Medullary Cancer Research Articles

Angiopoietin 1 (Ang-1), angiopoietin 2 (Ang-2) and Tie-2 (a receptor tyrosine kinase) concentrations in peripheral blood of patients with thyroid cancers

There are limited and conflicting studies investigating the role of the angiopoietin family in human thyroid cancer development and progression. We have investigated cytokines angiopoietin-1, -2 and their receptor (Tie-2), known to be involved in angiogenesis, in the serum of 52 thyroid cancer patients (21 cases of papillary cancers, PTC; 8 follicular cancers, FTC; 12 medullary cancers, MTC and 11 anaplastic cancers, ATC), using ELISA assays. The control consisted of 27 healthy volunteers. Statistically significant lower concentrations of Ang-1 were found in patients with thyroid cancers as compared with the control ( p < 0.003). The levels of Ang-2 and Tie-2 did not differ significantly between thyroid cancer patients and control. We have also compared the results of Ang-1, Ang-2, and Tie-2 determinations obtained in different histopathological subgroups of cancer patients. These results revealed lower Ang-1 concentrations in ATC ( p < 0.05), MTC ( p < 0.02), FTC ( p < 0.01) and in PTC patients ( p < 0.05) than control. We have also observed lower Ang-2 concentration in PTC patients ( p < 0.03) and Tie-2 in FTC patients ( p < 0.02 ) in comparison to controls. In conclusion, the Angs/Tie-2 system dysfunction may play an important role in thyroid cancerogenesis and decreased concentration of Ang-1 in serum can be a useful additional biomarker for the presence of thyroid cancers.

Magnetic resonance-guided biopsy of suspicious breast lesions with a handheld vacuum biopsy device

To evaluate a handheld vacuum-assisted device system for magnetic resonance image (MRI)-guided breast lesion biopsy. In 32 patients, a total of 42 suspicious breast lesions (mean diameter 7.5 mm for mass lesions, 11.6 mm for non-masslike diffuse lesions) seen with MRI (no suspicious changes in breast ultrasound or mammography) were biopsied (27 lateral, 15 medial) using a 10G vacuum-assisted breast biopsy device under MR guidance. Histology of biopsy specimens was compared with final histology after surgery or follow-up in benign lesions. In all biopsies, technical success was achieved. Histology revealed 11 lesions with ductal carcinoma in situ (DCIS) or invasive cancer, three with intermediate lesions (LCIS) and 28 with benign breast lesions (adenosis, infected hematoma). In one patient with discordant results of MRI and histology, surgical excision revealed medullary cancer. In the follow-up (mean 18 months) of the histological benign lesions, no breast cancer development was observed. Besides minor complications (hematoma, n = 6), with no further therapeutic interventions, no complications occurred. MRI-guided breast lesion biopsy using a handheld vacuum-assisted device is a safe and effective method for the work-up of suspicious lesions seen with breast MRI without changes in mammography or ultrasound. In the case of discordant histology of vacuum biopsy and breast MRI appearance, surgical excision is recommended.

Management of Thyroid Cancer and Related Nodular Disease

Apart from the chapter on pathology, this is a single author book and has most of the advantages and few of the drawbacks of such an approach. The author is a nuclear medicine physician/‘thyroidologist' with several decades of recent practice at Stanford in USA, preceded by working in the UK and enriched by sabbaticals elsewhere. The target audience is wide, mostly within the medical specialties in a wide range of disciplines but also nurses, students and patients. The detail of the text and comprehensive scope of the reviews make it entirely suitable for those with a major interest in thyroid cancer who will appreciate it for its clear presentation of principles and rationale for treatments and as a guide to the management of difficult problems. An understanding of basic principles is not assumed. The sections on epidemiology and radiation physics and dosimetry especially develop the principles clearly and logically and are well worth attention. It therefore functions well both as a book to dip into for specific questions and as a comprehensive presentation of the evidence and process necessary to provide a modern practice. There is a consistent approach with a logical progression between the chapters that reinforce the principles without undue repetition. The broad personal experience provides a series of anecdotes and, while personal preferences are certainly expressed, these are always placed on an extensive and balanced background of literature review. The volume is some 398 A4 pages long and comprises 12 chapters. There are extensive references for each ranging from 60 or so for the epidemiology to 680 for the main management chapter. The majority relate to papillary and follicular tumours but lymphoma, medullary and anaplastic cancers are presented. The anatomy, physiology, aetiology and pathology are well covered with up to date information, especially on the genetics. There is good use of illustrative photographs and clear explanatory diagrams. The principles of nuclear medicine scanning, the interpretation of the scans and the physics underlying therapy are explained from first principles and would be very valuable to all non-nuclear medicine physicians in the field. He does not fail to tackle the current controversial issues that are the extent of surgery necessary, the need for ablation, the use and difficulties with thyroglobulin and the developing role of recombinant TSH. Very specific guidelines are outlined and examples of information sheets provided. These could be easily adapted for centres needing to update their own written information. I have few and minor reservations. In view of the detail in which the majority of the book is presented, a greater discussion about loco-regional relapse, its impact on the patient and its management and outcome would have been a welcome addition. It is generally very well presented but there are places where the figures are at some distance from the text that refers to them and impedes the ease of reading. In summary this is a well-presented text, comprehensive in content, logical and well presented. It combines clear management principles based on both personal experience and extensive familiarity with the literature. It is highly recommended for those with a major interest in thyroid cancer and would be a good reference volume on this topic for a library.

