Before the decision by the FDA to clear the serum HER-2/
neu test with a standardized cutpoint of 15 ng/ml, many reports used different research assays with various, uncharacterized antibody specificities, calibrators, and cutoff values to measure circulating HER-2/neu level. For instance, in a previous review  it was shown that there were 21 references to the Triton-Ciba-Chiron HER-2/neu assay but 11 different cutoffs, ranging from 5 to 30 units/ml or from 120 or 450 fmol/ml. We found five references to the Nicherei assay, three references to the Calbiochem or the Oncogene Research Products assay, three references to the Dianova assay, and two references to the Bender assay. We could not find any references that described antibody specificities or standardization of these assays, and neither were the findings compared with standardized results. All of the assays mentioned above were available for research use only, which means that the performance characteristics of the assay had not been determined. Unlike FDA-cleared in vitro diagnostic products, there are no manufacturing guidelines for research use only products, which often results in inconsistent findings.
neu protein, such as the tyrosine kinase inhibitor lapatinib, which appears to have clinical activity after failure of tras- tuzumab therapy. 24
Tumors, including metastatic lesions, shed large numbers of tumor cells into the blood circulation, and the presence of circulating tumor cells is of biologic relevance in the metastatic setting. Based on the hypothesis that the pheno- type of circulating tumor cells may reflect the phenotype of metastatic disease, characterization of circulating tumor cells may be useful for reassessment of HER-2/neu status and additional therapeutic cancer biomarkers. However, this option is limited to those patients with detectable circulating tumor cells. Reported positivity rates for circulating tumor cells in metastatic breastcancer patients range from 20%
The HER-2/neu (also known as ERBB2) proto-oncogene, which encodes a trans- membrane growth factor receptor with tyrosine kinase activity, has become an important subject for human cancer re- search during the last decade. The impact of HER-2 amplification and expression on prognosis (1–12) and on the response to cytotoxic (4,13–18) and hormonal (15,19–22) therapies in breastcancer patients have been studied intensively. Studies of HER-2 represent a paradigm of how genetic findings have led to the development of a gene-specific therapy: In September 1998, the U.S. Food and Drug Administration approved trastuzamab (Herceptin), a recombinant monoclonal antibody targeting Her-2, for the treat- ment of metastatic breastcancer. Al- though trastuzumab binds to the Her-2 receptor with high affinity, the mecha- nism of action by which it causes tumor reduction is not understood. Despite the theoretic benefits of such a targeted treat- ment, not all patients respond favorably to trastuzamab treatment in practice. Among the patients with HER-2-positive meta- static breastcancer that is resistant to con- ventional cytotoxic treatment, only about 25% benefit from trastuzamab given in combination with cisplatin (23). The ge- netic features that distinguish the HER-2- positive breast cancers that respond to trastuzamab from those that do not remain unclear. However, it is possible that the extent of HER-2 amplification and/or overexpression in the primary tumor dif- fers from that in the metastases. The HER-2 status of the primary tumor, which is removed from the patient, determines whether or not trastuzamab treatment is prescribed. But trastuzamab works by tar- geting the metastases that remain in the patient. If at least some of the multiple metastases of an HER-2-positive primary breast tumor did not express HER-2, trastuzamab treatment would most likely not affect the course of the disease. A comprehensive, large-scale study compar- ing HER-2 gene copy numbers and pro- tein expression in primary tumors and in multiple different metastases derived from them has not been performed. To gain insight into the patterns of HER-2
Previous studies in Western countries show that TN breastcancer has aggressive clinical and pathologic features, including onset at a young age, advanced stage at diagnosis, high histopathologic and nuclear grade, high mitotic index, higher frequency of unfavorable histopathology, and more distant recurrence. 8–11 In addition, evidence indicates that the prevalence and clinical outcomes of TN breastcancer dif- fer among races. 11 Bauer et al 8 have reported that TN breastcancer is more prevalent among non-Hispanic black women compared with other ethnic groups, who when affected with this subtype had the worst survival. Carey et al 11 also reported that basal-like breast tumors occurred at a higher prevalence among African American women compared with other racial groups. However, there are limited studies of the prevalence, characteristics, and prognosis of TN breastcancer in Asian populations. A recent study of Korean patients indicated that the basal-like subtype, which is positive for one or more of the basal markers and negative for HRs and HER-2/neu, was not associated with a poor prognosis. This study also showed that the survival rate associated with the basal-like subtype does not differ from that of other subtypes, with the exception of the HER2/neu-overexpressing subtype, which has the worst survival rate. 12 In contrast, a recent study of breastcancer patients receiving neoadjuvant chemotherapy showed that TN breastcancer was associated with shorter survival than other subtypes, even though it was associated with a higher response rate. 13
nuclear staining along with cytosolic staining for IFN-g Ra in their tumors.
