Bruno Henrique Bressan da Costa, Aline Paixão Becker, Luciano Neder, Paola Gyuliane Gonçalves, Cristiane de Oliveira, Allan Dias Polverini, Carlos Afonso Clara, Gustavo Ramos Teixeira, Rui Manuel Reis, Lucas Tadeu Bidinotto
J Pathol Transl Med. 2022;56(4):205-211. Published online June 15, 2022
Background Despite the advances in glioblastoma (GBM) treatment, the average life span of patients is 14 months. Therefore, it is urgent to identity biomarkers of prognosis, treatment response, or development of novel treatment strategies. We previously described the association of high epidermal growth factor-like domain multiple 7 (EGFL7) expression and unfavorable outcome of pilocytic astrocytoma patients. The present study aims to analyze the prognostic potential of EGFL7 in GBM isocitrate dehydrogenase (IDH)-wildtype, using immunohistochemistry and in silico approaches.
Methods Spearman’s correlation analysis of The Cancer Genome Atlas RNA sequencing data was performed. The genes strongly correlated to EGFL7 expression were submitted to enrichment gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Additionally, EGFL7 expression was associated with patient overall survival. The expression of EGFL7 was analyzed through immunohistochemistry in 74 GBM IDH-wildtype patients’ samples, and was associated with clinicopathological data and overall survival.
Results In silico analysis found 78 genes strongly correlated to EGFL7 expression. These genes were enriched in 40 biological processes and eight KEGG pathways, including angiogenesis/vasculogenesis, cell adhesion, and phosphoinositide 3-kinase–Akt, Notch, and Rap1 signaling pathways. The immunostaining showed high EGFL7 expression in 39 cases (52.7%). High immunolabelling was significantly associated with low Karnofsky Performance Status and poor overall survival. Cox analysis showed that GBMs IDH-wildtype with high EGFL7 expression presented a higher risk of death compared to low expression (hazard ratio, 1.645; 95% confidence interval, 1.021 to 2.650; p = .041).
Conclusions This study gives insights regarding the genes that are correlated with EGFL7, as well as biological processes and signaling pathways, which should be further investigated in order to elucidate their role in glioblastoma biology.
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Background In this study, we investigate the expression of markers of angiogenesis and microvessel density (MVD) in cases of microcystic, elongated and fragmented (MELF) pattern, with its prognostic role in the survival of endometrioid endometrial adenocarcinomas (EA) patients.
Methods In this study, 100 cases of EA, 49 cases with MELF pattern and 51 without, were immunohistochemically stained for galectin-1, vascular endothelial growth factor (VEGF), and MVD. Morphometry and statistical (univariate and multivariate) analyses were performed to assess overall survival (OS) and disease-free survival.
Results The expression of VEGF (p<.001) and galectin-1 (p<.001), as well as MVD area (p<.001) and number of vessels/mm2 (p<.050), were significantly higher in the +MELF pattern group compared to the –MELF group. A low negative correlation between MELFpattern and the number of days of survival (p<.001, r=–0.47) was also found. A low positive correlation of MELF-pattern with galectin-1 expression (p<.001, r=0.39), area of vessels/mm2 (p<.001, r=0.36), outcome of EA (p<.001, r=0.42) and VEGF expression (p<.001, r=0.39) suggests potential pathological relevance of these factors in the prognosis of EA. A univariate survival analysis indicated a role for all parameters of survival. Multivariate Cox proportional hazard regression analysis revealed that only area of vessels/mm2 (hazard ratio [HR], 1.018; 95% confidence interval [CI], 1.002 to 1.033), galectin-1 (HR, 1.049; 95% CI, 1.025 to 1.074) and VEGF (HR, 1.049; 95% CI, 1.022 to 1.077) play key roles in OS.
Conclusions This study reports an increase in MVD, VEGF and galectin-1 expression in EA with MELF pattern and suggests that MELF pattern, along with the angiogenic profile, may be a prognostic factor in EA.
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Background MicroRNAs (miRNAs) are short, non-coding RNAs that mediate post-transcriptional gene regulation. They are commonly deregulated in human malignancies, including non-small cell lung cancer (NSCLC). The aim of this study is to investigate miRNA expression in T790M-mutated NSCLC resistant to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors.
