HER2 protein expression correlates with Lauren classification and P53 in gastric cancer patients

Human epidermal growth factor receptor 2 (HER2) is a key pathological characteristic of gastric cancer (GC). However, the clinical significance of HER2 expression in gastric carcinoma remains controversial. The purpose of this study was to analyze the clinicopathological characteristics of HER2 protein expression, Lauren classification and tumor protein p53 (P53) expression and to evaluate the clinical significance of HER2 protein expression. A total of 176 consecutive patients were prospectively recruited between January 2014 and December 2016 at the Second Affiliated Hospital of Zhejiang University School of Medicine. Histological analysis of the resected tissue was performed for HER2 protein expression using immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Additionally, the expression status of HER2 protein and clinicopathological features were analyzed using the chi-squared (χ²) test. Survival analysis was performed using the Kaplan–Meier method, and differences between the survival curves were determined using the log-rank test. All statistical analyses were conducted using SPSS 22.0 statistical software program (IBM Corp., Armonk, NY). A total of 176 patients with GC were enrolled in this study. Intratumoral heterogeneity of HER2 protein overexpression was observed in 42 of 176 cases with IHC grade 2+, accompanied by FISH positivity and IHC grade 3+. HER2 protein expression was correlated with tumor differentiation (P < .001), Lauren classification (P = .001), Borrmann type (P = .003) and P53 expression (P < .001). HER2 protein positivity was associated with significantly higher overall survival (OS) (P = .038). Overexpression of HER2 protein was observed in 23.9% of the cases and was significantly related to the Lauren intestinal subtype and P53 negative expression. HER2 protein overexpression was independently associated with higher OS.     

Progress of tissue injury in appendicitis involves the serine proteases uPA and PAI-1

Objective. Serine proteases and the matrix metalloproteinases (MMPs) are key factors in the proteolytic cascade and participate in extracellular matrix (ECM) degradation. Fibrinolytic activators and inhibitors may have an effect on inflammatory cells, thereby modulating the inflammatory response. It is reasonable to assume that they may be implicated in the tissue injury in acute appendicitis that subsequently leads to appendix perforation. The purpose of this study was to investigate the expression and distribution of urokinase-type plasminogen activator (uPA) and plasminogen-activator inhibitor type 1 (PAI-1) in appendicitis. Material and methods. Expression of uPA and expression of PAI-1 were measured in tissue specimens from patients with appendicitis (n=30) and in control specimens (n=9), using the quantitative ELISA technique. Distribution of enzymes was studied with immunohistochemistry. The uPA and PAI-1 levels in the subgroups of appendicitis and controls were compared. Results. The overall expressions of uPA and PAI-1 were greater in appendicitis than in control specimens (p <0.001 and p<0.0001, respectively). Expressions of uPA and PAI-1 in phlegmonous (n=15), gangrenous (n=6) and perforated appendicitis (n=9) were all higher than those in controls (n=9), (p<0.01). Moreover, the PAI-1 level was elevated in perforated appendicitis compared with phlegmonous appendicitis (p<0.01). uPA staining was observed in connection with vascular endothelial cells and the serosa stained intensely in specimens from perforated appendicitis. Conclusions. The expression of uPA and especially the over-expression of PAI-1 seem to correlate to the progression of local inflammatory response in acute appendicitis.     

EGFR mutations and HER2/3 protein expression and clinical outcome in Chinese advanced non-small cell lung cancer patients treated with gefitinib

