胆管细胞癌酪氨酸激酶激活及其磷酸化酶分布

目的:探讨胆管细胞癌组织酪氨酸磷酸化信号通路变化及相关分子情况,以寻找新的治疗靶点。方法:免疫亲和、LC-MS/MS分析识辨胆管癌病人(n=23)750多种不同蛋白质中1 000多个酪氨酸磷酸化位点。结果:胆管细胞癌在DDR1、EPHA2、EGFR和ROS1表现最高水平酪氨酸激酶磷酸化,高表达DDR1、EPHA2、ROS1酪氨酸激酶活性(n=18,78%),低表达或不表达RTK活性(n=5,22%);癌旁组织EGFR、AXL、EPHB4和PDGFRA表现出最高水平酪氨酸磷酸化;肿瘤组织和癌旁组织磷酸化蛋白种类分布类似。结论:胆管细胞癌组织中酪氨酸激酶激活,DDR1、EPHA2、EGFR和ROS1酪氨酸激酶活性比癌旁组织高表达,可能致胆管细胞癌的发生。     

泡膜蛋白-1在星形细胞肿瘤中的表达以及与预后关系的研究

目的 探讨泡膜蛋白-1(vacuole membrane protein-1,VMP1)在星形细胞肿瘤中的表达,以及与星形细胞肿瘤患者预后的关系.方法 回顾分析经病理证实星形细胞肿瘤274例的资料,采用免疫组化法检测VMPI表达,随访患者生存预后情况.结果 274例中随访到212例,VMP1总阳性表达率为78.5%,不同级别星形细胞肿瘤VMP1表达阳性率随肿瘤恶性程度增加而降低,VMP1表达与生存时间呈正相关.结论 VMP1的表达强度对判断星形细胞肿瘤恶性程度和预后具有参考价值,可作为星形细胞肿瘤早期诊断、临床分级和顸后判断的一个重要生物学指标.     

Caveolin-1和E-cadherin在肾细胞癌中的表达及其意义

目的:探讨Caveolin-1、E-cadherin在肾细胞癌中的表达及其临床意义.方法:采用免疫组织化学EnVision法检测41例肾细胞癌切除标本中Caveolin-1和E-cadherin的表达,并分析表达情况与肿瘤的生物学行为的关系.结果:Caveolin-1在肾细胞癌中的表达阳性率为41.5%,与肿瘤分期、Fuhrman分级及患者年龄呈正相关(P<0.05);E-cadherin表达阳性率为24.4%,低于癌旁肾组织(50%),E-cadherin表达与肿瘤细胞核分级相关(P<0.05),与其他临床病理参数之间无相关性.Caveolin-1和E-cadherin两者之间表达无相关性.结论:Caveolin-1过表达及E-cadherin的异常表达,可能在肾细胞癌的生长、分化中发挥重要作用.     

细胞间粘附分子—1在膀胱移行细胞癌中的表达及意义

目的:探讨细胞间粘附分子-1(ICAM-1)在膀胱移行细胞癌中的表达及意义.方法:应用免疫组织化学方法对59例膀胱移行细胞癌中细胞间粘附分子-1的表达进行研究.结果:59例膀胱移行细胞癌中25例呈阳性表达,而正常膀胱粘膜未见表达,ICAM-1阳性表达的组织中常伴有大量淋巴细胞浸润.ICAM-1在膀胱移行细胞癌中的阳性表达率为42.37%,其阳性率在肿瘤病理分级、临床分期等方面差异有显著性(P<0.01).结论:ICAM-1在膀胱肿瘤中的表达与机体抗肿瘤免疫有关,同时可作为判断肿瘤恶性程度的指标.     

肺癌组织分型与转化生长因子-βⅡ型受体和肿瘤相关性巨噬细胞计数的关系

目的 检测肺腺癌和肺鳞状细胞癌中转化生长因子-βⅡ型受体(TGF-βRⅡ)和肿瘤相关性巨噬细胞(TAMs)计数,探讨两者与肺腺癌和肺鳞状细胞癌的关系.方法 应用免疫组织化学法检测TGF-βRⅡ蛋白和TAMs在肺腺癌和肺鳞状细胞癌组织中的表达,并与癌旁正常肺组织中的表达进行比较.结果 (1)TGF-βRⅡ蛋白的表达在肿瘤组织中明显降低(P＜0.01),其表达下调与肺癌的组织学类型、淋巴结转移、临床分期相关(P＜0.05).(2)TAMs在肿瘤间质内浸润数量明显增多(P＜0.01),并与肺癌的组织学类型、肿瘤大小、淋巴结转移、临床分期密切相关(P＜0.05).结论 TGF-βRⅡ蛋白表达水平降低可促进肺癌的发生、浸润和转移,对肺腺癌发生或进展影响很大,并可能通过下调TGF-βRⅡ的表达、诱导TAMs至肿瘤间质,从而促进肿瘤的发生、发展,且这种作用在非小细胞肺癌中具有较普遍的意义.     

