Impact of lymph node micrometastasis in hilar bile duct carcinoma patients

AIM- To immunohistochemicaUy examine micrometastasis and VEGF-C expression in hilar bile duct carcinoma （HBDC） and to evaluate the clinical significance of the results. METHODS： A total of 361 regional lymph nodes from 25 patients with node-negative HBDC were immunostained with an antibody against cytokeratins 8 and 18 （CAM 5.2）, and immunohistochemical staining of VEGF-C was performed in 34 primary resected tumors. RESULTS： Lymph node micrometastasis was detected in 6 （24%） of the 25 patients and 10 （2.8%） of the 361 lymph nodes. Patients with micrometastasis showed significantly poorer survival rates than those without （P= 0.025）. VEGF-C expression was positive in 17 （50%） of 34 HBDC, and significantly correlated with lymph node metastasis （P=0.042） and microscopic venous invasion （P=0.035）. CONCLUSIONS： It is suggested that immunohistochemically detected lymph node micrometastasis has an impact on the outcome of HBDC. VEGF-C expression is highly correlated with lymph node metastasis in HBDC and might therefore be a useful predictor.

Unexpected metastasis of intraductal papillary neoplasm of the bile duct without an invasive component to the brain and lungs:A case report

BACKGROUND Despite an expanding number of studies on intraductal papillary neoplasm of the bile duct(IPNB),distant metastasis remains unexplained especially in cases of carcinoma in situ.In the present study,we report a rare and interesting case of IPNB without invasive components that later metastasized to lungs and brain.CASE SUMMARY A 69-year-old male was referred to our hospital due to suspected cholangiocarcinoma.Laboratory tests on admission reported a mild elevation of alkaline phosphatase,γ-glutamyl transpeptidase,and total bilirubin in serum.Endoscopic retrograde cholangiography revealed a filling defect in the common bile duct(CBD)extending to the left hepatic duct.Peroral cholangioscopy delineated a tumor in the CBD that had a papillary pattern.Multidetector computed tomography and magnetic resonance cholangiopancreatography detected partial blockage ot interlude in the CBD leading to cholestasis without evidence of metastasis.Therefore,a diagnosis of IPNB cT1N0M0 was established.Left hepatectomy with bile duct reconstruction was performed.Pathological examination confirmed an intraepithelial neoplasia pattern without an invasive component and an R0 resection achievement.The patient was monitored carefully by regular examinations.However,at 32 mo after the operation,a 26 mm tumor in the lungs and a 12 mm lesion in the brain were detected following a suspicious elevated CA 19-9 level.Video-assisted thoracoscopic surgery of left upper lobectomy and stereotactic radiotherapy are indicated.In addition to histopathological results,a genomic profiling analysis using whole exome sequencing subsequently confirmed lung metastasis originating from bile duct cancer.CONCLUSION This case highlights the important role of genomic profiling analysis using whole exome sequencing in identifying the origin of metastasis in patients with IPNB.

High level of urokinase plasminogen activator contributes to cholangiocarcinoma invasion and metastasis

AIM: To investigate the role of urokinase plasminogen activator (uPA) in cholangiocarcinoma (CCA) invasion and its correlation with clinicopathological parameters. METHODS: uPA expression in CCA tissue was determined by immunohistochemistry. The level of uPA from two CCA cell lines (HuCCA-1 and KKU-M213) and a noncancer immortalized cholangiocyte cell line (H69) was monitored by plasminogen-gelatin zymography and western blotting, whereas that of plasminogen activator inhibitor type 1 (PAI-1) protein and uPA receptor (uPAR)mRNA was monitored by western blotting and quantitative real-time reverse transcriptase polymerase chain reaction, respectively. Two independent methods were employed to suppress uPA function: a synthetic uPA inhibitor (B428) and silencing of uPA gene expression using siRNA. In vitro invasion of the uPA-disrupted cells was assessed by Matrigel-coated Transwell assay. RESULTS: The immunohistochemical study showed that 75.3% (131/174) of CCA tissues expressed uPA. High uPA expression was correlated with lymphatic invasion and metastasis of CCA patients. Plasminogen-gelatin zymography of the conditioned media and cell-surface eluates showed that both CCA cell lines, but not H69, expressed both secreted and membrane-bound forms of uPA. Although the two CCA cell lines, HuCCA-1 and KKU-M213, expressed a relatively high level of uPA and uPAR, the latter exhibited a much lower degree of in vitro invasiveness, correlating with a high expression of PAI-1 in the latter, but not in the former. Suppressing uPA function with a specific uPA inhibitor, B428, or with siRNA against uPA reduced in vitro invasiveness of KKU-M213 cells, demonstrating the requirement for uPA in the invasiveness of CCA cells. Therefore, our in vivo and in vitro studies suggest that uPA is an important requirement for the invasion process of CCA. CONCLUSION: uPA expression correlates with lymphatic invasion and metastasis in vivo and is required for CCA cell invasion in vitro , suggesting its potential as a therapeutic target.

