。     

敲低核仁小RNA宿主基因6(SNHG6)抑制舌癌细胞的增殖及上皮间质转化

目的探究长链非编码RNA核仁小RNA宿主基因6 (SNHG6)在舌鳞状细胞癌组织、Tca1183舌癌细胞中的表达及对舌癌细胞生物学特性的影响。方法使用实时定量PCR检测舌鳞状细胞癌组织与正常组织、Tca1183舌癌细胞与正常舌上皮细胞中SNHG6的mRNA水平,分析舌癌临床病理指标与SNHG6表达的相关性。构建SNHG6干扰质粒并转染至Tca1183舌癌细胞,噻唑蓝(MTT)法检测细胞活力,集落形成实验检测细胞增殖能力,流式细胞术检测舌癌细胞凋亡变化; Western blot法检测细胞上皮间质转化(EMT)相关蛋白β联蛋白(β-catenin)、上皮钙黏素(E-cadherin)、神经钙黏素(N-cadherin)和波形蛋白(vimentin)的蛋白水平。结果与正常组织和正常舌上皮细胞相比,舌癌组织以及舌癌细胞中SNHG6的mRNA水平显著升高。舌癌组织SNHG6 mRNA水平与患者年龄和性别等不相关,与组织分化差、临床分期高和淋巴结转移呈正相关。干扰SNHG6在Tca1183舌癌细胞中的表达能够抑制癌细胞增殖以及促进细胞凋亡,且可以通过促进β-catenin和E-cadherin蛋白表达以及抑制N-cadherin和vimentin蛋白表达抑制Tca1183细胞EMT。结论 SNHG6在舌癌组织中高表达,敲低舌癌细胞中SNHG6能够抑制其增殖和EMT。     

结肠癌组织中LKB1的表达及其与上皮-间质转化的关系

目的探讨LKB1在结肠癌组织中的表达及其与肿瘤上皮-间质转化的关系。方法采用免疫组化法检测75例结肠癌组织及对应癌旁正常结肠组织中LKB1、上皮-间质转化(epithelial mesenchymal transformation,EMT)相关标志物E-cadherin、N-cadherin、vimentin的表达,分析LKB1表达与结肠癌临床病理特征及EMT的关系。结果 49例结肠癌组织中LKB1低表达,55例正常结肠组织中LKB1高表达;LKB1表达降低与淋巴结转移、TNM肿瘤分期相关(P0.05);E-cadherin、N-cadherin、vimentin分别在25例、48例、26例结肠癌组织中高表达,LKB1表达与E-cadherin表达呈正相关(r=0.648,P0.05),与Ncadherin(r=-0.891,P0.05)和vimentin(r=-0.660,P0.05)表达呈负相关。结论 LKB1在结肠癌组织中表达下调,与EMT的发生呈正相关,可作为结肠癌转移发生的预测指标。     

Differential expression of the epithelial-mesenchymal transition regulators snail,SIP1, and twist in gastric cancer

Epithelial-mesenchymal transition (EMT) involving down-regulation of E-cadherin is thought to play a fundamental role during early steps of invasion and metastasis of carcinoma cells. The aim of our study was to elucidate the role of EMT regulators Snail, SIP1 (both are direct repressors of E-cadherin), and Twist (an activator of N-cadherin during Drosophila embryogenesis), in primary human gastric cancers. Expression of Snail, SIP1, and Twist was analyzed in 48 gastric carcinomas by real-time quantitative RT-PCR in paraffin-embedded and formalin-fixed tissues. The changes of expression levels of these genes in malignant tissues compared to matched non-tumorous tissues were correlated with the expression of E- and N-cadherin. From 28 diffuse-type gastric carcinomas analyzed reduced E-cadherin expression was detected in 11 (39%) cases compared to non-tumorous tissues. Up-regulated Snail could be found in 6 cases with reduced or negative E-cadherin expression. However, there was no correlation to increased SIP1 expression. Interestingly, we could detect abnormal expression of N-cadherin mRNA in 6 cases, which was correlated with Twist overexpression in 4 cases. From 20 intestinal-type gastric cancer samples reduced E-cadherin expression was found in 12 (60%) cases, which was correlated to up-regulation of SIP1, since 10 of these 12 cases showed elevated mRNA levels, whereas Snail, Twist, and N-cadherin were not up-regulated. We present the first study investigating the role of EMT regulators in human gastric cancer and provide evidence that an increase in Snail mRNA expression is associated with down-regulation of E-cadherin in diffuse-type gastric cancer. We detected abnormally positive or increased N-cadherin mRNA levels in the same tumors, probably due to overexpression of Twist. SIP1 overexpression could not be linked to down-regulated E-cadherin in diffuse-type tumors, but was found to be involved in the pathogenesis of intestinal-type gastric carcinoma. We conclude that EMT regulators play different roles in gastric carcinogenesis depending on the histological subtype.     

