Correlation between tumor suppressor inhibitor of growth family member 4 expression and microvessel density in breast cancer

Suppression of cell spreading and migration by inhibitor of growth 4 suggests that its loss may induce metastasis. Inhibitor of growth 4 expression level in 60 breast cancer tissues, 30 normal adjacent tissues, and tissues from patients with benign hyperplasia was determined by real-time polymerase chain reaction, Western blot, and immunohistochemistry. The correlation between inhibitor of growth 4 expression and clinical stage, histologic grade, and microvessel density in breast cancer was analyzed. Inhibitor of growth 4 messenger RNA and protein expression in breast cancer was significantly lower than that observed in adjacent normal and hyperplastic breast tissues (P < .05). Inhibitor of growth 4 expression decreased with increasing clinical stage and histologic grade. Moreover, the presence of lymph node metastasis was correlated with decreased inhibitor of growth 4 messenger RNA expression (P < .01), and a negative correlation was noted between inhibitor of growth 4 protein expression and microvessel density in breast cancer. Inhibitor of growth 4 may represent an important biomarker for assessing the severity of breast cancer. Further studies are required to fully evaluate the diagnostic and possible prognostic value of determining inhibitor of growth 4 levels in breast cancer.     

Increased expression of CD146 and microvessel density (MVD) in invasive micropapillary carcinoma of the breast: Comparative study with invasive ductal carcinoma-not otherwise specified

Invasive micropapillary carcinoma (IMPC) is a rare variant of ductal carcinoma of the breast, and is characterized by a high metastatic potential and an aggressive clinical course. Studies of CD146 expression and function in breast cancer remain scarce. The aim of this study was to evaluate the role of CD146 and microvessel density (MVD) in breast IMPC. CD146 mRNA expression and immunohistochemistry for CD146 and MVD measured by CD31 were assessed in 82 cases of IMPC and 137 cases of invasive ductal carcinoma, not otherwise specified (IDC-NOS). The mRNA level of CD146 in cancer specimens was higher in IMPC than in IDC-NOS. CD146 expression in tumor cells was up-regulated in IMPC as compared with that in IDC-NOS, and was positively correlated with histological grade, ER, PR status, and P53 expression in IMPC and IDC-NOS. CD146 expression in vascular endothelial cells was significantly higher than that in IDC, and was positively correlated with tumor progression in IMPC and IDC-NOS. MVD in IMPC was significantly higher than that in IDC. CD146 expression in tumor cells was positively correlated with that in vascular endothelial cells of IMPC and IDC-NOS. The association of CD146 expression with MVD and its correlation with progression in breast carcinoma indicated that CD146 is a potentially useful prognostic marker for breast cancer. CD146 could be a new drug target in the treatment of breast cancer.     

Musashi 1在结肠癌中的表达及其与微血管密度及预后的关系

目的:探讨Musashi 1在结肠癌中的表达与肿瘤微血管密度(MVD)及预后的关系。方法:应用实时荧光定量PCR的方法检测36对结肠癌组织及其配对的正常结肠组织中Musashi 1 mRNA的表达情况。通过免疫组化法对96例结肠癌组织中Musashi 1表达情况和微血管密度(MVD)进行检测。结合随访术后生存情况,统计分析Musashi 1表达与结肠癌病理参数、预后及MVD之间的相关性。结果:实时荧光定量PCR法结果显示,在结肠癌组织中,Musashi 1 mRNA的相对表达水平显著高于正常结肠组织(P0.05)。免疫组化结果显示,96例结肠癌组织中,有61例(63.5%)高表达Musashi 1。Musashi1的表达与患者性别、肿瘤大小、分化程度无相关性(P0.05),而与肿瘤的浸润深度、Dukes分期、淋巴转移、TNM分期密切相关(P0.05)。Kaplan-Meier生存分析显示,Musashi 1高表达患者的5年生存率显著低于Musashi 1低表达者(23.0%vs60.0%;P0.01),且Musashi 1的表达与结肠癌血管密度相关(P0.01)。结论:结肠癌中Musashi 1的表达较正常癌旁组织高,具有统计学意义。结肠癌中Musashi 1的表达与肿瘤浸润深度、Dukes分期、淋巴转移、TNM分期和血管密度密切相关。Musashi 1蛋白可通过促进瘤内血管增生,从而促进结肠癌的生长和侵袭转移,进一步影响结肠癌患者的预后。     

