Aberrant expression of the tumour suppressor gene p53 is very frequent in Wilms' tumours.

A series of 34 Wilms' tumours have been analysed for abnormal expression of the tumour suppressor gene p53 using frozen section immunohistochemistry. All tumours showed immunoreactivity with at least one of the specific antibodies used (monoclonal antibody PAb240, polyclonal antibodies CM1 and JG8). Abnormalities of p53 expression are very frequent in this type of childhood tumour.     

A new precursor lesion of Wilms' tumour (nephroblastoma): intralobar multifocal nephroblastomatosis

Almost half of the kidneys containing Wilms' tumours also contain persistent renal blastema in one or more of its forms. Persistent renal blastema is present in almost all cases of bilateral nephroblastoma. Multifocal superficial nephroblastemomatosis is the best known form of persistent renal blastema, and there is good evidence that it is a precursor of nephroblastoma. This paper reports four cases of a second, deep cortical type of nephroblastomatosis. This type of persistent renal blastema appears to be a precursor of a distinct histologic subtype of nephroblastoma, and differs from multifocal superficial nephroblastomatosis also in its teratological background.     

Expression and localization of HGF and met in Wilms' tumours.

A number of growth factors and cognate receptors that contribute to normal kidney development have been shown to play roles in the pathogenesis of Wilms' tumours. Expression of both hepatocyte growth factor (HGF) and its tyrosine kinase receptor met has been demonstrated in normal tissues and their neoplastic counterparts, implicating these factors in normal development and tumour progression. HGF and met expression has not been studied in Wilms' tumour. Since HGF and met function in a paracrine fashion by regulating tubulogenesis in normal kidney development, they could be involved in the pathogenesis of Wilms' tumour, in which tubular formation is dysplastic. In the present study, a series of ten homotypic (consisting of blastemal, epithelial, and stromal elements) and ten heterotypic (consisting of triphasic histology and a muscle component) Wilms' tumour cases were examined for expression of HGF and met, using in situ hybridization, immunohistochemistry, and western blot analysis. Relatively high met message and protein expression, compared with normal kidney, were evident in homotypic and heterotypic tumour blastemal, epithelial, and rhabdomyoblastic cells and a 145 kD met polypeptide was found in all tumours, with a few cases also expressing the 170 kD precursor form. No apparent alterations of the met receptor were observed. Similarly, HGF protein was also abundantly expressed in blastemal, epithelial, and rhabdomyoblastic cells of the homotypic and heterotypic Wilms' tumours and a 69 kD HGF polypeptide was demonstrated by western blot analysis. Immunohistochemistry for the Ki-67 proliferation marker indicated that the pattern of Ki-67 expression correlated with the HGF and met pattern of expression in both homotypic and heterotypic tumours. These results reveal, for the first time, significant co-expression of met/HGF in Wilms' tumours, with a correspondingly high proliferative index in the same cell groups.     

Simultaneous blockade of programmed death 1 and vascular endothelial growth factor receptor 2 (VEGFR2) induces synergistic anti‐tumour effect in vivo

Recent basic and clinical studies have shown that the programmed death ligand (PD‐L)/PD‐1 pathway has a significant role in tumour immunity, and its blockade has a therapeutic potential against several human cancers. We hypothesized that anti‐angiogeneic treatment might augment the efficacy of PD‐1 blockade. To this end, we evaluated combining the blockade of PD‐1 and vascular endothelial growth factor receptor 2 ( VEGFR2) in a murine cancer model using Colon‐26 adenocarcinoma. Interestingly, simultaneous treatment with anti‐PD‐1 and anti‐VEGFR2 monoclonal antibodies (mAbs) inhibited tumour growth synergistically in vivo without overt toxicity. Blocking VEGFR2 inhibited tumour neovascularization significantly, as demonstrated by the reduced number of microvessels, while PD‐1 blockade had no impact on tumour angiogenesis. PD‐1 blockade might promote T cell infiltration into tumours and significantly enhanced local immune activation, as shown by the up‐regulation of several proinflammatory cytokine expressions. Importantly, VEGFR2 blockade did not interfere with T cell infiltration and immunological activation induced by PD‐1 blockade. In conclusion, simultaneous blockade of PD‐1 and VEGFR2 induced a synergistic in‐vivo anti‐tumour effect, possibly through different mechanisms that might not be mutually exclusive. This unique therapeutic strategy may hold significant promise for future clinical application.     