Background  To assess the role of various epidermal growth factor receptor (EGFR) mutations and HER2/3 protein expression as predictive markers of responsiveness to gefitinib therapy in Chinese patients with advanced non-small cell lung cancer (NSCLC). Methods  A total of 106 Chinese NSCLC patients who had failed at least one chemotherapy regimen received gefitinib 250 mg once daily. All the 106 tumors from these patients were screened for mutations in the EGFR exons 18–24, and 84 tumors were studied by immunohistochemistry for HER2/3 expression and correlated with clinical treatment outcome. Results  Patients with EGFR mutations had a significantly higher overall response rate (ORR), longer time to progression (TTP) and overall survival (OS) compared with those with wild-type receptor. No difference in ORR was observed between patients with exon 19 deletion and patients with other EGFR mutations. ORR in HER2-positive patients was significantly higher than in the HER2-negative group, irrespective of EGFR mutational status, and a trend for better ORR was observed for HER3-positive patients. HER2 and HER3 expression levels were not associated with any difference in terms of TTP and OS. Nevertheless, when considering the subgroups of non-responders to gefitinib, median TTP in patients with mutated EGFR was significantly longer than in those with no mutations (8.0 vs. 3.0 months, P = 0.0065). EGFR-mutated patients had no significant difference in ORR, TTP and OS according to HER2 and/or HER3 expression. Conclusions  EGFR mutations are effective predictors for gefitinib efficacy in Chinese patients with advanced NSCLC. HER2 and HER3 expression does not provide any additional information for selecting patients most likely to benefit from gefitinib treatment.     

Transforming growth factor beta-1 in rectal tumour,mucosa and plasma in relation to radiotherapy and clinical outcome in rectal cancer patients

Background Rectal cancer patients are treated with surgery and sometimes radiotherapy. Transforming growth factor-β1 (TGF-β1) acts both as an inhibitor of tumour growth and as a promoter of tumour progression. The aim of this study was to determine the levels of TGF-β1 in tumour tissue, adjacent mucosa and plasma in rectal cancer patients and relate these to the effect of radiotherapy and clinical outcome. Materials and methods One hundred and ten patients scheduled for rectal cancer surgery were included, 49% received pre-operative radiotherapy three-field treatment 5 × 5 Gy. Blood samples and biopsies were taken during surgery and later assayed with enzyme-linked immunosorbent assay for total TGF-β1 and active TGF-β1. Patients were then followed for 3 years. Results Total and active TGF-β1 was higher in tumour tissue compared with rectal mucosa (p < 0.0001). Active TGF-β1 in tumour tissue and rectal mucosa was lower in the irradiated group (p = 0.007; p < 0.0001). Total TGF-β1 was higher in patients with metastases at primary diagnosis (p = 0.005) compared to patients without. In patients who later developed metastases, the levels of active TGF-β1 in plasma were lower (p = 0.004). Local recurrence was associated with lower levels of total TGF-β1 in the rectal mucosa (p = 0.038). Conclusions Higher levels of total TGF-β1 in tumour tissue at surgery may be indicative of distant metastases, and low levels of active TGF-β1 in plasma may indicate a risk of developing secondary metastases. Lower levels of total TGF-β1 in rectal mucosa may influence risk of local recurrence. Measurement of TGF-β1 in rectal cancer patients may be of clinical use in the future.     

uPA、P-选择素蛋白在乳腺癌中的表达及意义

目的 探讨uPA与P-选择素蛋白在乳腺癌浸润、转移中的作用以及二者的关系。方法　采用免疫组化SP法检测uPA与P-选择素蛋白在乳腺纤维腺瘤、乳腺导管内癌及浸润性导管癌组织中的表达。结果 随着乳腺组织的恶性转化、浸润及淋巴结转移,uPA、P-选择素蛋白均呈高表达趋势(P<0.05),二者之间有明显的相关性(r=0.547,P<0.01)。结论 uPA与P-选择素蛋白参与了乳腺癌的浸润及淋巴结转移,联合检测二者,对乳腺癌的预后判断有一定的参考价值。     

Serum HER2 as a predictive biomarker for tissue HER2 status and prognosis in patients with gastric cancer