血管内皮生长因子-D、 微血管密度及微淋巴管密度的表达与乳腺癌临床病理特征的相关性

目的 探讨乳腺癌中血管内皮生长因子D(vascular endothelial growth factor-D,VEGF-D)及微血管密度(microvessel density,MVD)、微淋巴管密度(Lymphatic vessel density,MLD)表达情况及意义.方法 收集2009年1月至2016年5月在临沂市肿瘤医院病理科存档病例,其中乳腺癌61例,乳腺增生症23例.采用免疫组化法检测各组织VEGF-D、MVD和MLD表达.结果 乳腺癌组织中VEGF-D蛋白阳性率为62.30%,明显高于乳腺增生组织,差异有统计学意义(χ2=16.215,P<0.05);乳腺癌组织中MVD和MLD分别为(17.70±7.10)和(2.41±0.85),明显高于乳腺增生组织,差异有统计学意义(t=10.900、t=8.795,P<0.05);VEGF-D表达与乳腺癌患者年龄、肿瘤大小以及淋巴结转移无明显关系(χ2=0.394、0.032、0.244,P>0.05);MVD和MLD密度与乳腺癌患者年龄和肿瘤大小无明显差异(χ2=0.081,0.126,0.219,0.196,P>0.05);VEGF-D、MVD和MLD密度与乳腺癌组织学分级相关(χ2=13.076、23.892、10.082,P<0.05),随着组织学分级递增,VEGF-D、MVD和MLD密度明显增高;MVD和MLD在乳腺癌伴淋巴结转移中密度明显高于无淋巴结转移(t=2.481、5.791,P<0.05).结论 VEGF-D在乳腺癌组织中呈高表达,可能在乳腺癌的发生发展中起重要作用,而MVD和MLD与乳腺癌肿瘤分化、转移有一定的关系.     

Differential expression of hypoxia and (lymph)angiogenesis-related genes at different metastatic sites in breast cancer

Introduction Breast cancer can metastasize via lymphatic and hematogenous pathways. Hypoxia and (lymph)angiogenesis are closely related processes that play a pivotal role in the tumor progression and metastasis. The aim of this study was to compare expression of hypoxia and (lymph)angiogenesis-related genes between primary breast tumors and metastases in different tissues. Materials and methods A gene list of 269 hypoxia and (lymph)angiogenesis-related genes was composed and validated using Onto-Express, Pathway-express and Ingenuity software. The expression of these genes was compared in microarray data of 62 samples of primary tumors and metastases of 31 patients with breast cancer retrieved from Gene Expression Omnibus. Similarity between samples was investigated using unsupervised hierarchical clustering analysis, principal component analysis and permutation testing. Differential gene expression between primary tumors and metastases and between metastases from different organs was analyzed using Kruskall–Wallis and Mann–Whitney statistics. Results Unsupervised hierarchical cluster analysis demonstrated that hypoxia and (lymph)angiogenesis-related gene expression was more similar between samples from the same patient, than between samples from the same organ. Principal component analysis indicated that 22.7% and 7.0% of the total variation in the gene list was respectively patient and organ related. When differences in gene expression were studied between different organs, liver metastases seemed to differ most from the other secondary sites. Some of the best characterized molecules differentially expressed were VEGFA, PDGFRB, FGF4, TIMP1, TGFB-R1 and collagen 18A1 (precursor of endostatin). To confirm the results of these experiments at the protein level, immunohistochemical experiments were performed with antibodies for VEGFA and MMP-2. Conclusions Our results suggest that hypoxia and (lymph)angiogenesis-related gene expression is more dependent on the characteristics of the primary tumor than on the characteristics of the organs that bear the metastasis. However, when different organs are compared, the expression in liver metastases differs most from other metastatic sites and primary tumors, possibly due to organ-specific angiogenic and lymphangiogenic responses to metastasis-related hypoxia. Electronic Supplementary Material The online version of this article (doi: ) contains supplementary material, which is available to authorized users.     