Analysis by array CGH of genomic changes associated with the progression or relapse of Wilms' tumour

Despite aggressive salvage regimens, approximately half of all children who suffer a Wilms' tumour recurrence will die of their disease. Although there are increasing data on molecular genetic prognostic factors present in the tumour at diagnosis, there is little information regarding the molecular events that occur with Wilms' tumour progression and relapse. In the present study, microarray-based comparative genomic hybridization (aCGH) analysis has been carried out on 58 Wilms' tumour samples, which included 38 untreated primary and 20 recurrent tumours. A higher degree of copy number changes was observed in the recurrent tumours (33.0% genomic clones) than in the primary tumour (21.2%). Paired analysis highlighted the acquisition of 15q gain with high levels of IGF1R expression in the tumour recurrence in two cases. The most statistically significant abnormality acquired between diagnosis and relapse was loss of 17p. One case that experienced 17p loss was classified as favourable histology at diagnosis, but exhibited diffuse anaplasia at recurrence and had a homozygous TP53 deletion. Another instructive case with a constitutional 11p13 deletion presented with bilateral tumours and suffered two subsequent recurrences in the left kidney. A somatic WT1 mutation was found only in the right kidney tumour, while the constitutional 11p13 deletion was the only abnormality detected in the initial left kidney tumour by aCGH. The two subsequent relapses in the left kidney contained an accumulation of additional genetic alterations, including an independent WT1 mutation.     

VEGF‐C,VEGF‐D and VEGFR‐3 expression in peripheral neuroblastic tumours

Ramani P, Nash R, Radevsky L, Patel A, Luckett M & Rogers C
(2012) Histopathology
VEGF‐C, VEGF‐D and VEGFR‐3 expression in peripheral neuroblastic tumours Aims: More than 50% of neuroblastomas (NBs) present with haematogenous and/or lymphatic metastasis; however, little is known about the clinicopathological significance in NBs of the key lymphangiogenesis growth factors vascular endothelial growth factor (VEGF)‐C and VEGF‐D and the receptor VEGFR‐3. Methods and results: Ninety‐three NBs and nine ganglioneuromas (GNs) were immunostained for VEGF‐C, VEGF‐D and VEGFR‐3. VEGF‐C and VEGF‐D were present in 76% and 82% of the NBs, respectively. There was no significant difference in VEGF‐C expression between NBs and GNs. VEGF‐D expression was significantly higher in NBs compared with GNs and in MYCN‐amplified NBs. VEGFR‐3 tumoral cell expression (VEGFR‐3c), present in 48% of the NBs, was significantly higher in NBs from children ≥18 months at presentation and those belonging to a high‐risk group. VEGFR‐3 lymphovascular density was increased significantly in NBs compared with GNs and in NBs associated with adverse clinicopathological and biological factors. Lymphovascular invasion, assessed in VEGFR‐3‐stained vessels, was present in ～50% of NBs. Cox regression analyses demonstrated that VEGFR‐3c expression was associated with a significantly shorter event‐free survival and that its effect was independent of the important pathological variable, mitosis–karyorrhexis index. Conclusions: VEGF‐D and VEGFR‐3 up‐regulation support tumour progression in NB and VEGFR‐3c may provide a useful prognostic marker in NBs.     