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 breastcancer, 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 breastcancer 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].
Vikash Kumar et al 73 (2007) studied HER-2/neu oncogene overexpression which was much higher among Indian breastcancer patients 46.3% in comparison to 25-30% in Western literature.
In the present study Estrogen, Progesterone receptor or both were positive in 51.6% cases and both receptors were negative in 48.4% cases (Tab.6). HER-2/neu overexpression was in 42.7% cases (Tab.7). Hence this study is comparable with the studies conducted in the Asian countries. There appears to be a minimal variation in receptor expression; technically this could be explained by differences in technique of evaluation and inter laboratory variations.
sIsH, which directly measures Her-2 gene amplifi- cation, has been added to the armamentarium of tests that can be used to identify patients with Her-2 pos- itive tumor for trastuzumab therapy. The hallmark of Her-2 abnormality in breastcancer is protein over- expression, which in most cases apparently occurs as the result of the corresponding gene amplification 9-11 . Consequently, either IHC assay or FIsH has been ex- plored as a single assay to evaluate respective Her-2 protein or gene status. In many studies, good concor- dance from 89% to 93% was found between the group scored as IHC 3+ and FIsH results 15-18 . nonetheless, some authors consider the IsH to be a more accurate and consistent scoring system for determining Her-2 amplification than Hercep test 19 . In addition, accord- ing to some authors’ experience, IsH studies should be performed on all 3+ and 2+ staining to avoid inap- propriate and toxic treatment 20 . Most data support an algorithm in which IsH testing is restricted to IHC 2+ tumors 21 . IsH is more time consuming and expen- sive than IHC, and is therefore not preferred for pri- mary screening for Her-2 status 22 .
According to the results of our study, the classic histopathological parameters incorporated in postoperative histopathology classification of breast, along with the stage of the disease, and the parameters involved in determining the degree of histological differentiation of primary breastcancer are relevant factors that reflect the values of Nottingham Prognostic Index (NPI). Compared with the HER-2 / neu negative breast cancers, NPI was proved as excellent predictor in classifying HER-2 / neu positive breastcancer that in our series have a high rate of recurrent or metastatic disease. On the other hand, in addition of determining the prognostic groups of HER-2 / neu positive breastcancer, other immunohistochemical parameters must be taken into account that will imply adequate treatment of these patients.
was associated with Comedo carcinoma variant. 3-5 Later in subsequent studies HER-2/neu protein expression associated with breast cancers has been confirmed., further evaluation of this receptor appear warranted to confirm whether this marker can be clinically useful in stratifying patients into low risk groups, which may be followed conservatively, and high risk groups that may require extensive post biopsy surgical procedures to prevent recurrence and to rule out invasive disease with an aggressive phenotype. 6-7 HER-2 /neu gene expression has generally not been specifically implicated in progression or prognosis assessment of lobular carcinoma of breast, but it is strongly associated with increased disease recurrence and a poor prognosis. 8 Over expression is also known to occur in ovarian, stomach, and aggressive forms of uterine cancer. 9 Multiple studies reported a significant co-relation of serum HER-2/neu protein levels with recurrence, metastasis or shortened survival. It also predicts the resistance of breastcancer to chemotherapy and absence of clinical response to hormonal therapy even when estrogen assays are positive. 10 Hence it is concluded that HER-2/neu amplification is an independent predictor of shorter disease with free survival in both node negative node and positive note patients. 11
Further in vivo studies are required to establish the signifi- cance of phosphorylated Akt in relation to HER-2 and sub- cellular localisation of p21 WAF1/CIP1 , with future possibilities that immunodetection of phosphorylated p21 WAF1/CIP1 may imply activation of this pathway. Clinical relevance of HER-2-expressing tumours with cytoplasmic p21 WAF1/CIP1 is emphasised by the independent prognos- tic significance of both parameters. Prognostic signifi- cance of cytoplasmic p21 WAF1/CIP1 at 5 years retains its predictive potential for OS and RFS at 9 years. The com- bined evaluation of cytoplasmic p21 WAF1/CIP1 and HER-2 positively did not increase prognostic significance in this study, due to limited patient numbers in the respective subsets. As previously reported, nuclear p21 WAF1/CIP1 expression was generally low and provided no prognostic information [17,33,35]. Future studies will be required to address what are the best markers.