MiR-34b在非小细胞肺癌中的表达及其在HGF-Met信号通路中的作用

目的 microRNAs (miRNAs)是内源性非编码小RNA,肿瘤细胞中大多数miRNAs表达降低,探讨miRNAs抑制肿瘤的机制.方法 应用实时定量PCR检测非小细胞肺癌组织miR-34b的表达,并且分析其与临床病理参数之间的关系.应用细胞转染技术获得miR-34b高表达细胞,并检测细胞凋亡与细胞增殖.运用免疫组织化学方法探索miR-34b表达与下游蛋白c-Met、p53以及Mdm2之间的相关性.结果 miR-34b在非小细胞肺癌中低表达,并与淋巴结转移负相关(P=O.031).miR-34b通过诱导凋亡抑制肿瘤增殖.非小细胞肺癌中miR-34b表达与c-Met负相关(P=0.012).结论 HGF-Met信号通路中p53磷酸化并增强转录活性,上调miR-34b,反馈抑制c-Met;miR-34b影响肿瘤细胞凋亡、增殖、转移,在肿瘤发展过程中发挥抑制肿瘤的作用.     

Secretagogin在神经内分泌肿瘤组织的表达

目的:通过检测Secretagogin(SCGN)在神经内分泌肿瘤组织中的表达,与神经内分泌肿瘤标志物进行比较,探讨SCGN作为临床病理诊断神经内分泌肿瘤标志物的可能性.方法:收集神经内分泌肿瘤181例(包括类癌、小细胞肺癌、肾上腺皮脂腺瘤等共14种)和非内分泌肿瘤198例(包括肺肿瘤共39种).所有标本分别用SCGN、CD56、Svn、CgA和NSE抗体进行免疫组织化学染色.结果:181例神经内分泌肿瘤中115例SCGN表达阳性,而且在类癌、小细胞肺癌、大细胞神经内分泌癌、癌伴神经内分泌分化、胰腺内分泌肿瘤、垂体腺瘤和甲状腺髓样癌中均呈持续阳性表达,阳性率均高于80%,明显高于CD56、CgA、SYN、NSE.而SCGN在非神经内分泌肿瘤中表达仅为3%.结论:SCGN对神经内分泌肿瘤具有高度特异性和敏感性,可能成为神经内分泌肿瘤标志物.     

蛋白酪氨酸磷酸酶-3通过诱导肿瘤性巨噬细胞钙离子依赖性钾离子通道表达增强LoVo细胞侵袭性

目的 观察结肠癌肿瘤微环境中肿瘤细胞与巨噬细胞(M2)相互作用及对肿瘤细胞功能学的改变.方法 将转入蛋白酪氨酸磷酸酶-3(PRL-3)的LoVo细胞和M2细胞模拟肿瘤微环境进行共培养,通过Western blot检测M2细胞钙离子依赖性钾离子通道(KCNN4)蛋白表达,并检测LoVo细胞侵袭性的改变.结果 Western blot检测示PRL-3能通过共培养后诱导M2细胞KCNN4蛋白表达升高,而未作共培养的M2细胞KCNN4蛋白表达未升高.此外,经过共培养后LoVo细胞侵袭性升高(3367±135比1442±89,P＜0.05),而在阻断KCNN4蛋白表达后,LoVo细胞侵袭性降低(1388 ±87比2893±163,P＜0.05).结论 PRL-3在结肠癌肿瘤微环境中通过提高KCNN4表达从而增强LoVo细胞的侵袭性.     

Fibulin-1基因表达与肾细胞癌的关系研究

目的 检测肾细胞癌组织中Fibulin-1基因表达的改变,探讨其相关的临床意义.方法 选取2014年1月至2016年5月本院收治的肾细胞癌患者手术切除种癌组织83例,以及相应的癌旁正常非瘤肾脏组织.SYBR Green 荧光定量-PCR检测癌组织和非癌组织Fibulin-1基因表达量.结果 肾细胞癌肿瘤组织中Fibulin-1基因表达量为(0.176±0.028),显著低于对照非肿瘤肾组织的(0.384±0.052,P<0.01).肾细胞癌肿瘤组织中Fibulin-1基因表达量在不同性别、不同年龄和组织病理类型间差异无统计学意义(P>0.05).有淋巴结转移肿瘤组织Fibulin-1基因表达量为(0.152±0.022),显著低于无淋巴结转移肿瘤组织(0.204±0.035,P<0.05).中、低分化肿瘤组织Fibulin-1基因表达量为(0.160±0.021),显著低于高分化肿瘤组织(0.207±0.036,P<0.05).Ⅲ、Ⅳ期肿瘤组织中的Fibulin-1基因表达量为(0.148±0.019),显著低于Ⅰ、Ⅱ期肿瘤组织(0.199±0.034,P<0.05).结论 肾细胞癌患者fibulin-1基因表达下调,fibulin-1基因表达下调与肾细胞癌有无淋巴结转移、分化程度以及临床分期密切相关.     

