Objective: HER-2/neustatus in gastric carcinomas (GCs) has not been studied before in the Egyp- tian population. Materials and Methods: Seventy-three GCs were evaluated for the expression of HER-2/neu and Ki-67 using immunohistochemistry (IHC). Fluorescence in situ Hybridization (FISH) was done for HER2/neu IHC score 2+ cases. Results: Out of the 73 gastric carcinomas, 23 (31.5%) were score 0, 17 (23.3%) were score 1+, 23 (31.5%) were score 2+, and 10 (13.7%) were score 3+. FISH analysis revealed that the HER-2/neu gene was amplified in 11 out of the 23 cases (47.8%) scored 2+ by IHC. Therefore, the overall HER-2/neu positivity rate was 28.8% and it was signifi- cantly associated with higher T-stage and lymphovascular invasion (LVI). The Ki-67 expression rate ranged between 10% and 100%, with 84.9% of the cases (n = 62) featuring high Ki-67 scores. High Ki-67 score was significantly associated with male sex, tumor grade, and number of positive nodes. HER-2/neu protein expression correlated significantly with Ki-67 score. Twenty tumors showed combined HER-2/neu positivity and high Ki-67 score and were significantly associated with higher T-stage and occurrence of LVI, implying a more aggressive behavior. Conclusions: The rate of HER-2/neu positive GCs in our series simulates universal rates, thereby mandating routine evaluation of HER-2/neustatus in all GCs submitted to our laboratory to benefit from trastuzumab therapy. Further studies on a larger number of GCs are required to prove that the concurrent HER-2/neu positivity and high Ki-67 score are markers of worse prognosis.
are no major side effects with Trastuzumab, it is ideal that all patients with gastric carcinoma are tested for HER-2/neu and be given the benefit of it. The laboratories in the hospital should be well equipped with availability both IHC and FISH to test HER-2/neu gene. Patients may develop pimary and acquired resistance to Trastuzumab due to crosstalk between pathways. Research should also be done with combination of targeted therapies to overcome this resistance to improve the outcome of these patients. Beyond all, only a quarter of gastric cancer patients will have amplification of HER-2/neu gene leaving behind the rest 75% of patients without any benefit. Therefore it is time to concentrate on these patients and find out an effective chemotherapeutic or targeting agents which can really change the outcome of gastric cancer treatment. Also more trials are necessary in adjuvant setup to improve the disease free survival in patients who presented at an early stage though equal importance has to be given to prolong the progression free survival of patients who present in advanced disease. Each country should try to identify if any epidemiological factors are influencing the cause and take steps to get rid of those causative agent which will enable us to decrease the incidence of this disease.
Her-2/neu gene amplification is primarily detected by in situ hybridization and uses fluorescence (FISH) to detect the signals. This method is both cumbersome and expen- sive and needs a fluorescence microscope, appropriate fil- ters, and a sophisticated camera; it is therefore not practical as a screening tool. Chromogenic in situ hybridi- zation (CISH) is a recently introduced method, and although it makes use of the in situ hybridization technology of FISH, it also takes advantage of the chromogenic signal detection of IHC that can be detected with the ordinary light microscope and costs one-quarter as much as FISH. CISH is potentially able to detect Her-2/neu gene amplifi- cation and to minimize, if not eliminate, the false positive fraction with the IHC procedure. Here we report an evalua- tion of the CISH assay in St Luke's Medical Center (SLMC), Philippines.