Prognostic significance of microvessel density and vascular endothelial growth factor expression in sinonasal carcinomas

The prognostic significance of microvessel density and proliferative activity of the neoplastic cells, evaluated respectively by CD31 and Ki-67 positivity, and immunohistochemical expression of vascular endothelial growth factor (VEGF) was retrospectively investigated in 105 cases of sinonasal carcinoma (80 surgical specimens and 25 biopsies). The most represented histologic types were intestinal-type adenocarcinoma found in 36 patients (34.3%), squamous cell carcinoma (SCC) in 34 (32.4%), mucinous adenocarcinoma (mainly made up of signet-ring cell patterns) in 15 (14.3%), and adenoid cystic carcinoma in 7 (6.7%). Microvessel density values (in vessels per square millimeter), VEGF, and Ki-67 were not dependent on histologic type but were rather correlated to the histologic grading in SCC. Clinical data were available for 92 (87.6%) of 105 patients, with minimum follow-up of 48 months. Most of the patients (81.5%) were at an advanced stage (T3-T4) at diagnosis. The values of all markers were correlated to tumor stage (P = .03). Multivariate analysis showed that both microvessel density and proliferative activity of the neoplastic cells were independent prognostic parameters (mortality hazard ratio, 1.33 and 1.60, respectively). Although VEGF expression was not correlated to prognosis on the whole series (P = .06), it was a powerful prognostic marker when the analysis was restricted to the group of SCCs (hazard ratio, 3.02; 90% confidence interval, 1.58-5.80). These results show that tumor neoangiogenesis, expressed by microvessel density, together with proliferative activity, is a pathologic marker with a strong prognostic impact in sinonasal carcinomas. Therefore, it may be a useful tool in this field so as to carry out therapeutic protocol planning, which may be further enhanced by the adoption of the more recent antiangiogenic molecules.     

A study into methodology and application of quantification of tumour vasculature in rectal cancer

The application of new surgical techniques in combination with preoperative radiotherapy has minimised the risk of local recurrence in rectal cancer. However, distant metastasis is still a serious problem after seemingly curative resection in patients with rectal cancer. The present study aimed to evaluate the methodology for quantification and the characteristics of the tumour vasculature in relation to the development of metastasis in patients with rectal cancer. From a large multicentre trial, 88 patients were selected, ensuring a relatively high percentage of metastasis. This selection facilitates the study of tumour vasculature characteristics in relation to metastasis. Vessel number, perimeter and area were assessed at both the invasive front and intratumoural area. Hot-spot and random selections were performed simultaneously. The median of each vessel parameter in the study population was used to separate patients into a low- and high-vessel group. Differences in development of distant metastasis were studied between low- and high-vessel groups. The data of the present study show that only vascular perimeter randomly assessed at the invasive front was associated with distant metastasis. Patients with a high score had a lower distant metastasis rate than patients with a low score (37% and 62%, respectively). High-vessel perimeter was independent of tumor node metastasis staging, but was associated with an increased presence of immune cells, comprising T cells, mast cells, eosinophils and neutrophils. This methodological study on the biological relevance of various vessel characteristics showed that a large vascular endothelial surface, as reflected by a high perimeter, was the only vessel characteristic indicative of improved patient outcome. The underlying principle for this association may be the improved immune response.     