Hormonal therapy for menopause and breast-cancer risk by histological type: a cohort study and meta-analysis

Little information is available on how the risk of breast cancer associated with the use of hormone therapy for menopause varies by histological type. We aimed to describe such associations for eight histological types of breast cancer. Analyses are based on 1 031 224 postmenopausal women recruited in 1996-2001 into a nationwide UK cohort study, and followed for incident cancer and death. Relative risks associated with use of hormone therapy were estimated for eight histological types of breast cancer. During 3.6 million person-years of follow-up, 14 102 breast cancers were diagnosed, of which 13 782 (98%) had histological type recorded: 11 869 (86%) were invasive, including 8007 ductal, 1526 lobular, 365 mixed ductal-lobular, 492 tubular, 71 medullary, and 148 mucinous cancers; and 1913 (14%) were in situ, including 1443 ductal and 86 lobular cancers. The relative risks of invasive breast cancer in current users compared with never users of hormone therapy varied significantly according to tumour histology overall (p<0.0001), for users of oestrogen-only therapy (p=0.0001), and for users of oestrogen-progestagen therapy (p<0.0001). The largest relative risks in current compared with never users of hormone therapy were seen for lobular (relative risk 2.25, 95% CI 2.00-2.52), mixed ductal-lobular (2.13, 1.68-2.70), and tubular cancers (2.66, 2.16-3.28). The relative risks for ductal and mucinous cancers were 1.63 (95% CI 1.55-1.72) and 1.58 (1.08-2.31), respectively. The risk of medullary cancer was not increased (0.74, 0.43-1.28). The relative risk of in-situ disease in current users compared with never users of hormone therapy also varied significantly according to histological type (p=0.03), with a relative risk for lobular carcinoma in situ of 2.82 (1.72-4.63) and 1.56 (1.38-1.75) for ductal carcinoma in situ. The effects of hormone therapy on invasive ductal, lobular, and tubular cancer were generally greater for oestrogen-progestagen therapy than for oestrogen-only therapy, and were attenuated with increasing body-mass index (BMI). The risks associated with use of hormone therapy for menopause differ by histological type of breast cancer, and are substantially attenuated with increasing BMI.

Disparities between male and female patients with thyroid cancers: sex difference or gender divide?

This investigation was undertaken to quantify histopathological disparities between male and female patients with sporadic and hereditary thyroid cancers, which may reflect a biological 'sex difference' or a behavioural 'gender divide'. Retrospective cohort analysis (November 1994-January 2006). 1298 Consecutive surgical patients with sporadic papillary (n = 587), sporadic follicular (n = 232), sporadic medullary (n = 320), and hereditary medullary thyroid cancers (n = 159) from a tertiary referral centre. Age at diagnosis of cancer, primary tumour diameter, frequency of extrathyroidal extension, lymph node and distant metastases, and cancer subtypes. Primary diameters of sporadic tumour entities (papillary, 26.0 vs. 19.3 mm; follicular, 54.9 vs. 35.1 mm; and medullary, 27.9 vs. 20.8 mm), but not hereditary medullary cancers, were significantly (P <or= 0.001) larger in male patients. Likewise, lymph node metastases from sporadic papillary cancers (60%vs. 44%, P < 0.001), the insular subtype in sporadic follicular cancers (22%vs. 8%, P = 0.003), extrathyroidal extension of sporadic medullary cancers (35%vs. 15%, P < 0.001), and distant metastases from sporadic medullary cancers at the most recent operation (29%vs. 15%, P = 0.002) were seen significantly more often with male patients. No significant differences were observed between male and female patients with hereditary medullary cancers, half of which were detected by screening. Our data emphasize the need for earlier diagnosis and intervention in male patients to ensure that male sex will cease sometime to constitute an ominous prognostic marker of advanced thyroid cancer.