VEGF与MMP-2在食管鳞癌中的表达及其临床病理意义

目的研究血管内皮生长因子（VEGF）和基质金属蛋白酶-2（MMP-2）在食管鳞癌中的表达与其临床意义。方法用免疫组化染色的方法检测58例食管鳞癌标本及30例取非瘤正常食管组织标本中VEGF与MMP-2蛋白的表达。结果食管鳞癌中VEGF与MMP-2的表达显著高于正常组织，其表达与食管癌的组织侵润深度、淋巴结转移密切相关，两者呈正相关。结论VEGF和MMP-2在食管鳞癌中高表达，参与了食管鳞癌浸润及转移，可以作为判断食管鳞癌生物学行为的指标之一。     

The prognostic significance of angiogenesis and the effect of vascular endothelial growth factor on angiogenic process in Wilms' tumour

AIMS: There is a subgroup of patients with Wilms' tumour (WT) having favourable clinicopathological features but adverse outcome. We aimed to investigate the prognostic significance of angiogenesis and whether it can be used for predicting which patients will fall into this category, and the possible role of vascular endothelial growth factor (VEGF) on angiogenesis in WT. METHODS: Tumours in nephrectomy specimens from 63 WT patients were investigated for neovascularisation and VEGF expression by immunohistochemistry. The endothelial cells were highlighted by anti-CD34 and anti-CD31, and the microvessels in the hot-spots were counted. Correlations between the microvessel density (MVD), VEGF expression, clinicopathological features and prognosis were studied. RESULTS: Among 21 patients with follow-up data, favourable histology was detected in 17, seven of which died of disease. Patients with highly vascular tumours showed significantly poorer prognosis than those with low vascular tumours. There was no significant relationship between angiogenesis and VEGF expression. VEGF immunostaining revealed various patterns in different components of WT. CONCLUSIONS: We suggest that high MVD can be used as an indicator of poor prognosis with WT patients displaying favourable histology and there might be some additional growth factors other than VEGF which may also be responsible for angiogenesis in WTs.     

The prognostic significance of growth factors and growth factor receptors in gastric adenocarcinoma

We evaluated growth factors/receptors expression in gastric adenocarcinoma. Immunohistochemistry was used to evaluate epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), VEGF‐D, VEGF receptor (VEGFR)‐2, VEGFR‐3, transforming growth factor (TGF)‐α, TGF‐β1, and TGF‐β‐RII in tissue microarrays of adenocarcinoma, dysplasia, metaplasia, and gastritis. In adenocarcinoma, the expression rates of EGFR, VEGF, VEGF‐D, VEGFR‐2, VEGFR‐3, TGF‐α, TGF‐β1, and TGF‐β‐RII were 2.0%, 0%, 10.7%, 4.4%, 11.2%, 26.3%, 9.4%, and 19.5%, respectively. VEGF‐D, TGF‐α, TGF‐β1, and TGF‐β‐RII expression rate were higher in adenocarcinoma than in other groups. TGF‐β‐RII expression was correlated with VEGFR‐3, VEGF‐D, and TGF‐α expression in adenocarcinomas. Tumor location, histologic type, stage, lymphatic invasion, perineural invasion, angioinvasion, VEGF‐D, and VEGFR‐2 expressions were associated with patient survival in a log rank test and advanced stage and positive expression of VEGF‐D were poor prognostic factors using Cox analysis. VEGF‐D expression may be of prognostic value in gastric adenocarcinoma, whereas EGFR and TGF family expression may only have a minor influence.     