AIM To investigate the association between serum human epidermal growth factor receptor 2(HER2) extracellular domain(ECD) and tissue HER2 status, and the prognostic value of serum HER2 ECD in patients with gastric cancer.METHODS A total of 239 patients with gastric cancer were enrolled from December 2012 to June 2013. Serum HER2 ECD was determined by chemiluminescent assay, and tissue HER2 status was evaluated by immunohistochemistry and fluorescence in situ hybridization assay. A receiver operating characteristic(ROC) curve was plotted to identify the optimal cut-off value for serum HER2 ECD assay for predicting survival in gastric cancer patients.RESULTS Serum HER2 ECD was significantly correlated with tissue HER2 status(P 0.001), tumor size(P 0.001), and intestinal type of gastric cancer(P =0.021). Serum HER2 ECD levels differed significantly between patients with HER2-positive tissue expression and those with HER2-negative tissue expression. ROC analysis yielded an area under the curve value of 0.79(95%CI: 0.71-0.87, P 0.001), with a sensitivity and specificity of 0.54(95%CI: 0.37-0.70) and 0.93(95%CI: 0.88-0.96), respectively. With a cut-off value of 24.75 ng/m L, high serum HER2 ECD had a negative impact on overall survival of the patients(HR: 1.93, 95%CI: 1.32-4.38, P = 0.006). CONCLUSION Serum HER2 ECD could be a highly specific surrogate biomarker for tissue HER2 status in gastric cancer. Optimal cut-off criteria for predicting survival should be established.     

Intratumoral IGF-I protein expression is selectively upregulated in breast cancer patients with BRCA1/2 mutations

BRCA1/2 mutations predispose to early onset breast and ovarian cancers. The phenotypic expression of mutant alleles, however, is thought to be modified by factors that are also involved in the pathogenesis of sporadic breast cancer. One such protein is IGF-I, one of the strongest mitogens to breast cancer cells in vitro. We have utilized immunohistochemistry to compare the intratumoral IGF-I and IGF-I receptor (IGF-IR) protein expression in 57 BRCA1/2 mutation carriers and 102 matched breast cancer patients without a family history in a nested case-control study. BRCA1 silencing by siRNA was used to investigate the effect of BRCA mutations on IGF-I protein expression. IGF-I protein expression was detected in tumoral epithelium and surrounding stroma, and was significantly upregulated in tumors of BRCA mutation carriers when compared with matched sporadic tumors (epithelial: 87.7% vs 61.8%, P=0.001; stromal: 73.7% vs 34.3%, P<0.001). By contrast, IGF-IR protein expression was confined to malignant epithelium and was unchanged in mutation carriers (52.6% vs 39.2%, P=0.310). While in mutation carriers IGF-IR protein expression was significantly correlated with both epithelial (P=0.003) and stromal IGF-I (P=0.02), this association was less pronounced in sporadic breast cancer (P=0.02 respectively). siRNA-mediated downregulation of BRCA1 in primary human mammary gland cells triggered upregulation of endogenous intracellular IGF-I in vitro. The increased intratumoral IGF-I protein expression in BRCA mutation carriers suggests an involvement of the IGF-I/IGF-IR axis in the biological behavior of breast cancers in this population and could define a potential therapeutic target.     

Therapy of metastatic breast cancer with humanized antibodies against the HER2 receptor protein

The HER2 protein, a member of the epidermal growth factor family, is encoded by the protooncogene c-erbB-2. Its overexpression, occurring in approximately one-third of all breast carcinomas, is associated with a poor prognosis. A humanized mouse antibody against HER2 has been developed by genetic engineering. Here an unspecific human IgG was connected to the recognizing mouse IgG fragment. The allergization typical for allogeneic antibodies does not take place in this context. The effectiveness of this antibody has been confirmed by two international prospective phase III trials that tested it alone and combined with chemotherapy. Both modes of application increased the response rates and the time to progression. Side-effects were rare except for a high rate of cardiac dysfunction when the antibody was combined with anthracyclines. The effectiveness and negligible side-effects of the chimeric antibody against HER2 (Herceptin) render it a valuable tool in the treatment of breast cancer. Received: 8 February 1999 / Accepted: 6 April 1999     

乳腺癌耐药蛋白在非霍奇金淋巴瘤中的表达及意义

目的:探讨非霍奇金淋巴瘤(NHL)患者乳腺癌耐药蛋白(BCRP)的表达及临床意义。方法:采用RT-PCR法检测66例NHL患者新鲜淋巴组织标本BCRP及多药耐药基因(MDR1)的表达,并对所有患者进行随访。结果:66例NHL患者BCRP阳性率45.5%,MDR1阳性率59.1%;不同类型、不同临床分期BCRP阳性率不同;BCRP基因有无表达其缓解率、复发率、5年生存率不同;BCRP+MDR1+、BCRP+MDR1-、BCRP-MDR1+、BCRP-MDR1-4组化疗疗效不同(P<0.05)。结论:本组NHL患者BCRP及MDR1表达率高,是化疗敏感性差的重要原因。     