cells under normoxia and hypoxia by Western blot analysis as described in Materials and Methods using an anti-CREB and anti-CREB phosphorylation-specific antibodies. One of three representative Western blots is shown. C. The activity/phosphorylation of the AKT and ERK pathway was determined under normoxic and hypoxic conditions by Western blot analysis using total and phosphorylation-specific antibodies, respectively. The data represent one of three biological replicates. D. Cells were treated under hypoxia with 5 μM LY294002, 100 nM trametinib or 5 μM U0126 for 24 h. The phosphorylation and total expression of CREB, AKT and ERK was analysed by Western Blot. E. Hypoxia-mediated induction of CREB phosphorylation and its dependence on the HER-2/neustatus was determined in HTB122 cells and their HER-2/neu-transformed transfectants (E2A: dominant negative mutation in the HER-2/neu kinase domain; E2: wild-type HER-2/neu) either incubated under normoxia or hypoxia for 24 h, respectively, before Western blot analysis was performed as described in Materials and Methods. The results show one of two independent experiments (up). HTB122, E2 and E2A cells were incubated under normoxic conditions for 24 h before the HER-2/neu cell surface expression was determined using flow cytometry. The data are represented as histograms from one out of two representative experiments. The black area represents the IgG control, while the red defined area is the HER-2/neu-PE staining (down). F. Cells were incubated for 24 h under normoxia or hypoxia and the presentation of HER-2/neu on the cell surface was determined by flow cytometry as described in Materials and Methods using a PE-labelled anti-HER-/neu mAb. The bars represent the MFI of HER-2/neu compared to an IgG control from two independent experiments. G. The effect of inhibition of the HER-2/ neu activity by treatment of parental HER-2/neu + cells with increasing concentrations of lapatinib on CREB phosphorylation was analysed
Currently, treatment of MBC patients with HER-2/neu positive tumors is based on HER-2/neustatus derived from the primary tumor, which was generally removed many years previously and stored as paraffin-embedded blocks. In a 2005 report by Zidan and coworkers  it was pointed out that HER-2/neustatus of the primary tumor may not accurately reflect the HER-2/neustatus of the metastatic tumor, and that this should be taken into account when making treatment decisions. Those investigators demon- strated 14% discordance between primary and metastatic tumors by IHC. Twelve per cent (7/58) of the patients were HER-2/neu positive in the metastatic tumor yet negative in the corresponding primary tumor. Interestingly, three of the seven patients who were HER-2/neu negative in the primary tumor but positive by IHC in the metastatic tumor responded to trastuzumab-based therapy. The references list of that report directed us to several other papers comparing HER-2/ neustatus, as determined using IHC and FISH, between primary breast tumor and metastatic tumor from the same patient. Evidence supporting the observation that a primary breast tumor can be HER-2/neu negative while the metastatic tumor can positive is illustrated below with a few examples. Edgerton and coworkers , employing IHC and FISH, reported 20% discordance between the primary and metastatic tumor, which was due to normal HER-2/neu expression in the primary tumor and HER-2/neu over- expression in the metastatic tumor. Gancberg and colleagues  compared HER-2/neustatus of the primary breast tumor with that of at least one distant metastatic tumor in 107 patients using both IHC and FISH. There was a 6% (6/100) rate of discordance with IHC between the primary and metastatic tumor. In the six cases of discordance, there was greater HER-2/neu staining in the metastatic tumor tissue than in the primary tumor tissue. By FISH analysis, 7% (5/68) of the cases were discordant. Three of the five discordant patient specimens exhibited amplification in the metastatic tumor but not in the primary tumor. It was also reported that if all metastatic tumor sites were included in the analysis, then the HER-2/neu positive metastatic tumors with a corres- ponding negative primary tumor were more frequent than the converse, suggesting that HER-2/neu expression in primary tumors might represent an underestimation of reality. In another study , 80 paired primary tumors and metastatic tumors from the same patients were evaluated for HER-2/neu
Gastroesophageal region cancers (GERCa) had a higher HER- 2/neu positivity rate than other gastric regions . HER-2/neu positivity rate in GERCa was 32.2% according to the ToGa study (the Trastuzumab for Gastric Cancer trial) . We did not determine a significant correlation between HER-2/neustatus and tumor location. Our HER-2/neu positivity rate (R-IHC3+ and R- IHC2+/FISH+) in tumors with cardia location was 8.8% while this rate was 5.3% in cases with non-cardia location. Although gene amplification rates were different between cardia (57.1%) and non-cardia tumors (35%), there was no a statistical significance (p=0.397). HER-2/neu positivity rate in our cases with cardia location was lower than those reported in the literature. Cutsem et al suggested that GERCa were different from GCa with other location in terms of etiological and pathogenetic mechanisms. Higher HER-2/neu positivity rates in GERCa were explained on the basis of their discrete etiopathogenetic characteristics and their predominantly intestinal type morphology [25, 26].