脑星形细胞瘤MMP-9、Flt-1表达与微血管密度的关系及其意义

目的探讨不同组织学类型的脑星形细胞瘤MMP-9、Flt-1表达与微血管密度（MVD）和恶性程度的关系。方法采用免疫组化方法检测50例星形细胞瘤中MMP-9、Fit-1和CD34的表达,并通过CD34免疫组化结果测定肿瘤组织中MVD。结果MMP-9、Flt-1在各型脑星形细胞瘤中均有不同程度的表达;不同组织学类型,其阳性表达的强度存在着明显的差异。在WHO分级中Ⅲ-Ⅳ级脑星形细胞瘤MMPO的阳性表达为（76．52&#177;3．38）％,Fit-1为（72．90&#177;2．64）％,MVD为87．00&#177;4．18;Ⅰ-Ⅱ级MMP-9的阳性表达为（27．90&#177;2．53）％,Fit-1为（29．67&#177;2．87）％,MVD为48．62&#177;4．05。Ⅲ～Ⅳ级MMP-9、Flt-1和MVD的表达明显高于Ⅰ～Ⅱ级（P〈0．01）;三者的表达呈正相关。结论MMPO、Flt-1表达和MVD与脑星形细胞瘤的组织学类型及恶性程度相关,有可能做为判断肿瘤恶性程度及预后的辅助指标。     

Comparative evaluation of microvessel density determined by CD34 or CD105 in benign and malignant gastric lesions

Microvessel density (MVD) is regarded as a surrogate marker for angiogenesis and has been used for tumor prognosis. In this study, MVD was identified immunohistochemically by monoclonal antibodies against CD105 and CD34 in the tissues representing gastric carcinoma, chronic gastritis, and hyperplastic polyps, and the results were correlated with clinicopathologic features. The expression of CD105 in the microvessels within benign lesions was barely visible, and MVD was markedly lower than that determined by CD34. CD34 was strongly expressed in the microvessels within hyperplastic polyps and tissues with gastritis. In gastric carcinoma, CD105 expression in microvessels was as high as the MVD, compared with benign lesions. CD105 stained well-formed mature and newly formed immature vessels within the cancer mass. Correlation analysis showed that MVD determined by CD105 correlated with blood vessel invasion, distant metastasis, and formation of ascites. Survival analysis demonstrated an inverse correlation between MVD count and overall survival: patients with MVD counts of 32 or higher survived for a much shorter time than those with counts lower than 32. Multivariate analysis confirmed that MVD determined by CD105 was an independent prognostic factor for survival. Microvessel density determined by CD34 inversely correlated with overall survival, but it did not correlate with other clinicopathologic parameters except formation of ascites. In conclusion, CD34 was universally expressed in blood vessels within benign and malignant tissues, whereas CD105 expression was minimal in benign tissues but stronger in gastric carcinoma. These data suggest that both CD105 and CD34 could be used for quantification of angiogenesis, but preference should be given to CD105 in the evaluation of prognosis in gastric carcinoma.     

Expression and association of carbonic anhydrase IX and cyclooxygenase-2 in colorectal cancer

Background

This work was designed to determine the relationship between hypoxia-inducible protein carbonic anhydrase IX (CA IX) and pro-inflammatory enzyme cyclooxygenase-2 (COX-2) in patients with colorectal cancer (CRC).

DEC1 (STRA13) protein expression relates to hypoxia- inducible factor 1-alpha and carbonic anhydrase-9 overexpression in non-small cell lung cancer

Differentiated embryo-chondrocyte expressed gene 1 (DEC1) is involved in cell differentiation, proliferation, and apoptosis, and was recently shown to be regulated by hypoxia. The present immunohistochemical study demonstrates extensive nuclear expression of the protein in 38% of a series of 115 non-small cell lung carcinomas using a polyclonal antibody (Ab) recognizing DEC1 protein. Such expression was directly related to the expression of two hypoxia-regulated proteins, namely the hypoxia-inducible factor (HIF) 1alpha and carbonic anhydrase-9. Although DEC1 was not related to angiogenesis or to the expression of VEGF and thymidine phosphorylase, a direct association with up-regulated bFGF receptors was noted. DEC1 was persistently expressed in the nuclei of normal bronchial and alveolar tissue. It is suggested that loss of DEC1 expression is an early event in the development of lung cancer, while DEC1 gene expression occurs in a subset of tumours and parallels the overexpression of other hypoxia-regulated proteins.     