Pathologic complete response to neoadjuvant anti-HER2 therapy is associated with HER2 immunohistochemistry score in HER2-positive early breast cancer

To evaluate whether pathologic complete response (pCR) to neoadjuvant anti-human epidermal growth factor receptor 2 (HER2) therapy is dependent on the HER2 immunohistochemistry (IHC) score.A total of 181 HER2-positive early breast cancer patients who had received neoadjuvant anti-HER2 therapy were included in this study. Associations were examined between IHC score and tumor pCR status (commonly defined by ypT0+ypN0, ypT0/is+ypN0, or ypT0/is).In trastuzumab-based neoadjuvant-treated patients, ypT0+ypN0 was achieved in 46.0% of patients with HER2 IHC 3+ tumors but only 25.0% of patients with HER2 IHC 2+/fluorescence in situ hybridization (FISH)-positive tumors (P = .016). When pCR was defined as ypT0/is+ypN0 or ypT0/is, 54.7% and 61.3% of patients with HER2 IHC 3+ tumors had a pCR, whereas only 29.5% and 38.6% with HER2 IHC 2+/FISH-positive tumors achieved pCR (P = .004 and P = .008, respectively). The association between dual HER2 blockade and pCR was almost exclusively confined to HER2 IHC 3+ tumors (ypT0+ypN0: 61.9% vs 38.9%, P = .013; ypT0/is+ypN0: 71.4% vs 47.4%, P = .009; and ypT0/is: 81.0% vs 52.6%, P = .002) and was absent in HER2 IHC 2+/FISH-positive tumors. Multivariate logistic regression revealed that HER2 IHC 3+ tumors had a significantly higher probability of achieving ypT0+ypN0 (odds ratio [OR], 0.265; 95% confidence interval [CI], 0.109–0.645; P = .003), ypT0/is+ypN0 (OR, 0.221; 95% CI, 0.094–0.521; P = .001), and ypT0/is (OR, 0.254; 95% CI, 0.111–0.583; P = .001) than HER2 IHC 2+/FISH-positive tumors. A significantly better pCR rate was also found in patients with T1 tumors and patients with dual HER2 blockade.The pCR rate was highly correlated with the HER2 IHC score in neoadjuvant anti-HER2 treatment. The addition of pertuzumab to a neoadjuvant trastuzumab-based regimen improved pCR rates, but there was no significant difference in pCR rates in the IHC 2+/FISH-positive group. This suggests that HER2 IHC scores can predict the effectiveness of treatment.     

Comparative studies of tissue inhibitor of metalloproteinases-1 in plasma, serum and tumour tissue extracts from patients with primary colorectal cancer

OBJECTIVE: We have recently shown that preoperative plasma tissue inhibitor of metalloproteinases-1 (TIMP-1) levels are significantly associated with prognosis of colorectal cancer patients. In addition, we have shown that measurement of plasma TIMP-1 yields information on specificity and sensitivity, which could be used for early detection of colorectal cancer. However, it is not clear whether the increased plasma TIMP-1 levels in colorectal cancer patients are derived from the tumour tissue itself in which it is mainly expressed by the stromal cells located in the vicinity of the cancer cells. The purpose of this study was to examine the association between blood TIMP-1 levels and tumour tissue TIMP-1 levels in colorectal cancer patients. MATERIAL AND METHODS: Preoperative EDTA plasma, citrate plasma and serum, as well as tumour tissue extracts from 49 colorectal cancer patients were measured with a TIMP-1 ELISA that measures total TIMP-1 levels (non-complexed and complexed TIMP-1). RESULTS: The median TIMP-1 level in the 49 tumour extracts was 18.7 ng/mg proteins (range 3.5-152.0 ng/mg protein). The median TIMP-1 value was 133.5 ng/ml (range 58.1-559.0 ng/ml) in EDTA plasma, 130.2 ng/ml (range 57.0-572.0 ng/ml) in citrate plasma and 207.2 ng/ml (range 72.6-828.0 ng/ml) in serum. No significant correlations were found between TIMP-1 content in the tumour extracts and in blood.However, EDTA and citrate plasma TIMP-1 levels (r=0.75; p <0.0001) as well as EDTA plasma and serum TIMP-1 levels (r= .064; p<0.0001) were highly correlated. CONCLUSIONS: The lack of correlation between tumour tissue TIMP-1 and blood levels of TIMP-1 suggests that other sources than the tumour tissue itself may contribute to the increased levels of plasma TIMP-1 in patients with colorectal cancer. However, degradation of cell membranes, rapid secretion into the blood stream and other factors may be responsible for the observed lack of association between TIMP-1 concentrations in blood and tumour tissue extracts.     