Methods Six cases of resected NSCLC harboring the T790M mutation were examined. We performed miRNA time polymerase chain reaction (PCR) array profiling using EGFR T790M-mutated NSCLC and L858R-mutated NSCLC. Once identified, miRNAs that were differentially expressed between the two groups were validated by quantitative real-time polymerase chain reaction (qRT-PCR).
Results miRNA PCR array profiling revealed three up-regulated miRNAs whose expression levels were altered 4.0-fold or more in the EGFR T790M mutation group than in the L858R group: miR-1 (fold change, 4.384), miR-196a (fold change, 4.138), and miR-124 (fold change, 4.132). The three differentially expressed miRNAs were validated by qRT-PCR, and they were found to be overexpressed in the T790M group relative to L858R group. In particular, expression levels of miR-1 and miR-124 were significantly higher in the T790M group (p-value of miR-1 = .004, miR-124 = .007, miR-196a = .096).
Conclusions MiR-1, miR-124, and miR-196a are overexpressed in EGFR T790M mutated NSCLC.
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BACKGROUND Epidermal growth factor receptor (EGFR) is frequently expressed in the invasive front of colorectal cancer (CRC), but its clinicopathologic significance remains unclear. We investigated the clinical value of the EGFR expression at the invasive front of CRC. METHODS We performed an immunohistochemical analysis in order to examine the expression and distribution of EGFR in 214 cases of CRC. The EGFR status was considered positive when > or =1% of the tumor cells had membranous staining. RESULTS Overall, an EGFR expression was observed in 144 (67%) cases and it had no significant relationship with the clinicopathologic parameters. However, an EGFR expression at the invasive front was correlated with lymphatic invasion, lymph node metastasis and a high level of serum carcinoembryonic antigen (p = 0.028, p = 0.043, and p = 0.045, respectively). For the budding-positive CRCs liver metastases were found in the cases with an EGFR expression at the budding, but no liver metastasis occurred in the EGFR negative cases at the budding (p = 0.030). CONCLUSIONS An EGFR expression at the invasive front has clinicopathologic significances in patients with CRC. An EGFR expression at tumor cell budding is a pathologic marker that suggests the high potential for liver metastasis in CRC.
Transitional cell carcinoma(TCC) of the urinary bladder shows marked heterogeneity in biological behaviors. Evidence has accumulated that biological markers may provide significant information to predict the potential aggressiveness of TCC. We have assessed the expression of the epidermal growth factor receptor (EGF-R), c-erbB-2 and p53 proteins in 56 cases of TCC to investigate the prognostic significance of differential expression of these oncoproteins using an immunohistochemical method. We analysed the expression patterns of these oncoproteins according to tumor stage and grade. And we assessed the probability of progression-free survival in stage T1 tumors according to their expressions. Positive rates of EGF-R (>+3 staining intensity), c-erbB-2 (intense membrane staining) and p53 proteins (>20% positive cells) were 73.2%, 37.5% and 42.9%, respectively. Invasive tumors had significantly higher positive rates of all three factors than did superficial tumors (p<0.005 for EGF-R and c-erbB-2, p<0.05 for p53). High grade tumors had significantly higher positive rates of c-erbB-2 and p53 proteins (p<0.005). In superficial tumors, T1 tumors had higher positive rate of p53 protein compared with Ta tumors (p<0.05). Twelve cases of superficial tumors (34.3%) were positive for EGF-R and negative for c-erbB-2 and p53 proteins. Nine cases of superficial tumors(25.7%) were negative for all three factors. In invasive tumors, however, 42.5% of the cases were positive for all three factors. The overexpression of p53 protein was the only useful marker to predict the rapid progression in stage T1 tumors (p<0.05, log-rank test).
These results suggest that the differential overexpression of EGF-R, c-erbB-2 and p53 proteins could be useful to depict tumor aggressiveness of TCC of the urinary bladder.
And, the overexpression of a p53 protein may be a useful marker to predict the possibility of rapid progression in stage T1 tumors.