COX-2、VEGF在乳腺癌组织中的表达及临床意义

目的探讨血管内皮生长因子(VEGF)和环氧化酶-2(COX-2)在乳腺癌组织中的表达及其与临床病理特征之间的关系。方法应用免疫组化S-P法检测不同乳腺组织中VEGF和COX-2的表达情况。结果VEGF、COX-2在乳腺导管癌组织中的表达明显上调,与其在正常乳腺组织、纤维腺瘤组织中的表达相比差异显著(P<0.01)。VEGF、COX-2在乳腺浸润性导管癌中的阳性表达率高于其在乳腺导管癌中的阳性表达率。两组中COX-2的表达有显著性差异(77.1%vs40%,P<0.05),VEGF的表达无显著性差异(80.0%vs70%,P>0.05)。VEGF在乳腺浸润性导管癌的阳性表达与其临床分期、淋巴结转移密切相关(P<0.05,P<0.01),与肿瘤大小无关(P>0.05)。COX-2在乳腺浸润性导管癌中的阳性表达与其临床分期、淋巴结转移及肿瘤大小均无关(P>0.05)。VEGF、COX-2在乳腺导管癌中的表达呈正相关(r=0.98,P<0.05)。结论VEGF、COX-2的高表达在乳腺癌的发生发展过程中起重要的作用,可作为重要的生物学标志。     

Expression pattern of VEGFR-2 (Quek1) during quail development

The growth and maintenance of the blood and lymphatic vascular systems is to a large extent controlled by members of the vascular endothelial growth factor (VEGF) family via the tyrosine kinase receptors (VEGFRs) expressed in angioblastic cells. Here, we analyzed the Quek1 (VEGFR-2) expression pattern by whole mount in situ hybridization during quail development. During early embryogenesis, Quek1 expression was detected at the caudal end of the blastoderm and primitive streak and in the head paraxial mesoderm. In somites, expression was observed from HH-stage 9 onwards in the dorsolateral region of both the forming and recently formed somites. During somite maturation, expression persists in the lateral portion of the somitic compartments, the dermomyotome and the sclerotome. Additionally, a second expression domain in the maturing somite was observed in the medial part of the sclerotome adjacent to the neural tube. This expression domain extended medially and dorsally and surrounded the neural tube during later stages. In the notochord, expression was observed from HH-stage 23 onwards. In the limb bud, expression was initiated in the mesenchyme at HH-stage 17. During organogenesis, expression was detected in the pharyngeal arches and in the anlagen of the esophagus, trachea, stomach, lungs, liver, heart and gut. Expression was also seen in feather buds from day 7 onwards. Our results confirm the angiogenic potential of the mesoderm and suggest that VEGFR-2 expressing cells represent multiple pools of mesodermal precursors of the hematopoietic and angiopoietic lineages.     

Circulating tumour tissue fragments in patients with pulmonary metastasis of clear cell renal cell carcinoma

Tumour metastasis is the result of a complex sequence of events, including migration of tumour cells through stroma, proteolytic degradation of stromal and vessel wall elements, intravasation, transport through the circulation, extravasation and outgrowth at compatible sites in the body (the ‘seed and soil’ hypothesis). However, the high incidence of metastasis from various tumour types in liver and lung may be explained by a stochastic process as well, based on the anatomical relationship of the primary tumour with the circulation and mechanical entrapment of metastatic tumour cells in capillary beds. We previously reported that constitutive VEGF‐A expression in tumour xenografts facilitates this type of metastatic seeding by promoting shedding of multicellular tumour tissue fragments, surrounded by vessel wall elements, into the circulation. After transport through the vena cava, such fragments may be trapped in pulmonary arteries, allowing them to expand to symptomatic lesions. Here we tested whether this process has clinical relevance for clear cell renal cell carcinoma (ccRCC), a prototype tumour in the sense of high constitutive VEGF‐A expression. To this end we collected and analysed outflow samples from the renal vein, directly after tumour nephrectomy, in 42 patients diagnosed with ccRCC. Tumour fragments in venous outflow were observed in 33% of ccRCC patients and correlated with the synchronous presence or metachronous development of pulmonary metastases (p < 0.001, Fisher's exact test). In patients with tumours that, in retrospect, were not of the VEGF‐A‐expressing clear cell type, tumour fragments were never observed in the renal outflow. These data suggest that, in ccRCC, a VEGF‐A‐induced phenotype promotes a release of tumour cell clusters into the circulation that may contribute to pulmonary metastasis. Copyright © 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