Gastric carcinoma is one among the top five causes of cancer in both males and females. It is also listed among the top cancer related mortalities. Despite the advancements in the treatment of gastric cancer, none has shown to improve the survival significantly. There was no additional improvement in overall survival with addition of Epirubicin or Docetaxol to Cisplatin and 5FU combinations although the response rate was better. The role of targeted therapy has recently come into play after the establishment of the role of HER-2/neu over-expression in gastric malignancies. Trastuzumab ,an anti HER-2/neu antibody ,in combination with Cisplatin and 5FU in advanced gastric carcinoma patients has shown to improve overall survival in TOGA trial This study was undertaken to find out the prevalence and the clinicopathological correlation in HER-2/neu over –expression in gastric carcinoma patients.
1 Cancer Immunology and Immunotherapy Center, Saint Savas Cancer Hospital, 171 Alexandras Avenue, 115 22 Athens, Greece; 2BreastCancer Clinic, Saint Savas Cancer Hospital, 171 Alexandras Avenue, 115 22 Athens, Greece; 3 Abteilung Fuer Physicalische Biochemie des Physiologisch-Chemisches Institut der Universitat, 4 Hoppe-Seyler Strasse, 72076 Tuebingen, Germany
HER-2/neu oncoprotein contains several major histocompatibility complex class I-restricted epitopes, which are recognised by cytotoxic T lymphocyte (CTL) on autologous tumours and therefore can be used in immune-based cancer therapies. Of these, the most extensively studied is HER-2(9 369 ). In the present report, we used dendritic cells pulsed with HER-2(9 369 ) to stimulate, in the presence of IL-7 and IL-12, the production of IFN-g by patients’ CTL detected by the enzyme-linked immunosorbent spot-assay.
These data additionally support the use of combination therapies directed against both HER-2/neu and VEGF for treatment of breast cancers that exhibit HER-2/neu overexpression. However, this study included small number of cases, so further studies with larger sample size are required in this area to conclude an unequivocal association between HER-2/neu and VEGF in breastcancer and its implication in guiding therapy against HER-2/neu overexpressing tumors.
In conclusion, the positive expression of these biomar- kers is associated with biologically aggressive tumors and poor prognostic profile. Although the samples were taken from an area where the exposure to depleted ura- nium is a risk, the incidence of co-expression of both p53 and HER-2/neu markers does not differ from simi- lar cancer samples in areas that have not been exposed to depleted uranium, though, the greater immunoex- pression of Her-2/neu in breastcancer in this popula- tion with risk for DU exposure, compared with findings on other populations not at risk, requires further inves- tigation as it may reflect the possible role of DU in the induction or acceleration of network signaling between different Her-2 receptors. New lines of treatment which includes genetic modulation of the signaling pathway of both genes should be considered in patients’ medical follow up. Unfortunately for DU, knowledge of the exposure time, dose absorbed, route, length of exposure and its health consequences on the Iraqi population is still lacking. This is chiefly due to restricted access of scientists required to conduct such study and should form the basis for future investigations.
An inverse association between ER and PR with HER-2/
neu has been observed by others [10, 13, 14].
In our study, ER negativity was more strongly associated with HER-2/neu negativity. It was the same with PR negativity. However our study has some limitations. Malaysian population represent a multiracial community with Chinese, Malays and Indians as major racial groups and this biological diversity may be responsible for differences in ER, PR and HER-2/neu expression. Furthermore our results may have been affected by the use of immuno histochemistry analysis of HER-2/neu. Therefore more studies are needed from Malaysia to verify our findings.
conferred by adjuvant chemotherapy or endocrine therapy was examined in randomized studies [2,3,8,9,35–38].
Because adjuvant therapies may positively impact on outcome in only around 15%, the costs of these therapies make their routine application in all NNBC patients inap- propriate . For this reason, predictive factors that accu- rately define subgroups of NNBC patients who may benefit from adjuvant systemic therapy would be a great advantage.
1 Department of Pathology, Karmanos Cancer Institute, Wayne State University School of Medicine, Detroit, Michigan, United States of America, 2 Division of Hematology/
Oncology, Karmanos Cancer Institute, Wayne State University School of Medicine, Detroit, Michigan, United States of America
Herceptin failure is a major clinical problem in breastcancer. A subset of breastcancer patients with high HER-2/neu levels eventually experience metastatic disease progression when treated with Herceptin as a single agent. Mechanistic details of development of this aggressive disease are not clear. Therefore, there is a dire need to better understand the mechanisms by which drug resistance develops and to design new combined treatments that benefit patients with aggressive breastcancer and have minimal toxicity. We hypothesized that 3, 39-diindolylmethane (DIM), a non-toxic agent can be combined with Herceptin to treat breast cancers with high levels of HER-2/neu. Here, we evaluated the effects of Herceptin alone and in combination with DIM on cell viability, apoptosis and clonogenic assays in SKBR3 (HER-2/neu-expressing) and MDA-MB- 468 (HER-2/neu negative) breastcancer cells. We found that DIM could enhance the effectiveness of Herceptin by significantly reducing cell viability, which was associated with apoptosis-induction and significant inhibition of colony formation, compared with single agent treatment. These results were consistent with the down-regulation of Akt and NF-kB p65. Mechanistic investigations revealed a significant upregulation of miR-200 and reduction of FoxM1 expression in DIM and Herceptin-treated breastcancer cells. We, therefore, transfected cells with pre-miR-200 or silenced FoxM1 in these cells for understanding the molecular mechanism involved. These results provide experimental evidence, for the first time, that DIM plus Herceptin therapy could be translated to the clinic as a therapeutic modality to improve treatment outcome of patients with breastcancer, particularly for the patients whose tumors express high levels of HER-2/neu.