Prognostic factors in patients with middle and distal bile duct cancers

AIM: To identify the influence of the surgery type and prognostic factors in middle and distal bile duct cancers.

Multiple von Meyenburg complexes mimicking diffuse liver metastases from esophageal squamous cell carcinoma

Von Meyenburg complexes are benign liver lesions consisting of adenomatous bile duct proliferates. We present two patients suffering from esophageal cancer accompanied by the occurrence of yon Meyenburg complexes. Preoperative computerized tomography （CT） of the liver had not shown these lesions. In one of the patients, diffuse nodular manifestation was found in both liver lobes, mimicking diffuse hepatic metastases. Intraoperative frozen section revealed the benign nature of the lesions in both cases. The patients underwent esophageal resection without complications. To the best of our knowledge, the coincidence of von Meyenburg complexes and esophageal cancer has never been reported before. This uncommon entity should be taken into consideration as a differential diagnosis of liver lesions in malignancies. It underlines the importance of intraoperative frozen section for liver lesions of unknown origin.

Multivariate analysis by Cox Proportional Hazards Model on pro gnoses of patients with bile duct carcinoma after resection

OBJECTIVE: To evaluate the influence of various clinicopathologic factors on the survival of patients with bile duct carcinoma after curative resection. METHODS: A retrospective analysis was performed on 86 cases of bile duct carcinoma treated from January 1981 to September 1995. Fifteen clinicopathologic factors that could possibly influence survival were selected. A multivariate analysis of these individuals was performed using the Cox Proportional Hazards Model. RESULTS: The overall cumulative survival rate was 73% for 1 year, 32% for 3 years and 19% for 5 years. The results of univariate analysis showed that the major significant prognostic factors for influencing survival of these patients were type of histological lesion, lymph node metastasis, pancreatic invasion, duodenal invasion, perineural invasion, macroscopic vessel involvement, resected surgical margin and depth of cancer invasion (P

基于CT图像的纹理分析鉴别肝脏实性局灶性病变

目的 CT是鉴别肝脏实性局灶性病灶的常用检查方法,但其对不典型病灶的鉴别诊断仍有较大的经验依赖性,而纹理分析可以提供客观、定量的图像描述特征。本研究旨在探讨基于CT图像的纹理分析在肝脏实性局灶性病变鉴别诊断中的价值。资料与方法回顾性分析258例经病理证实或临床确诊的肝脏局灶性病变患者的CT图像,其中肝脏局灶性结节增生（FNH）34例,血管瘤（HEM）60例,肝细胞肝癌（HCC）60例,肝内胆管细胞癌（ICC）44例,转移瘤（MET）60例。所有患者均行腹部CT平扫与三期增强扫描。以Ma Zda软件生成CT图像的纹理特征并进行特征筛选,进行各组病灶的判别。结果 258例患者中,基于增强CT图像的纹理分析对于肝脏实性局灶性病变的鉴别诊断错判率（4.26%~37.80%）低于基于平扫图像的纹理分析（9.57%~39.02%）。对于良恶性病变的鉴别,门静脉期图像纹理分析错判率最低（13.57%）;对于FNH与HEM的鉴别,动脉期及门静脉期图像纹理分析效果相当（错判率为4.26%）;对于恶性肿瘤间的鉴别纹理分析错判率相对较高,若于恶性肿瘤间两两鉴别则错判率可降低（错判率最低为HCC与MET,约11.67%）。结论基于CT图像的纹理分析可以作为肝脏实性局灶性病灶鉴别诊断的辅助手段,尤其是FNH与HEM、良性病灶与恶性病灶、恶性病灶间的两两鉴别;其中基于三期增强扫描的纹理分析较基于平扫图像者效果更优。