血管内皮生长因子-C和血管内皮生长因子受体-3在侵袭性甲状腺乳头状癌中的表达及意义

目的 观察血管内皮生长因子(VEGF-C)和血管内皮生长因子受体(VEGFR3)在侵袭性甲状腺乳头状癌组织中的表达,探讨甲状腺乳头状癌细胞淋巴管转移机理.方法 用免疫组化方法检测VEGF-C和VEGFR3在人侵袭性甲状腺乳头状癌和非侵袭性甲状腺乳头状癌中的表达情况,并进行比较和分析.结果 在侵袭性和非侵袭性甲状腺乳头状癌中均可见到VEGF-C和VEGFR3阳性表达,但在侵袭性甲状腺乳头状癌组织VEGF-C (x2=4.738,P=0.030)和VEGFR3(x2=11.951,P=0.010)的表达率和表达强度均高于非侵袭性甲状腺乳头状癌.结论 VEGF-C和VEGFR3在侵袭性甲状腺乳头状癌细胞中高表达,并且可能与此肿瘤的侵袭性相关.     

Specificity of DOG1 (K9 clone) and protein kinase C theta (clone 27) as immunohistochemical markers of gastrointestinal stromal tumour

Wong N A C S & Shelley‐Fraser G
(2010) Histopathology 57 , 250–258 Specificity of DOG1 (K9 clone) and protein kinase C theta (clone 27) as immunohistochemical markers of gastrointestinal stromal tumour Aims: DOG1 and protein kinase C (PKC) theta are both sensitive immunohistochemical markers of gastrointestinal stromal tumour (GIST). However, there are conflicting data regarding the specificity of the most commonly used PKC theta antibody (clone 27), and there are no existing data regarding the specificity of the only known commercially available DOG1 antibody (K9 clone) at the time of writing. This study’s aim was to characterize the immunoreactivity patterns of both monoclonal antibodies amongst a wide range of neoplasm types including, in particular, histological mimics of GIST. Methods and results: Immunohistochemistry for DOG1 and PKC theta was performed on whole tissue sections from 23 different neoplasm types (total of 125 cases). Ten of these neoplasm types showed CD117 immunopositivity. Only three (Ewing’s sarcoma, glomus tumour and synovial sarcoma) of the 23 neoplasm types showed DOG1 immunopositivity, and such positivity was often focal and weak in intensity. In contrast, all but four (ganglioneuromas, leiomyomas, desmoplastic small round cell tumours and PEComa/angiomyolipomas) of the 23 neoplasm types showed PKC theta immunopositivity. Conclusions: Compared with CD117, DOG1 (using the K9 antibody) is a more specific marker, whereas PKC theta (using the clone 27 antibody) is a considerably less specific immunohistochemical marker for GIST.     

VEGF-C、VEGFR-3和iNOS在人乳腺癌淋巴管的表达

目的检测乳腺癌组织血管内皮生长因子-C(VEGF-C)及其受体(VEGFR-3)、iNOS基因表达的相关性及这三个基因mRNA表达水平与淋巴管密度(LVD)的相关性,为阐明乳腺癌淋巴管生成的分子机理提供实验依据。方法RT-PCR检测乳腺癌组织及正常乳腺组织VEGF-C、VEGFR-3、iNOSmRNA的表达水平;免疫组织化学染色法检测淋巴管内皮细胞上VEGFR-3的表达,测定淋巴管密度。结果乳腺癌LVD为20.35±4.23,显著高于正常对照组(P<0.05);乳腺癌组织VEGF-C、VEGFR-3、iNOSmRNA三者的表达率分别为74.0%、84.0%、82.0%,显著高于正常对照组(P<0.05);VEGF-C、VEGFR-3、iNOS阳性组中LVD分别为21.34±3.45、18.54±4.68、17.43±4.76,显著高于对照组(P<0.05)。结论VEGF-C、VEGFR-3、iNOSmRNA表达与淋巴管密度之间呈正相关,并且3者之间的表达亦具有相关性,这些基因的表达增高可能在乳腺癌淋巴管生成过程中具有重要作用。