Rak dojke je najpogostejša maligna bolezen pri ženskah. Tako kot povsod po svetu tudi v Sloveniji v zadnjih letih opažamo stalen porast incidence, ki že presega 1000 novih bolnic na leto (1). Na potek bolezni in učinkovitost zdravljenja karcinoma dojk vplivajo številni dejavniki, predvsem biološki. Posebno vlogo pridobiva izraženost proteina her-2 (humani epidermalni receptor 2 za rastni faktor) in pomnožitev gena HER-2. Protein her-2 je čezmerno izražen oziroma gen HER2 je pomnožen pri okoli 20 % bolnic z rakom dojke. Na Onkološkem inštitutu v Ljubljani določamo HER-2 status v tkivu raka dojk od leta 2000, vendar na lastni populaciji do sedaj še ni bilo opravljene analize o vplivu tega dejavnika na potek bolezni. Pričujoča raziskava je prva analiza preživetja bolnic z HER2 pozitivnim rakom dojk v našem prostoru. Her-2, imenovan tudi c-erbB-2 ali neu je transmembranski protein s tirozin kinazno aktivnostjo. V majhnih koncen- tracijah je izražen v duktalnem epiteliju dojke in v mnogih drugih normalnih epitelijih. V večji meri je pomnožen pri petini do četrtini invazivnih rakov dojke. V mnogih raziskavah so dokazali, da je prekomerna izraženost her-2 neodvisen napovedni dejavnik preživetja, posebno pri bolnicah z rakom dojk in zasevki v pazdušnih bezgavkah. Prekomerno izraženi protein napoveduje dober odziv na antraciklinsko kemoterapijo, hkrati pa slab odziv na tamoksifen celo pri bolnicah s pozitivnimi hormonskimi receptorji (2). V zadnjem času je bilo razvito tarčno zdravilo, ki zavre receptor HER-2, trastuzumab. Določanje prekomerno izraženega proteina her-2 ali pomnožitve gena HER2 je postala metoda za izbiro tistih bolnic z metastatsko in zgodnjo boleznijo, ki imajo dobrobit od zdravljenja s trastuzumabom, humaniziranim protitelesom proti proteinu HER-2. Zato danes določamo HER2 status z izraženostjo proteina in/ali pomnožitvijo gena HER-2 v vsakem na novo odkritem karcinomu dojke. Status HER-2 lahko določamo na dva načina. Število kopij gena HER-2 lahko analiziramo s fluorescentno hibridizacijo in situ (FISH), izraženost membranskega proteina her-2 pa z imunohistokemijo. Obe metodi lahko v praksi izvajamo s standardnimi parafinskimi bloki.
tumours. Most of the efforts have therefore been focused on the cellular immune response and the identification of antigens recognized by human T lymphocytes.
Few TAAs have been identified for breastcancer, and they generally correspond to differentiation antigens or over- expressed normal proteins. Potential new target antigens have recently been described for breastcancer , but most of them are expressed on only a small percentage of breast cancers. One of the first TAAs described for breastcancer was HER2/neu, a 185 kDa transmembrane glyco- protein and member of the epidermal growth factor recep- tor family. Amplification and/or overexpression of HER2/neu have been reported in 10–40% of primary breast cancers, and also in ovarian, renal, gastric and colorectal carcinomas. In this review, we shall focus on the identification and application of HER2/neu peptides as tumour vaccines for T cell recognition.
Triplex-forming oligonucleotides (TFOs) have been shown to bind to target DNA sequences in several human gene promoters such as the c-myc oncogene, the epidermal growth factor receptor, and the dihydrofolate reductase genes. TFOs have been shown to inhibit
transcription in vitro and gene expression in cell culture of the c-myc and other genes. The HER-2/neu oncogene, which is overexpressed in breastcancer and other human