人胆管癌细胞神经周围浸润模型的构建与初步观察

目的构建人胆管癌细胞神经周围浸润模型,观察培养神经细胞的生长、人胆管癌细胞的增殖与神经周围浸润的过程的形态学变化。方法应用小鼠背根神经节(DorsalRootGanglia,DRG)置于Matrigel基质中体外培养,同时在其表面接种人胆管癌细胞QBC939,观察神经纤维的生长、QBC939细胞粘附、浸润神经纤维的过程及其形态学变化。结果小鼠DRG置于Matrigel基质中体外培养时,神经纤维生长迅速,人胆管癌细胞QBC939能够穿过人工基质粘附并浸润神经纤维,形成包绕神经纤维的细胞团。结论DRG/神经细胞-肿瘤细胞共培养模型较好的模拟了肿瘤细胞穿过细胞基质,浸润神经纤维的过程,人胆管癌细胞QBC939细胞对神经纤维具有亲嗜性。

胆管癌组织中MMP-9的表达及其意义

目的 探讨基质金属蛋白酶 9(matrixmetalloproteinase 9,MMP 9)在胆管癌发展中的作用及与胆管癌浸润、转移和预后的关系。方法 应用免疫组织化学法 (S P法 )对 5 2例经手术和病理证实的胆管癌及 10例经手术切除的正常胆管壁标本进行MMP 9蛋白检测。结果 (1)MMP 9表达在胆管癌组织、癌旁组织及正常胆管上皮组织阳性率分别为 84 6 % (4 4 /5 2 )、2 2 6 % (7/31)和 0 %(0 /10 ) ,有显著性差异 (P <0 0 1)。 (2 )在 5 2例胆管癌组织中 ,MMP 9表达分别与肿瘤的病理分级、TNM分期、浸润和转移及手术方式有关 (P <0 0 5 ) ,而与性别、年龄、肿瘤部位及大小无关 (P >0 0 5 )。 (3)MMP 9表达阳性患者一年生存率明显低于阴性者 (P <0 0 1)。结论 MMP 9表达与胆管癌发展、浸润和转移密切相关 ,检测MMP 9的表达有助于对胆管癌转移潜能和预后的判断。

酪氨酸激酶A和血管内皮生长因子受体2在涎腺腺样囊性癌侵袭转移中的作用

目的通过检测酪氨酸激酶A(TrkA)与血管内皮生长因子受体2(VEGFR2)在涎腺腺样囊性癌(SACC)的表达情况,探讨TrkA与VEGFR2在SACC侵袭转移中的作用。方法采用免疫组织化学SP法检测47例SACC患者组织病理切片中TrkA与VEGFR2的表达情况,结合临床资料统计分析,评价TrkA、VEGFR2与SACC侵袭转移特性的相关性。结果 TrkA、VEGFR2在SACC组织中的阳性率分别是87.23%(41/47例)和85.11%(40/47例),在有神经侵袭、有复发/转移的SACC患者组织切片中TrkA和VEGFR2的表达率均高于无神经侵袭、未复发/转移者,且差别有统计学意义(P<0.05)。组织内微血管密度(MVD)计数与VEGFR2的表达率成正相关关系;MVD在神经侵袭组为25.14±2.83,在无神经侵袭组为18.81±1.33,两者差异有统计学意义(P<0.05);MVD在复发/转移组为26.58±2.38,未复发/转移组为19.06±1.39(P<0.05)。结论 TrkA、VEGFR2的表达强弱与SACC的嗜神经侵袭,复发转移成正相关关系,提示该2种受体在SACC的侵袭转移过程中具有重要作用,并据此推测TrkA、VEGFR2可以作为评价涎腺腺样囊性癌患者预后的指标。

BDNF和TrkB在唾液腺腺样囊性癌中的表达及意义

目的:观察BDNF和TrkB在唾液腺腺样囊性癌组织和细胞系中的表达,揭示BDNF和TrkB与唾液腺腺样囊性癌高转移特性和神经侵袭特性的关系。方法:以30例SACC标本为研究对象,以5例正常腮腺及3例腺泡细胞癌作为对照,采用免疫组化法对BDNF和TrkB进行检测。以SACC高、低转移细胞系ACC-2和ACC-M为研究对象,利用免疫组化、RT-PCR、实时荧光定量PCR和Western印迹,检测BDNF和TrkB在其中的表达差异。采用χ2检验进行统计学分析。结果:组织标本免疫组化显示:TrkB表达水平在嗜神经现象组显著高于未见嗜神经现象组(Ρ<0.05),BDNF表达水平与嗜神经现象无关(Ρ>0.05)。实时荧光定量PCR和Western印迹均检测到BDNF和TrkB在ACC-M中的表达高于ACC-2(Ρ<0.01)。结论:TrkB的表达强度与SACC的神经侵袭特性相关,BDNF和TrkB的表达强度与SACC的高转移特性相关。