Many studies have shown that angiogenesis plays an important role in the growth, the progression, and the metastasis of a solid tumor. The vascular endothelial growth factor(VEGF) is thought to be a selective mitogen for endothelial cells.
Twenty eight advanced gastric carcinomas and twenty early gastric carcinomas were investigated by staining with polyclonal antibody against the VEGF. Correlation between the expression of the VEGF and the clinicopathologic features of gastric carcinoma were studied. The VEGF was mainly localized to the cytoplasm of carcinoma cells. Normal gastric foveolar epithelium was not immunoreactive, but some endothelial cells were weakly immunoreactive with an anti-VEGF antibody. Expression of the VEGF was significantly higher in advanced gastric carcinoma than in early gastric carcinoma (p=0.003). Expression of the VEGF was correlated with the depth of tumor, the lymph node metastasis, and the stage (p<0.05). The VEGF positivity was significantly higher in moderately and poorly differentiated gastric carcinoma than in well differentiated gastric carcinoma. The VEGF scores of the metastatic foci in the lymph nodes were higher than that of the primary tumors, which were followed by deep and superficial portions of the primary tumors in a descending order (p<0.05). In summary, the expression of the VEGF may be associated with progression and metastasis of a gastric carcinoma and may also be a good prognostic factor in a gastric carcinoma.
Growth stimulatory/inhibitory factors and their receptors are the important mediators of control of epithelial cell proliferation and differentiation. The aim of this study was to observe the distribution of epidermal growth factor receptor (EGFR) and transforming growth factor-beta1 type II receptor (TbetaRII) during carcinogenesis of oral squamous cell carcinoma (OSCC). Formalin fixed, paraffin embedded tissue from 25 oral leukoplakias (OL) and 15 OSCC was immunostained by avidin-biotin complex method. In OSCC, the carcinomatous area and the adjacent dysplastic/ hyperplastic area were examined. In OL, the hyperplasia and the epithelial dysplasia were examined. Monoclonal anti-EGFR Ab and polyclonal anti-TbetaRII Ab were applied. EGFR was mainly expressed in the basal layer and was increased with epithelial dysplasia in OL. TbetaRII was not detected in the basal cell layer and dysplastic area in OL. In contrast, the dysplastic area adjacent to OSCC showed positivity in the entire layer including the dysplastic area. In all cases of OSCC, both EGFR and TbetaRII showed positive reactions. EGFR was increased with the progression to the malignancy, and the expression pattern of TbetaR II was altered to be positive in the basal cell layer with progression to malignancy. These results suggest that the expression of EGFR appeared to be an early event and TbetaR II may be related to malignant transformation during oral carcinogenesis. The expression pattern of EGFR and TbetaR II may contribute to predict the risk of the development of carcinoma in oral premalignant lesions.
Angiogenesis is an early event in tumorigenesis and facilitates tumor progression and metastasis. This study was performed to evaluate the expression of vascular endothelial growth factor (VEGF), phospholipase C-gamma1 (PLC-gamma1) and Ki-67, and to assess the relationship between them in cervical squamous cell neoplasia. The materials were fifty cervical squamous cell lesions, consisted of thirty HSIL (6 moderate dysplasia, 11 severe dysplasia, 13 carcinoma in situ), and twenty invasive squamous cell carcinoma (ISCC) cases. Immunohistochemical stain for VEGF, PLC-gamma1 and Ki-67 were done. Expression rate of VEGF was significantly higher in ISCC than in HSIL (p=0.012). PLC-gamma1 expression was significantly higher in ISCC than in HSIL (p=0.004).
Ki-67 labelling index was significantly higher in ISCC than in HSIL (p=0.001) and higher in VEGF-positive tumors than in VEGF-negative tumors (p=0.018), but there were no significant differences between PLC-gamma1 expression and Ki-67 labelling index (p>0.05), and between PLC-gamma1 and VEGF (p>0.05). This study suggests that PLC-gamma1 and VEGF may play an important role in tumor cell proliferation and invasion, and these may be a useful marker to predict the possibility of invasion in cervical cancer.