A correlation between serum HER-2/neu levels and tissue HER-2/neustatus in metastatic breast cancer has been examined. A serum ECD concentration of 16 µ g/L showed a sensitivity and specificity of approximately 90% and 80%, respectively, when IHC and FISH were the refer- ence standards. In contrast, another study determined a cut-off of 37 µ g/L with 62% sensitivity and 95% specificity for prediction of the tissue HER-2/neustatus. The research- ers attributed this discrepancy to a difference in assays and proposed using a higher cut-off value for metastatic breast cancer than for diagnosis of primary breast cancer. A very high serum ECD concentration with negative tissue HER-2/ neu could be due to disease recurrence. 27 Trastuzumab is a
elevated risk with a late age regardless of HER-2/neu pro- tein expression . Previous findings suggested an inverse relationship between abortion and HER-2/neu+ breast cancers, while we also found this inverse association but independently of HER-2/neustatus . Interestingly enough, abortion increased risk for HER-2/neu+ tumours only in the premenopausal group of women. Early contra- ceptive use has been positively associated with HER-2/ neu+ breast cancer in two studies [7,8], but our findings were different, revealing a positive association with HER- 2/neu negative tumours. However, because the number of oral contraceptive (and HRT) users in this study was small, this subgroup analysis was hindered by decreased power to detect associations of any magnitude. The slightly protective effect of parity found in the age- adjusted analysis was diminished after logistic regression and did not reveal any association with HER-2/neustatus, in contrast with previous findings . Breastfeeding was associated with increased risks for breast cancer in women with HER-2/neu positive tumours in one study while other investigators reported opposite results [3,9]. Although our study population showed a remarkable lac- tation incidence (almost 80% of the participants) we found no associations at all.
find out the independent factors predicting HER-2/neu amplification. Significant findings: The expression of ER and PR was inversely correlated with HER-2/neustatus in women aged >40 years. By multivariate analysis, as far as the overall groups were concerned, PR, lymph node status and tumor grade were independently associated with HER-2/ neu; Considering the younger age group (≤40), the only predictor for HER-2/neu was the tumor grade; Considering the older age group (>40), tumor grade, PR status, tumor size and lymph node status were associated with HER-2/ neu overexpression. Conclusions: Our data suggest that the association between ER, PR and HER-2/neu is age- related. The negative relationship is only applied for women aged >40 years.
statistically significant (p value <0.001) where the vast majority of ER positive cases (54.76%) were negative for Her-2/neu. Similarly the vast majority of PR positive tumors (320.7%) were Her-2/neu negative; these findings agreed with study in Kirkuk city by  also agreed with two studies . This inverse correlation is one of the reasons why women who have over-expression her-2/neu may be resistant to Tamoxifen . Our results confirmed the suggestion that the presence of ER and PR receptors breast cancer cell lines lead to a strong reduction of her-2/neu protein over-expression. These findings are in agreement with other reports ,  and . Although in other study , they did not find any association between hormonal receptors expression and her-2/neu over-expression which was disagreed with our results it has been suggested that The tumor with positive hormonal receptors has better prognosis and good response to hormonal therapy in contrast to those with no hormonal receptors The relation between hormonal receptor and her-2/neu has been explained by hormone-dependent down regulation of Her-2/neu which involves a very complex molecular interaction. In this interaction Estrogen and its receptor are required to suppress Her-2/neu expression .This explained the Her-2/neu expression in high levels in women with low or absent ER expression. But this had disagreed with study in Iran  which claimed no significant association between hormone receptors and Her-2/neustatus.