涎腺腺样囊性癌生物学特性与生物治疗

腺样囊性癌具有两大生物学特性,即嗜神经侵袭和肺高转移特性。这两大生物学特性直接影响腺样囊性癌的治疗和预后,因而一直受到国内外学者的广泛关注,并作了大量的研究,取得了一系列进展。腺样囊性癌对于放疗、化疗均不敏感,目前仍以手术治疗为主。近年来,随着肿瘤生物治疗的迅速发展,国内外学者探索将生物治疗运用于腺样囊性癌,并取得了有益的成果。

NDRG1在乳腺浸润性导管癌中的表达及其与侵袭、转移的关系

目的研究NDRG1(N-myc downstream regulated gene 1)基因在乳腺浸润性导管癌中的表达,及其与乳腺癌侵袭、转移的关系。方法应用免疫组化法检测26例伴淋巴结转移的乳腺浸润性导管癌和45例无淋巴结转移的乳腺癌中的NDRG1、MMP2、MMP9、TIMP2、E-cad的表达状态。结果乳腺癌伴淋巴结转移的标本中NDRG1的表达率低于无淋巴结转移标本(P<0.01)。NDRG1蛋白的表达与TIMP2的表达正相关(P<0.05);而与MMP2、MMP9和E-cad的表达无关(P>0.05)。结论NDRG1与浸润性导管癌的侵袭和转移能力密切相关,可能是一个候选的肿瘤转移抑制基因,并且可能成为乳腺癌的一个有用的预后指标。

法尼酯X受体FXR在乳腺浸润性导管癌中的表达

目的探讨法尼酯X受体(FXR)在乳腺浸润性导管癌组织中的表达。方法用免疫组织化学MaxVision法检测FXR、雌激素ER、孕激素PR在10例正常乳腺组织和34例乳腺浸润性导管癌组织中的表达。结果 FXR在正常乳腺组中阳性率100%,浸润性导管癌中阳性率55.88%。FXR蛋白表达与年龄、病理分级无明显关系(P>0.05),与淋巴结未转移组比较,有淋巴结转移组高表达FXR蛋白(P<0.05)。FXR蛋白与ER蛋白表达呈显著正相关(r=0.373,P<0.05);PR蛋白高表达时,虽FXR蛋白也高表达,但无相关性(r=0.206,P>0.05)。结论 FXR高表达可能促进乳腺癌淋巴结转移,FXR与ER雌激素表达呈显著正相关。

反义寡核苷酸转染对胆管癌细胞侵袭能力的实验研究

目的 探讨层粘连蛋白受体反义寡核苷酸 (AS OD)对胆管癌细胞体外侵袭和转移能力的影响。方法 以层粘连蛋白受体反义寡核苷酸片段转染胆管癌细胞高转移性亚群 ,RT PCR和流式细胞仪检测转染前后细胞层粘连蛋白受体mRNA和蛋白的表达差异 ,体外粘附实验和侵袭实验检测转染前后细胞粘附和侵袭能力的改变。结果 反义寡核苷酸转染后可在mRNA水平和蛋白水平下调层粘连蛋白受体的表达 ,且下调作用呈剂量依赖性。与空白对照组及顺义寡核苷酸转染组比较相差均显著 (P<0 0 5 )。体外粘附和侵袭实验显示转染后细胞粘附和侵袭能力均较之转染前有显著下降 ,两者间相差均显著 (P <0 0 5 )。结论 层粘连蛋白受体反义寡核苷酸可降低胆管癌细胞体外侵袭和转移能力。

整合素α_Vβ_3在人胆管癌组织中的表达及其临床意义

目的 研究整合素αVβ3在人胆管癌组织中的表达及其与胆管癌的临床病理关系。方法 采用免疫组织化学方法检测47例人胆管癌组织及正常癌旁胆管组织中整合素αVβ3的表达。结果 整合素αVβ3表达于人胆管癌细胞及胆管癌组织中的新生血管细胞,47例标本均呈阳性表达,其表达强度与是否伴有肿瘤转移有显著差异(P <0 0 5 ) ,而与性别、癌组织分化程度无关。结论 整合素αVβ3可能在胆管癌细胞的恶性生物学行为中具有重要的作用,检测整合素αVβ3表达可以作为胆管癌预后判断的依据之一。