Angiogenesis is a critical factor in the progression of solid tumors, including cervical cancers. The mechanisms responsible for angiogenesis in uterine cervical neoplasia are not well defined. To determine the relationship between angiogenesis and the expression of vascular endothelial growth factor (VEGF) in the cervical neoplasia, the author studied 63 cases of the cervical neoplasia diagnosed between the years 1993 to 1997 at Pusan National University Hospital. The expression of VEGF was semiquantitatively analyzed in paraffin sections by immunohistochemical method.
Histologic sections immunostained for factor VIII-related antigen were evaluated for microvessel density. Increased expression of VEGF and microvessel counts was significantly correlated with depth of invasion. Increased microvessel counts were also significantly associated with increased VEGF expression. These results suggest that VEGF is an important angiogenic factor and associated with progression of the cervical neoplasia.
Recent investigations have revealed that autocrine growth factors and their receptors are closely related and play an important role in controlling cancer cell growth. We performed an immunohistochemical study on the expression of epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), epidermal growth factor receptor (EGF-R), c-erbB-2, and PCNA labelling index in 60 cases of human gastric carcinomas. TGF-alpha was detected in 38 cases (63.3%), EGF in 26 cases (43.3%), EGF-R in 44 cases (73.3%), and c-erbB-2 in 18 cases (30%). These growth factors, EGF-R and c-erbB-2, were found more often in advanced gastric cancers. The PCNA labeling index was significantly higher in tumors with the expression of EGF-R or c-erbB-2. Tumors with simultaneous expression of EGF, TGF-alpha, EGF-R and c-erbB-2 was associated with a high PCNA labeling index. A correlation was observed between the synchronous expression of growth factors and its receptors and histological differentiation. The results suggest that the expression of EGF, TGF-alpha, EGF-R and c-erbB-2 are closely related and plays an important role in the growth and progression of human gastric carcinoma.
Angiogenesis is essential for tumor growth and metastasis, however, the prognostic value of neovascularization is undetermined. The aim of this study is to evaluate the prognostic significance of microvessel density (MVD) and vascular endothelial growth factor (VEGF) expression in breast carcinomas. An immunohistochemical stains for CD 31 (DAKO) to estimate MVD and VEGF (Santa Cruz) were done on 40 cases of invasive breast carcinoma.
MVD was calculated as an average count of vessels per 200 power field in the most vascularized areas. VEGF expression was interpreted according to staining intensity and number of positive cells.
Mean MVD was 35, and MVD was not correlated with lymph node metastasis or histologic grade, but high MVD (mean MVD>35) showed an increasing tendency in cases with larger size, negative ER/PR, and positive cathepsin D. All of the cases showed VEGF expression, but VEGF expression was not correlated with tumor size, histologic grade, lymph node metastasis, ER/PR status, and cathepsin D expression.
These results suggest that MVD and VEGF expressions are not reliable prognostic factors.
Angiogenesis is an essential process in tumor growth and metastasis. VEGF has been considered a leading candidate inducing tumor angiogenesis. VEGF expression was significantly correlated with clinical stage, lymph node matastasis, and prognosis of cancers of various parts of body. However, little has been known about the correlation between VEGF expression and clinicopathologic parameters in oral squamous cell carcinoma. The aim of this study was to correlate VEGF expression with the clinicopathological parameters and microvessel density.
Forty six oral squamous cell carcinomas were analyzed using immunohistochemical method with primary antibodies to VEGF and CD31. VEGF expression was detected in 33 (71.7%) of the 46 cases. The microvessel density was significantly correlated with VEGF expression (P=0.002). There was no correlation between microvessel density and tumour size, clinical stage, and lymph node metastasis, respectively. VEGF expression did not correlate with the histological grade of tumour differentiation, tumour size, and clinical stages. The VEGF-positive rate seemed to be higher in patients with cervical lymph nodal metastasis than in those without it, but it was not statistically significant. In conclusion, the overexpression of VEGF in the oral squamous cell carcinoma seemed to be associated with a more aggressive course of the disease.
Further study is necessary to define the role of VEGF in oral squamous cell carcinoma.
Angiogenesis is an essential requirement for development, progression, and metastasis of malignant tumors. Vascular endothelial growth factor (VEGF) is one of the important angiogenic factors. Recently the role of angiogenesis has been known in premalignant lesions. This study was performed to determine whether the angiogenesis and VEGF expression were increased in association with histological grade of cervical intraepithelial neoplasia (CIN) and to see the relationship between the angiogenesis and VEGF.