前列腺干细胞抗原在膀胱癌治疗中的研究进展

前列腺干细胞抗原(PSCA)为一种细胞表面抗原,除在正常前列腺中表达外,在前列腺癌、膀胱癌及部分消化道肿瘤表达,与肿瘤的发生、发展有重要关系.本文就前列腺干细胞抗原(PSCA)在膀胱肿瘤治疗中的研究进展进行综述.     

RON,a Tyrosine Kinase Receptor Involved in Tumor Progression and Metastasis

Tyrosine kinase receptors mediate many critical cellular functions that contribute to tumor progression and metastasis and thus are potential targets for molecular-based cancer therapy. As has been found for many receptor tyrosine kinases, RON (recepteur dorigine nantais) and its ligand, macrophage-stimulating protein, have recently been implicated in the progression and metastasis of tumors. In in vitro experiments using colon and breast cancer cell lines, overexpression of RON led to increased invasion and migration of cancer cells and prevented apoptosis and anoikis. In addition, transgenic mice engineered to overexpress RON in the lung epithelium developed multiple pulmonary tumors, suggesting a role for RON in tumorigenesis. In human cancer specimens, increased RON expression has been demonstrated in colon, breast, ovarian, and lung tumors. Therefore, therapies designed to inhibit RON activation may hinder critical tumor survival mechanisms and play a role in the treatment of advanced disease.Published by Springer Science+Business Media, Inc. © 2005 The Society of Surgical Oncology, Inc.     

Collaboration of RON and epidermal growth factor receptor in human bladder carcinogenesis

PURPOSE: Collaboration of heterologous receptor tyrosine kinases has emerged as an important paradigm in tumor progression. We recently proved that RON has an important role in human bladder carcinogenesis. Since epidermal growth factor receptor has been suggested to cross-talk with RON, we examined the significance of epidermal growth factor receptor in modulating RON associated tumorigenesis. MATERIALS AND METHODS: The biological significance of collaboration between RON and epidermal growth factor receptor was examined in the TSGH8301, J82 and JR bladder cancer cell lines with different expression status. Immunoprecipitation and immunoblotting assays were done to investigate the interaction of RON with epidermal growth factor receptor in relation to epidermal growth factor receptor kinase inhibitor treatment. Time lapse wound healing monitoring and Transwelltrade mark assay were used for cell migration analysis and the effect on cell transformation was analyzed with foci formation assay. Finally, a bladder cancer cohort of 78 patients was studied for clinical significance by immunohistochemistry. RESULTS: Epidermal growth factor receptor was directly associated with RON, irrespective of ligand stimulation. The siRNA experiment and epidermal growth factor receptor kinase inhibitors efficiently inhibited RON related biological effects, including mitogenesis, migration, anti-apoptosis and neoplastic transformation. Co-expression of RON/epidermal growth factor receptor was found in 26 of 78 patients (33.3%) with bladder cancer. It was significantly associated with tumor invasion (p < 0.05), the risk of local recurrence (p = 0.0003) and decreased patient survival (p = 0.04). Important indicators for patient survival were co-expression of RON and epidermal growth factor receptor (p = 0.001) and tumor staging (p = 0.05). CONCLUSIONS: Cross-talk between epidermal growth factor receptor and RON exists in vivo. Thus, it should be considered in treatment planning for patients with bladder cancer.     

Prognostic role of the recepteur d'origine nantais (RON) expression in primary high-grade osteosarcoma