Clinical implications of N-cadherin expression in gastric cancer

Neo-expression of N-cadherin in cancer cells is regarded as a significant event in tumor progression via epithelial-mesenchymal transition (EMT). No reports have detailed the clinical impact of N-cadherin expression in gastric cancer. We retrospectively examined the co-expression of N-cadherin and E-cadherin in human gastric carcinoma and analyzed the clinicopathological significance of N-cadherin expression. One hundred and forty-six gastric cancer patients who received curative gastrectomy were enrolled. E-cadherin and N-cadherin immunoreactivity in cancer tissue was evaluated by the avidin-biotin-peroxidase complex technique. The correlation between N-cadherin and E-cadherin expression and clinicopathological parameters were analyzed. N-cadherin-positive and -negative expression were found in 31 and 115 patients, respectively. N-cadherin expression positively correlated with hematogenous recurrence (P < 0.01) and negatively correlated with patients' postoperative outcomes (P < 0.05). Moreover, only in the E-cadherin-preserved group was prognostic significance found according to N-cadherin expression (P < 0.01). We could not show a significant relationship between N-cadherin expression and EMT in gastric cancer. However, neo N-cadherin expression significantly affected patient's survival in gastric cancer. Therefore, we concluded that neo N-cadherin expression may be a useful prognostic marker independent of E-cadherin expression.     

结合恶性胸腔积液中脱落细胞探讨上皮细胞-间质转化与非小细胞肺癌的相关性

目的探讨恶性胸腔积液(malignant pleural effusion,MPE)中非小细胞肺癌(non-small cell lung cancer,NSCLC)上皮细胞-间质转化(epithelial-mesenchymal transition,EMT)相关分子标志物的表达及意义。方法对823例胸水标本进行液基细胞薄层检测(thinPrep cytology test,TCT),结合活检结果确诊107例NSCLC标本,随机分成NSCLC细胞组、NSCLC活检组及NSCLC癌旁组织组三组,对其中39例NSCLC细胞组胸水标本进行沉渣后包埋、切片,采用免疫组化SP法检测三组中vimentin、E-cadherin、N-cadherin、β-catentin的表达,并进行对比分析。结果E-cadherin、β-catentin阳性/异常阳性率在NSCLC细胞组、活检组均明显低于癌旁组织组(P<0.05)。NSCLC细胞组、活检组N-cadherin、vimentin的阳性率均明显高于癌旁组织组(P<0.05)。同时亦发现,在NSCLC细胞组中,E-cadherin表达与分化程度和淋巴结转移有关(P<0.05),β-catentin异常阳性与分化程度有关(P<0.05),N-cadherin与vimentin表达与淋巴结转移有关(P<0.05)。以上4个抗体表达/异常表达均与患者年龄、性别、吸烟史和组织学分型无关(P>0.05)。结论MPE中NSCLC细胞具备EMT表型,从细胞学角度进一步证实EMT现象存在于NSCLC,为新的靶向治疗药物的研究和治疗靶点提供新思路。     

E-cadherin和FOXO3a在胃癌组织和细胞中表达的关系

目的:探讨上皮钙黏素(E-cadherin)和叉头框蛋白O3a(FOXO3a)在胃癌组织和细胞中表达的关系及意义。方法:应用免疫组织化学法检测53例胃癌组织及对应癌旁组织中E-cadherin和FOXO3a的表达情况,分析两者的表达水平及与临床病理参数的关系。构建E-cadherin过表达的胃癌稳转细胞株AGS,免疫细胞化学法检测E-cadherin及FOXO3a蛋白表达,Western blot法检测细胞中E-cadherin、FOXO3a、Akt、Bcl-2和Bax的蛋白表达,CCK-8法检测细胞活力。结果:胃癌组织中E-cadherin和FOXO3a的阳性率较对应癌旁组织均显著降低(P0.05)。E-cadherin表达与胃癌分化程度和TNM分期显著相关(P0.05),与年龄、性别、肿瘤部位、T分期及淋巴结转移无显著相关。FOXO3a表达与胃癌分化程度显著相关(P0.05),与年龄、性别、部位、TNM分期、T期及淋巴结转移无显著相关。胃癌组织中E-cadherin与FOXO3a表达存在显著正相关(r=0.376,P=0.003)。E-cadherin过表达后,胃癌AGS细胞活力显著下降,细胞中FOXO3a和Bax表达显著升高,Akt表达显著下降。结论:E-cadherin与FOXO3a共同参与胃癌的发生发展,E-cadherin可能通过调节Akt/FOXO3a信号传导影响胃癌细胞活力。     

Effect of eribulin on epithelial–mesenchymal transition plasticity in metastatic breast cancer: An exploratory,prospective study