Immunostainings for factor VIII and VEGF were performed on 52 cases of cervical neoplasia (12 cases of CIN I, 11 cases of CIN II, 15 cases of CIN III, 7 cases of microinvasive squamous cell carcinoma, and 7 cases of invasive carcinoma) and 5 cases of normal cervix. The results showed a significant increase of microvessel count from normal cervix through CIN grades to invasive squamous cell cacinoma. VEGF expression was increased in proportion to the CIN grades. There was no significant correlation between microvessel count and VEGF expression. In conclusion, the tumor angiogenesis is an early event in tumorigenesis of uterine cervix. In addition, no significant relationship between the microvessel count and VEGF expression in CIN suggests the possibility of other growth factors affecting mainly angiogenesis of premalignant lesion of uterine cervix.
Jinyoung Yoo, Kyungji Lee, Ji Han Jung, Byoung Yong Shim, Sung Hwan Kim, Deog Gon Cho, Myeong Im Ahn, Chi Hong Kim, Kyu Do Cho, Hoon Kyo Kim, Seok Jin Kang
BACKGROUND Gefitinib is an EGFR tyrosine kinase inhibitor that has shown dramatic effectiveness in a subset of non-small cell lung cancer (NSCLC) patients. We evaluated the response rate to gefitinib, and the significance of the EGFR and HER2/neu status as predictive markers of the tumor response. METHODS The EGFR and HER2/neu protein expressions, as determined by immunohistochemistry (IHC) and gene amplification via chromogenic in situ hybridization (CISH), were analyzed in biopsy specimens from 46 patients with advanced NSCLC. After their failure with the first-line treatment, all the patients had received gefitinib treatment. RESULTS A partial response (PR) was achieved in 8 patients (17.4%). An EGFR overexpression was detected in 80.4% (37/46) of the tumors, and this was observed exclusively in patients with a PR (100% vs 75.3%, respectively; p=0.076).
EGFR gene amplification was present in 47.8% of the tumors (22/46). HER2/neu was overexpressed in 13%(6/46) and it was amplified in 17% (7/46). The overall survival was prolonged in the female patients (p=0.007), and in patients with T1 and T2 disease (p=0.039), adenocarcinoma (p=0.010), a PR (p=0.022), an EGFR IHC+ status (p=0.033), an EGFR IHC+/CISH+ status (p=0.010), or an EGFR+/HER2/neu+ status (p=0.030). On multivariate analysis, gender, T disease and EGFR IHC/CISH remained the significant predictors of survival. CONCLUSIONS Gefitinib showed a modest effect for the patients with chemotherapy-refractory advanced NSCLC. A combination of EGFR IHC and CISH might be important for identifying those patients who are most likely to benefit from gefitinib therapy.
BACKGROUND Vascular endothelial growth factor-C (VEGF-C) is a novel growth factor that regulates lymphangiogenesis and/or angiogenesis via binding to the vascular endothelial growth factor receptor-3 (VEGFR-3) or VEGFR-2. Recent studies have suggested that VEGF-C may play a role in lymph node metastasis. This study was conducted to examine whether the expression of VEGF-C is associated with the clinicopathologic parameters, and especially lymph node metastasis, of invasive ductal carcinoma. METHODS Immunohistochemical staining was performed for VEGF-C and CD31 in the surgically resected specimens from 83 patients with invasive breast carcinoma. RESULTS Of the 83 breast carcinomas, 61 (74%) cases showed cytoplasmic VEGF-C imunoreactivity. VEGF-C expression was associated with lymph node metastasis (p=0.03), but it did not correlate with tumor size, the histologic grade, and the presence of estrogen receptor or progesteron receptor. The mean microvessel density in the cases without VEGF-C expression was 51.9+/-30.1 and it was 72.9+/-33.0 in the cases with 2+ expression for VEGF-C (p=0.07). CONCLUSIONS This study suggests that VEGF-C expression may have an association with lymph node metastasis in the patients with breast carcinoma.