The expression of HER2/neu in patients with lung cancer and its associated factors

Background and objectives

Lung cancer is the most common cause of cancer-related death worldwide in both sexes. Evidence suggests the role of genetic factors in lung cancer. Studying of such factors can help understand the cancer prognosis. Overexpression of the human epidermal growth factor receptor-2 (HER2/neu) protein is considered an important prognostic factor in breast cancer, but its role has not been confirmed in lung cancer. Therefore, the present study aimed to investigate the role of its expression in patients with lung cancer.

Materials and methods

In this cross-sectional study, patients aged >18 years who were referred to Afzalipoor Hospital, Kerman, Iran, from 2016 to 2017, and were diagnosed with lung cancer were enrolled into the study if they had a pathological sample of their cancerous lung. Their demographics were recorded, and the sample was sectioned and stained to measure HER2/neu gene expression according to DAKO instructions using heat-induced antigen retrieval (HIER) enzyme marker.

Results

The samples of 100 patients with lung cancer were evaluated (84% men and 16% women) with a mean age of 61.34 years (standard deviation of 12.51 years). HER2/neu expression was significantly associated with the type of cancer and was highest in adenocarcinoma and zero in small cell carcinoma (P < 0.001), but not with patients' sex, age, smoking status and family history of cancer (P > 0.05).

Conclusion

These results emphasized the overexpression of HER2/neu in different types of lung cancer, which can be used further for therapeutic purposes. The results showed that HER2/neu was overexpressed not only in adenocarcinoma but also in other types, like squamous cell carcinoma. Therefore, all subtypes of non-small cell lung carcinoma should be considered for anti-HER2 therapies, and further research is required for small cell lung carcinoma.     

ACE、PAI-1基因多态性与2型糖尿病血浆PAI-1水平相关

20 4例 2型糖尿病患者纤溶酶原激活物抑制物 1(PAI 1)值明显高于 60名正常人 (P <0 .0 5 )。血管紧张素Ⅰ转换酶 (ACE)基因DD型PAI 1水平明显高于其它两型 (均P <0 .0 5 ) ,PAI 1基因 4G/ 4G型PAI 1水平较高 (P <0 .0 5 )。Logistic回归表明 ,ACE基因、PAI 1基因、体重指数、胰岛素敏感指数是血浆PAI 1的危险因素。这些发现提示 ,ACE基因、PAI 1基因多态性可能影响 2型糖尿病患者PAI 1水平。     

Evaluation of angiogenesis, p-53 tissue protein expression and serum VEGF in patients with endometrial cancer