Epithelial–mesenchymal transition (EMT) plays a pivotal role in cancer metastasis and treatment resistance, which worsens prognosis. In phase III trials, eribulin improved overall survival in metastatic breast cancer (MBC) patients. In preclinical studies, eribulin suppressed EMT. However, clinical data on the use of eribulin for MBC patients are limited. In this exploratory, prospective study, we examined the effect of eribulin on EMT in MBC patients. Twenty-two patients aged 44–82 years with recurrent breast cancer or MBC were treated with eribulin. Breast cancer tissue samples were obtained before treatment and on Day 15 ± 5 of the first cycle of eribulin treatment. EMT markers (E-cadherin, claudin-3, vimentin, and N-cadherin) were analyzed using western blotting. EMT changes were evaluated based on the ratio of epithelial to mesenchymal markers before and after treatment in individual tumors. E-cadherin/vimentin, claudin-3/vimentin, E-cadherin/N-cadherin, and claudin-3/N-cadherin ratios were significantly higher after treatment (p = .007, p = .005, p = .006, and p = .011, respectively). Based on E-cadherin/vimentin, 65.0% of tumors shifted to an epithelial phenotype, as compared to 66.7% based on claudin-3/vimentin, 84.6% based on E-cadherin/N-cadherin, and 71.4% based on claudin-3/N-cadherin ratios. Thus, our results showed that eribulin suppressed EMT in breast cancer tissues.     

Up-regulated LINC00261 predicts a poor prognosis and promotes a metastasis by EMT process in cholangiocarcinoma

ObjectiveLINC00261 plays a vital role in tumorigenesis and metastasis of digestive system cancer. However, an influence of LINC00261 on cholangiocarcinoma has a little research. There, we investigated clinical role and molecular mechanisms of LINC00261 in cholangiocarcinoma.MethodsThe qRT-PCR was performed for the detection of LINC00261 level in 50 paired specimens from CCA patients and six cell lines. Cell proliferation were explored by CCK-8 and colony formation assays in QBC939 and RBE cells after transfected with si-LINC00261 or si-NC. Then, AO/EB double fluorescence staining and flow cytometric assays were performed to assess cell apoptosis. Transwell and wound healing assays were selected to evaluate migratory and invasive property of cells. Protein levels, such as PCNA, Bax, Bcl-2, and several epithelial-to-mesenchymal transition markers, including E-cadherin, N-cadherin and Vimentin, were detected by western blot assays. Furthermore, we use a R2 platform to evaluate the correlation between LINC00261 and EMT makers and predict the overall survival and relapse-free survival for CCA patients by the expression of LINC00261/ EMT makers.ResultsLINC00261 was overexpressed in cancerous tissues and CCA cell lines compared with adjacent tissues and HIBEC, respectively. Up-regulation of LINC00261 was related to larger tumor size (p = 0.009), positive lymph node metastasis (p = 0.021), advanced TNM stages (p = 0.017) and higher postoperative recurrence (p = 0.009) for CCA patients. Additionally, univariate and multivariate analysis displayed that LINC00261 an independent prognostic factor in CCA patients. Knockdown of LINC00261 expression in RBE and QBC939 cell lines inhibited cell proliferation, migration and invasion property and increased cell apoptosis and the EMT progression. Moreover, there was a strong correlation between LINC00261 and E-cadherin (CDH1) (p < 0.05), and low expression of E-cadherin (CDH1) has a poor overall survival and relapse-free survival in CCA patients (p < 0.05).ConclusionOverall, high level of LINC00261 in CCA predicts a poor prognosis, and promotes a metastasis via EMT process. Thus, LINC00261 could be a promising biomarker and therapeutic target for CCA, and in the high level of LINC00261 in CCA, E-cadherin or CDH1 might be an effective factor for tumor metastasis or poor prognosis.     

Endophilin A2与胃癌BGC-823细胞转移及上皮-间质转化的关系

目的:探究Endophilin A2在胃癌转移及上皮-间质转化(epithelial-mesenchymal transition,EMT)过程中的作用及相关机制.方法:通过免疫组织化学法测定37例胃癌原发灶组织和35例胃癌转移淋巴结中Endophilin A2蛋白的分布和表达情况.在BGC-823细胞转染腺病毒过表达EndophilinA2后,通过划痕实验和Transwell实验观察其迁移能力;Western印迹检测EMT及相关信号通路蛋白的水平.结果:免疫组织化学结果显示胃癌淋巴结转移灶中Endophilin A2蛋白的表达明显低于原发灶.BGC-823细胞中过表达Endophilin A2之后细胞的迁移能力降低,上皮细胞标志物上皮黏附蛋白(epithelium cadherin,E-cadherin)的表达升高,同时黏附蛋白辅助因子p-catenin的表达升高,而间质细胞标志物神经钙黏蛋白(nervous cadherin,N-cadherin)和波形蛋白vimentin的表达明显下降.Endophilin A2过表达之后,Notch和snail的表达也明显降低.结论:Endophilin A2与胃癌淋巴转移密切相关,过表达Endophilin A2可能通过Notch/snail通路抑制胃癌细胞BGC-823的EMT进程.