Der Nachweis disseminierter Tumorzellen im Knochenmark ist sowohl zum Zeitpunkt der Primärdiagnose als auch während der rezidivfreien Nachsorge mit einer schlechteren Prognose assoziiert und stellt einen unabhängigen Prognosefaktor dar. Diese Tumorzellen sind in 10-40% der Knochenmarkaspirate bei Erstdiagnose nachweisbar. Es wird angenommen, dass diese Tumorzellen dem Primärtumor entstammen und zur Ausbildung von Fernmetastasen fähig sind. Zudem ist eine Änderung des Tumorphänotyps im Verlauf der Erkrankung ein bekanntes Phänomen. Diese einzelnen Tumorzellen können im Knochenmark und im Blut mithilfe von Antikörpern, die gegen Zytokeratine (Marker epithelialer Zellen) gerichtet sind, nachgewiesen werden. Ziel dieser Arbeit war ein Vergleich des HER-2/neu-Status auf dem Primärtumor und auf disseminierten Tumorzellen im Verlauf der Erkrankung.
If HER-2/neu-specific IFN-g producing T cells are involved in HER-2/neu loss and tumor recurrence, we might be able to detect such immune responses in patients with HER-2/neu negative breast cancer, who might have had undetectable HER-2/neu positive pre- malignant tumors in the past, that had lost HER-2/neu expression and progressed to invasive carcinoma under the immune pressure. The fact that 55-75% of patients with premalignant DCIS overexpress HER-2/neu in their tumor lesions and 75% of breast cancers are HER-2/neu negative would suggest the progression of HER-2/neu positive DCIS to HER-2/neu negative breast cancer is only in the tumor clones that express IFN-g Ra. We have already shown that T cell-mediated tumor antigen loss was due to hypermethylation of the neu promoter and loss of neu both at mRNA and protein levels [3,5]. It is likely that DNA methylation may also impact HER- 2/neu gene amplification, as suggested by others . We performed a pilot study accruing 12 breast cancer patients with HER-2/neu negative tumors (HER-2/neustatus: FISH negative) and three breast cancer patients with HER-2/neu positive tumors (FISH positive). We detected HER-2/neu-specific IFN-g producing T cell responses in PBMC of patients with HER-2/neu negative cancers (10 out of 12 patients). Interestingly, there was a direct correlation between the HER-2/neu-specific T cell responses and nuclear localization of IFN-g Ra in the tumors, as an indication of IFN-g responses at the tumor site . A higher IFN-g production in a majority
GADD45 was expressed in 91.6% cases in the study. There was no significant association established between GADD45 expression and response to CTh with doce- taxel and carboplatin (r = 0.263; p = 0.236). But a signifi- cant association was there between GADD45 expression and 2-year disease-free and 2-year overall survival. Those cases which did not express GADD45 had a poor 2-year overall survival of 0% compared to 81% when compared with positive cases (p = 0.005). There is no published report examining the role of GADD45 as a predictive marker of CT response or survival in LA HNSCC till date. In prostate cancer, upregulation of GADD45 levels has been associated with increased sen- sitivity to docetaxel in in vitro studies . They have proposed that GADD45 was epigenetically repressed in prostate cancer and its upregulation may be a potential target for therapeutic strategies . Another experimen- tal study had concluded that tumors in GADD45 defi- cient mice show increased mutation frequency and increased susceptibility to ionizing radiation and to chemical carcinogens . All our cases that expressed GADD45 did so before starting the CTh, and post ther- apy evaluation was not carried out in this study.
The incidence of gastric carcinoma in the year 2011 in RGGGH was 3.26%. Many patients were older than 55 years of age with male preponderance which is similar to several other studies conducted throughout the world. Females showed a younger mean age of incidence compared to males. 29% of cases showed reduced expression of E Cadherin. A significant association was found between reduced expression of E Cadherin and reduced survival. A significant association of EGFR over expression was found with moderately differentiated grade. No significant association was found between EGFR expression and survival. An increased frequency of cases with p53 positivity showed intestinal type of Lauren‘s classification, moderately and poorly differentiated grades and no association between p53 positivity and survival was found. All the cases which showed HER-2/Neu over expression showed T3 level of infiltration, no association with HER- 2/Neu expression and survival was found.