Endometrial carcinoma occurs mostly in post-menopausal women. Classical methods of prognostication, as FIGO stage and histopathologic grade, could be improved by applying additional techniques, utilizing molecular biology and immunochemistry. p-53 tumor suppressor gene, the most commonly mutated gene in human cancers has been shown to play an important role in the biology of gynecologic carcinomas. Angiogenesis, a process of formation of new vessels, being connected to tumors progression and metastatic potential was shown to be linked with tumor suppressor genes expression. The aim of the study was to evaluate relationships between intensity of tumor angiogenesis, serum levels of Vascular Endothelial Growth Factor (VEGF) and tissue p-53 protein expression in endometrial adenocarcinoma. Angiogenic Point's Density (APD) was calculated in hot spots areas using the morphometric appliance. For detection of p53 protein in tumor samples, LSAB + Kit Alkaline Phosphatase (DAKO) was used. VEGF levels were assessed in patient's blood sampled before the operation. Overexpression of p53 protein was found in tumor tissue in 35.2% of cases and mean angiogenic points density was greater in p53 positive cases. Serum levels of VEGF were above the cut off level in 54.5% of patients, in those cases angiogenesis was also elevated. In cases of p53 overexpression, VEGF levels tended to be greater as compared with p53 negative cases. In conclusion, our study demonstrated that angiogenesis was more intensive in p53 positive cases, confirming the hypothesis of tumor suppressor-gene regulation of the process of neovascularization. Serum levels of VEGF were borderline-significantly higher in cases of p53 overexpression, they were also correlated to the angiogenesis. Joint assessment of angiogenesis and tumor suppressor genes expression may contribute to reliable evaluation of the biology of endometrial carcinoma.     

Association of a common genetic variant of the IGF-1 gene with event-free survival in patients with HER2-positive breast cancer

Purpose

Insulin-like growth factor 1 (IGF-1) stimulates mitosis and inhibits apoptosis. High circulating IGF-1 levels are linked with an increased risk of colorectal and breast cancer. Recently, IGF-1 single nucleotide polymorphisms (SNPs), especially variant rs2946834, have been associated with poor clinical outcome in patients with colorectal cancer. In the present study, we aimed to investigate the influence of IGF1 polymorphisms associated with IGF-1 plasma levels on event-free survival in patients with HER2-positive breast cancer.

Methods

The present study included 161 consecutive white patients with HER2-positive breast cancer. Event-free survival was calculated as the time from cancer diagnosis to either relapse or death from any cause. Genomic DNA was extracted from archived formalin-fixed paraffin-embedded tumor tissue samples; five IGF-1 polymorphisms (rs2946834, rs6220, rs1520220, rs5742694, and rs5742678), all associated with IGF-1 levels, were genotyped by SNaPshot assays.

Results

Kaplan–Meier analysis showed a poorer clinical outcome for carriers of the rare allele of SNP rs2946834 (log-rank test, p = 0.020). Concordantly, in univariate Cox regression analyses, the rare allele of SNP rs2946834 was significantly associated with a decreased event-free survival (HR = 3.06 [1.14–8.22]; p = 0.027). Multivariate analysis adjusted for age and tumor stage confirmed this result (HR = 4.02 [1.36–11.90]; p = 0.012). Other investigated polymorphisms of the IGF1 gene were not significantly associated with event-free survival (all p values >0.05).

Conclusions

This study provides first evidence that IGF1 rs2946834 polymorphism is associated with clinical outcome of HER2-positive breast cancer patients. Further studies are warranted to validate these findings.     

乳腺癌组织p16基因甲基化与ER、PR、HER2及p53表达的关系

目的探讨乳腺癌组织中p16基因甲基化与相关受体表达的相关性,进一步提高乳腺癌的诊断水平。方法采用甲基化特异性PCR（MSP）法检测86份乳腺癌组织及40份乳腺癌患者血清中p16基因的甲基化状态;采用免疫组化sP法检测乳腺癌组织中雌激素受体（ER）和孕激素受体（PR）、人类表皮生长因子受体2（HER2）及p53基因表达,分析各指标之间及与乳腺癌之间的关系。结果乳腺癌组织及血清中p16基因甲基化率分别为29．1％、27．5％;15例ER、PR、HER2均为阴性表达者（三阴乳腺癌）,p16基因甲基化率为86．67％（13／16）,非三阴乳腺癌71例,p16基因甲基化率为16．9％（12／71）,P〈0．01。p16基因甲基化与ER、PR蛋白表达呈负相关（r=-0．425、-0．512,P均〈0．05）,与HER2表达呈正相关（r=0．443,P〈0．05）;与p53表达无明显相关性。结论p16基因甲基化是乳腺癌中常见的分子改变,其与ER、PR、HER2联合检测可作为乳腺癌早期诊治及预后判断的重要指标。