胆囊收缩素对人胆管癌细胞粘附、运动及侵袭能力的影响

目的研究胆囊收缩素对胆管癌细胞的粘附、运动以及侵袭行为的影响。方法体外培养人胆管癌细胞系QBC939,应用粘附试验、Transwell趋化运动和侵袭模型,研究CCK8S及其不同浓度下对胆管癌细胞的粘附、运动以及侵袭能力的影响。结果CCK8S能够以浓度依赖方式降低QBC939细胞的粘附,而促进其运动和侵袭能力,其受体拮抗剂L18和L60能够明显减弱CCK8S抑制QBC939细胞粘附的作用以及促进瘤细胞运动和侵袭的能力,细胞运动和侵袭实验显示L60具有较强的拮抗CCK8S作用的能力。结论CCK8S具有降低QBC939细胞的粘附、促进QBC939运动和侵袭的能力,CCK2受体在影响QBC939细胞的运动、侵袭能力方面可能具有较强的作用。

胃癌侵犯血管、神经及其临床意义

目的 探讨胃癌侵犯血管、神经及其对胃癌侵袭、转移和预后的意义。方法 对 6 8例进展期胃癌标本次连续切片进行血管内膜下弹力纤维VB :HE组织化学染色以及S - 10 0、CD34免疫组化染色 ,进行镜下观察和单因素及多因素分析。结果 血管受侵 52例 ( 76 .4 % ) ,表现为 4种类型 ,即血管壁受侵、游离癌细胞侵入血管腔、部分栓塞和完全栓塞。神经受侵犯 2 2例( 32 .35% )。血管、神经侵犯与肿瘤大小、侵袭深度、淋巴结转移和肿瘤TNM分期显著相关。单因素预后分析表明 ,有血管、神经侵犯的胃癌患者较无血管、神经侵犯者 ,预后明显较差。但多因素分析表明 ,胃癌对血管、神经的侵犯不是影响预后的独立因素。结论 血管和神经间隙均是胃癌侵袭转移的重要途径 ,胃癌对血管、神经的侵犯在一定程度上反映了癌细胞的侵袭能力 ,了解胃癌对血管、神经的侵袭情况对判断干预有一定的参考价值。

吡格列酮对肝门部胆管癌QBC939细胞增殖、凋亡和侵袭力的影响

目的:探讨过氧化物酶体增殖物激活受体-γ(PPAR-γ)配体吡格列酮(pioglitazone,PGZ)对胆管癌细胞增殖、凋亡和体外侵袭力的影响.方法:体外培养人肝门胆管癌QBC939细胞;采用流式细胞技术与TUNEL酶标记法检测PGZ对QBC939细胞增殖和凋亡的影响.通过Matrigel侵袭实验和过河实验检测PGZ对QBC939细胞体外侵袭力和运动能力的影响.结果:流式细胞仪分析显示,PGZ能明显抑制QBC939细胞增殖,使其停滞于G2/M期,促使诱导细胞凋亡;TUNEL酶标记显示凋亡的QBC939细胞出现凋亡小体;Matri-gel侵袭实验和过河实验显示,随着PGZ浓度的增加,透过Matrigel膜的细胞数减少,过河时间延长(P<0.01),呈剂量-效应关系.结论:PGZ能明显抑制QBC939细胞增殖,诱导细胞凋亡,对QBC939细胞的体外侵袭力有明显的抑制作用.PPAR-γ配体PGZ有望成为治疗胆管癌新的切入点.

OPN在胆管腺癌组织中的表达及其与预后的关系

目的:检测OPN在胆管腺癌、癌旁胆管组织中的表达,了解OPN在胆管腺癌组织中的表达及其与预后的关系。方法:采用免疫组化SP法检测OPN在44例胆管腺癌、44例癌旁胆管组织中的表达情况,分析OPN与胆管癌临床病理参数的关系,并对患者进行随访。结果:OPN在胆管腺癌、癌旁胆管组织中表达阳性率分别为68.2%(30/44)、27.3%(12/44),在胆管腺癌组织中表达阳性率明显高于癌旁胆管组织(P<0.05)。OPN在胆管腺癌组织中的表达与胆管癌浸润深度、淋巴结转移相关(P<0.05),差异有统计学意义。OPN阳性表达患者术后5年总体生存率明显低于阴性患者(P=0.015)。结论:OPN在胆管腺癌组织中高表达,与胆管癌的侵袭、转移及预后密切相关,可作为胆管腺癌的参考指标,对胆管腺癌的诊断和预后评估具有重要意义。