BackgroundOsteosarcoma is a common primary malignant bone tumor susceptible to distant metastasis. The clinical outcome for patients remains poor due to the resistance to chemotherapy and lacking effective therapeutic targets. Recepteur d'origine nantais (RON), a transmembrane protein of the c-MET proto-oncogene family, has been reported to contribute to the malignant progression and bone metastasis in several tumors. The present study aimed to explore the prognostic significance of RON in primary high-grade osteosarcoma.MethodsImmunohistochemistry (IHC) and western blotting (WB) were used to investigate the protein expression of RON in 80 surgically resected specimens (50 high-grade osteosarcoma specimens and 30 non-neoplastic bone tissues) and 6 cell lines. The χ² test or independent-sample Student's t-test was used to assess the significance of RON difference between osteosarcoma and non-neoplastic bone tissues. The χ² test and Fisher's exact test were used to analyze the association of RON with the clinicopathological features of osteosarcoma patients. Kaplan–Meier method and Cox proportional hazards model were used to assess the significance of RON for the survival of osteosarcoma patients.ResultsThe results of IHC and WB observed significant overexpression of RON in osteosarcoma specimens (P < 0.001) and osteosarcoma cell lines. Moreover, immunohistochemical high expression of RON was associated with a poor response to chemotherapy (P = 0.032) as well as worse progression-free (P = 0.003) and overall (P < 0.001) survival of osteosarcoma patients. Multivariate analysis revealed that high expression of RON was independently associated with reduced progression-free (P = 0.027, HR = 2.31) and overall survival (P = 0.004, HR = 5.06) time of osteosarcoma patients.ConclusionsThe present study demonstrated that high expression of RON held independent value for unfavorable survival in primary high-grade osteosarcoma. Its potential role as a therapeutic target for osteosarcoma treatment deserves further research.     

表没食子儿茶素没食子酸酯抑制胃癌生长和血管生成及其分子机制的研究

目的探讨表没食子儿茶素没食子酸酯（EGCG）抑制胃癌生长和血管生成的分子机制。方法建立裸鼠异位胃癌肿瘤模型．检测肿瘤生长及肿瘤组织微血管密度（MVD）。培养SGC-7901胃癌细胞．Western印迹方法检测肿瘤细胞及其组织血管内皮生长因子（VEGF）、总信号转导、转录活化因子（Star3）和磷酸化形式Stat3的表达,同时采用ELISA方法检测肿瘤细胞培养液中VEGF蛋白水平．RT-PCR方法检测胃癌细胞VEGFmRNA的表达水平。结果EGCG组肿瘤组织平均质量（0．32±0．08）g,显著低于对照组（0．81±0．12）g,t=7．24,P〈0．01。EGCG组平均肿瘤抑制率为（60．4±6．1）％：其肿瘤生长曲线也显著低于对照组、EGCG组的肿瘤组织MVD（15．2±4．3）也显著低于对照组（24．6±6．6）（t=3．41,P〈0．01）。EGCG组胃癌组织的VEGF表达也减少78．6％．并呈剂量依赖性地下降：其肿瘤组织中Stat3磷酸化活化也减少了53．5％,也呈剂量依赖性地下降：但并未影响总的Stat3表达。结论EGCG通过抑制Stat3活化减少胃癌细胞VEGF表达．从而抑制胃癌生长和血管生成。     

Neutralization of macrophage-stimulating protein ameliorates renal injury in anti-thy 1 glomerulonephritis

Macrophage-stimulating protein (MSP) is a scatter factor that causes cell proliferation and migration, and receptor origin nantaise (RON) is its receptor. RON is expressed in macrophages and mesangial cells, and MSP is produced by renal tubular cells. This study investigated whether MSP/RON participate in the pathogenesis of anti-Thy 1 nephritis, a glomerular disease that is characterized by invasion of circulating monocytes into glomeruli and migration and proliferation of mesangial cells. In vivo, renal function and histopathology were studied in rats that had anti-Thy 1 disease and were untreated and treated with a neutralizing anti-MSP antibody. In vitro, whether monocytes express RON and whether MSP has a chemotactic effect on monocytes were studied. In vivo, in anti-Thy 1 disease, MSP was expressed de novo in glomeruli, and neutralization of MSP attenuated the rise in serum creatinine and proteinuria, stopped glomerular neutrophil and monocyte influx, protected from glomerular injury, and lessened mesangial cell overgrowth. In vitro, unstimulated monocytes did not express RON, but the stimulation with LPS induced de novo RON expression. LPS-stimulated monocytes were attracted by MSP. These results demonstrate a pathogenic role of the MSP/RON system in anti-Thy 1 nephritis.     

Soluble factors involved in glioma invasion

Summary ¶Recent studies using molecular and cellular techniques of the factors regulating the invasion process have revealed a crucial role for a number of growth factors and cytokines. Their function lies on the one hand in the autocrine stimulation of the tumor cells themselves, resulting in the stimulation of protease expression and an enhancement of migratory potential. On the other hand, the growth factors and cytokines seem to play a major role in the paracrine activation of the tumor surrounding stroma. Through stimulation of the strong angiogenic response that is characteristic for gliomas and also of the expression of proteases in the stromal cells, they contribute critically to the generation of a stromal environment that is permissive or even inductive for tumor cell invasion. Understanding of the mechanisms by which soluble factors modulate glioma cell invasion therefore will help to determine targets for the modification of existing therapies and lead to the development of novel therapeutic strategies in the management of gliomas.Published online November 3, 2003