Abstract: Targeting tumors using cancer vaccine therapeutics has several advantages including the induction of long-term immunity, prime boost strategies for additional treatments and reduced side effects compared to conventional chemotherapeutics. However, one problem in targeting tumor antigens directly is that this can lead to antigen loss or immunoediting. We hypothesized that directing the immune response to a normal cell type required for tumor growth and survival could provide a more stable immunotherapeutic target. We thus examined the ability of an antiangiogenesis, Listeria monocytogenes (Lm)-based vector to deliver extracellular and intracellular fragments of the mouse vascular endothelial growth factor receptor-2/Flk-1 molecule, Lm-LLO-Flk-E1, and Lm-LLO-Flk-11 respectively, in an autochthonous model for Her-2/neu + breast cancer. We found that these vaccines could cause
9. Pegram MD, Slamon DJ. Combination therapy with trastuzumab (Herceptin) and cisplatin for chemoresistant metastatic breast cancer: evidence for receptor-enhanced chemosensitivity. Semin Oncol. 1999; 26(4 Suppl 12):89-95. 10. Jimenez RE, Hussain M, Bianco FJ, Jr., Vaishampayan U, Tabazcka P, Sakr WA, Pontes JE, et al. Her-2/neu overexpression in muscle-invasive urothelial carcinoma of the bladder: prognostic significance and comparative analysis in primary and metastatic tumors. Clin Cancer Res. 2001;7(8):2440-2447.
Breast is made up of parenchymal tissues composed of branching ductal system which are radiating from the nipple. 15-20 mammary lobules are usually present in breast parenchyma. Lactiferous duct drains the lobules to the nipple. Lobules are separated from each another by interlobar connective tissue. Before entering the nipple, 15-20 main ducts ,each expands to form a dilated segment known as the lactiferous sinus. Every lobe consists of 30-80 lobules, which are the milk-forming elements of the breast. Each lobules composed of 20-40 terminal units or acini. Lobules are surrounded by intralobar connective tissue. These intralobar connective tissue are hormonally sensitive. The proportion of fibrous, fat and parenchymal tissue vary between individuals and also with weight, menopausal status, genetic factors and the number of live births (9).
detection (RT-PCR, FISH, quantitative PCR), but costs turn them unavailable for broader use in our hospital. Uchino16 detected 2% positive staining of HER2/NEU protein in poorly differentiated gastric carcinomas where as in our study it is found to be 6%, and Tsujimoto17 found no amplification of HER2/NEU gene in undifferentiated type carcinoma by Western Blot. But in our study we found HER2/NEU expression only in poorly differentiated carcinomas .
Morphological analysis. Groups of three WT BALB/c and KO mice were sacrificed at the following times: WT BALB/c mice vaccinated before tumor cell challenge were sacrificed 3 and 7 days after the last shot, and groups of WT BALB/c and KO mice shot when the tumor reach 2 mm in mean diameter were sacrificed each week begin- ning the second week after the first shot until the end of the experiment. For histological evaluation, tissue samples were fixed in 10% neutral buffered formalin, embedded in paraffin, sectioned at 4 µm, and stained with hematoxylin and eosin or Giemsa and the trichrome method. For immunohistochemistry, acetone-fixed cryostat sections were incubated for 30 minutes with antidendritic cells (NLDC 145; Cederlane, Hornby, Ontario, Canada), anti- CD4, anti-CD8a (both from Sera-Lab, Crawley Down, Sussex, United Kingdom), anti-Mac-1 (anti-CD11b/ CD18), anti–Mac-3, anti-Ia (all from Boehringer Mann- heim, Milan, Italy), anti-PMN leukocytes (RB6-8C5, pro- vided by R.L. Coffman), antiasialo GM1 (Wako Chemicals GmbH), antiendothelial cells (mEC-13.324), anti–ELAM- 1 (E selectin; both provided by A. Vecchi, Istituto M. Negri, Milan, Italy), anti–ICAM-1 (CD54), anti–VCAM-1, anti–IL-4, anti–IL-6 (PharMingen, San Diego, California, USA), anti–IL-1β (Genzyme, Cambridge, Massachusetts, USA), anti–TNF- α (Immuno Kontact, Frankfurt, Ger- many), anti–IFN-γ (provided by S. Landolfo, University of Turin), and TGF-β1 (Santa Cruz Biotechnology, Santa Cruz, California, USA) Ab’s. To evaluate the expression of r-p185 neu , paraffin-embedded sections were tested with