胰腺癌中肝细胞生长因子表达与微血管密度的临床研究

目的探讨在胰腺癌中肝细胞生长因子（HGF）阳性表达及微血管密度（MVD）与肿瘤临床病理特征的关系。方法应用免疫组化（S-P）法检测30例胰腺癌组织中HDF表达及MVD,运用统计软件将其与临床病理因素间进行Х^2及t检验。结果HGF不同分期下的表达不同,随分期增加而增加（P=0．04）;MVD在胰腺癌组织中随分期增加而增加（P=0．01）。肿瘤合并淋巴结转移患者HGF阳性表达为76．92％,与没有淋巴结转移组织间比较差异有统计学意义（P=0．001）;合并淋巴结转移组织内MVD计数为每高倍镜视野下（91．556±13．173）条,与没有淋巴结转移组织间比较差异有统计学意义（P=0．01）。结论HGF促进细胞的增生;促进肿瘤细胞的迁移,增加瘤细胞的侵袭性;还可通过诱导血管内皮生长因子的分泌和表达,从而间接促进肿瘤新生血管生成而促进肿瘤增长、转移。     

CD34 microvessel density and VEGF expression in basal and squamous cell carcinoma

Angiogenesis is a central process in the growth of solid tumors. The purpose of our study was to analyze the angiogenic pattern in squamous and basal cell carcinomas and to point out differences in microvessel density that could explain their different biological behaviour. Thirty-nine skin tumors (26 basal and 13 squamous cell carcinomas) were analyzed. In all samples, the microvessels density (MVD) and the levels of vascular endothelial growth factor mRNA (VEGFmRNA) were analyzed, together with the inter-relationship between these two variables. Using the median value of the entire series (33 vessels per 2.22 mm2), tumors with low and high MVD were identified. The majority of cancers with high vascularization belonged to the squamous histotype (12 of 39), while 19 of the 26 basal cell carcinomas showed a lower number of microvessels than the median value (p = 0.0001). The median value of VEGFcDNA quantitation allowed us to distinguish tumors with high VEGF expression (> 470 molecules cDNA) from those with low (< or = 470 molecules) VEGF expression: 20 of the 26 basal cell carcinomas showed low VEGF expression, while 11 of the 13 squamous cell carcinomas showed high VEGFcDNA levels (p = 0.0003). Moreover, a significant association between a high microvessel density and high VEGFmRNA levels (p = 0.006) was found. Furthermore, when studying VEGF expression by immunohistochemistry, we obtained similar results and noted a correlation with VEGFmRNA expression (p < 0.0001). The association between high vascularization, high VEGF levels, and squamous cell histotype suggests the possible role of neoangiogenesis in determining the more aggressive biological behaviour of this type of cancer.     

直肠癌转移与癌组织微血管密度的关系

目的:分析直肠癌组织的微血管密度(MVD)变化及其对癌转移的临床诊断价值。方法:采用免疫组织化学法,对72例手术切除直肠癌组织标本进行微血管标记;在光学显微镜下测定其MVD。将所有患者的临床及追踪随访资料与其手术标本的MVD数据进行统计分析。结果:有转移直肠癌组织的MVD高于无转移者(106.12±32.18vs84.26±27.13),差异有非常显著性意义。结论:直肠癌组织的MVD增加对诊断癌转移有一定的临床价值。     

糖尿病小鼠肾小球微血管密度与VEGF表达的研究

目的: 探讨糖尿病小鼠肾小球微血管密度(MVD)变化及其与血管内皮生长因子(VEGF)表达的关系。方法: 链脲佐菌素诱导小鼠糖尿病6周,定期测量对照组与模型组小鼠体重与血糖变化;常规HE染色观察肾脏形态学变化,采用组织细胞病理分析软件测量肾小球直径、周长及面积;应用免疫组织化学方法检测CD34及VEGF在肾小球的表达,计算MVD值及VEGF表达指数。结果: 与对照组相比,模型组小鼠血糖明显升高(P＜0.01),体重显著降低(P＜0.01),肾小球直径、周长及面积明显增大(P＜0.05);肾小球CD34及VEGF表达明显增强(P＜0.01),肾小球MVD显著增高且与VEGF表达呈显著正相关(r=0.9979,P＜0.05)。结论: VEGF可促进糖尿病小鼠肾小球新生血管生成,VEGF表达上调与糖尿病肾脏病变密切相关。     

上皮性卵巢癌中ALDH1和ABCG2的表达及其与微血管形成的关系

目的:本文旨在探讨上皮性卵巢癌(epithelial ovarian cancer,EOC)中乙醛脱氢酶1(ALDH1)、腺苷三磷酸结合盒转运体G2(ABCG2)蛋白表达及微血管密度(microvessel density,MVD)之间的关系。方法:收集198例EOC术后标本和60例卵巢良性上皮性肿瘤标本,应用免疫组织化学法检测EOC和卵巢良性上皮性肿瘤组织中ALDH1、ABCG2及CD105蛋白(微血管标志物)的表达情况。结果:在EOC和卵巢良性上皮性肿瘤组织中,ALDH1和ABCG2的阳性表达率分别为64.1%、61.6%和8.3%、6.7%;MVD分别为22.6±9.7和5.03±3.35,差异均有统计学显著性(P0.05);ALDH1和ABCG2的表达与EOC的组织学分化、FIGO分期、腹腔器官及淋巴结转移有关(P0.05),MVD与EOC的组织学分化、FIGO分期、腹腔器官及淋巴结转移和腹水形成有关(P0.05);MVD分别与ALDH1和ABCG2的表达呈正相关(P0.01),同时,ALDH1和ABCG2的表达亦呈正相关(P0.01)。Kaplan-Meier生存分析表明,ALDH1和ABCG2的过表达以及MVD的增加均与患者的生存率有关,ALDH1和ABCG2表达阳性及MVD≥23的患者生存率明显低于ALDH1和ABCG2表达阴性及MVD23的患者(P0.05)。多因素分析显示FIGO分期、ALDH1表达、ABCG2表达和MVD是影响EOC根治术后患者预后的独立因素(P0.05)。结论:EOC组织中ALDH1和ABCG2过表达以及MVD与肿瘤的分化程度、转移和预后等因素相关,这些指标的联合检测可能作为对EOC的进展及患者预后判断的重要指标。     

参慈胶囊联合顺铂抑制小鼠Lewis肺癌组织血管生成的机制研究

目的:探讨参慈胶囊及顺铂对Lewis肺癌组织生长及血管生成的抑制作用。方法:将40只C57BL/6小鼠自接种Lewis肺癌瘤株细胞建立实体瘤模型,随机分为参慈胶囊组、参慈胶囊+顺铂组、顺铂组、模型组。模型组予等量生理盐水,其余各组荷瘤小鼠均用药21 d,测量肿瘤重量并计算抑瘤率,采用免疫组化技术检测血管内皮生长因子(VEGF)、微血管密度(MVD)表达情况;采用实时荧光定量PCR(FQ-PCR)技术检测Lewis肺癌荷瘤小鼠癌细胞表达血管内皮生长因子受体-2(KDR)mRNA的情况。结果:(1)在抑瘤率方面,参慈胶囊+顺铂组抑瘤率明显高于其它3组,差异显著(P0.01)。(2)参慈胶囊+顺铂组能降低Lewis肺癌组织VEGF、KDR的表达及MVD的形成,差异显著(P0.01)。结论:(1)参慈胶囊联合顺铂能抑制Lewis肺癌组织生长及血管生成,二者联用具有相加或者协同效应;(2)下调VEGF及其受体KDR的表达,可能是二者联合抗肿瘤血管生成的机制之一。