Role of 18F-FDG-PET and PET/CT imaging in thyroid cancer

In patients affected by differentiated thyroid cancer (DTC), the lacking of 131Iodine trapping by metastatic tissue does not allow 131Iodine whole body scintigraphy to visualize matastatic spread as well as the use of 131Iodine therapy to cure such metastatic spread. Prognosis of 131Iodine-negative DTC metastasis, so-called non-functioning metastasis, is significantly worst. In these patients an early diagnosis of non-functioning metastasis and their surgical extirpation remains the optimal therapeutic approach. In this view, a high sensitive localizing imaging different form 131Iodine whole body scintigraphy is required. Ultrasonography is characterized by a relatively high sensitivity in these patients but it is highly operator-dependent and, moreover, it can be used to explore neck alone. Computed tomography (CT) scan and magnetic resonance (MR) imaging are characterized by a relatively low sensitivity even if they are useful to provide the surgeon with anatomical information of the operating basin. Various tumor-seeking radiotracers have been proposed, mainly using SPECT as 201Thallium, 99mTc-Sestamibi and 99mTc-Tetrofosmin with good results. Even more favorable results have been reported with some positron radiotracers, mainly the 18F-FDG with PET and more recently with PET/CT tomographs. The typical indication to performing with examination is the DTC patient previously treated by total thyroidectomy and 131Iodine ablative therapy, with increased serum thyroglobulin (Tg) or anti-thyroglobulin (TgAb) antibodies during follow-up but with negative 131Iodine whole body scintigraphy even obtained after high, therapeutic 131Iodine doses. Several studies in literature have reported high sensitivity (up to 85%) and specificity (up to 95%) of FDG-PET in metastatic DTC patients. The integrated PET/CT fusion imaging systems, seem able to provide some additional advantages over PET alone, mainly related to a better anatomical localization of the hypermetabolic metastatic lesions. A change in the management of DTC patients affected by non-functioning metastatic spread not visualized by other imaging techniques has been reported in 30% of patients. Lastly, the role of PET and PET/CT fusion imaging systems seem to be promising also in patients affected by medullary thyroid carcinoma (MTC), especially for the detection of neck and mediastinal lesions, with a sensitivity superior to the other currently available imaging methods, however the data reported on medullary cancer are little and further studies are needed to elucidate the preliminary promising results.

Circulating chromogranin-a (CgA) as a useful marker in the diagnosis and follow up of neuroendocrine tumors (nets): An Italian multicenter observational study (cromAnet)

14091 The primary objective of this study was to validate the clinical significance of blood evaluation of CgA in NET patients at the diagnosis (PHASE I) and during 2 years of follow-up (PHASE II). From May 2003 to October 2004, 276 patients entered the study from 40 Italian centers: 270 were evaluable. All basal and every 3 months collected CgA blood samples were centrally measured in two reference laboratories (Orbassano-Turin and Venice) where ELISA (DAKO,Denmark) or IRMA (CIS-Schering, France) were performed to look at the correlation between the two methods and their sensitivity and specificity. Lab results at the baseline have been recently published (Leon et al., Intern. J. Biol. Markers, 2005). We are now collecting all the correlations between CgA and type and place of NETs; tumor bulk; metastatization; presence or not of specific syndrome;proliferation activity (Ki67); octreoscan; tumor specific markers. 223 patients (83%) had gastroenteropancreatic tumors, whereas 24 (9%) had medullary tyroid cancer, 16 (6%) Merkel cell carcinoma and 6 parathyroid NETs, pheochromocytoma, paraganglioma. Only 26% of GEP tumors presented with specific symptoms. At the entry in the study 58% of patients had a new diagnosis, 23% were in stable disease, whereas 18% had metastatic disease. According to the recent W.H.O. histologic classification (Solcia et al, 2000), 36% specific symptomatic patients had NE tumor, 57% well differentiated cancer and only 3% poor differentiated cancer, whereas 31% not symptomatic patients had NE tumor, 48% well differentiated and 16% poor differentiated cancer. This is the largest study worldwide performed on this topic and all the data about the correlation among all patient variables and CgA blood values will be ready in April 2006. Follow-up data will be evaluable next year. [Table: see text]

Identification of genes with altered expression in medullary breast cancer vs. ductal breast cancer and normal breast epithelia

Medullary breast cancer (MCB) is a morphologically and biologically distinct subtype that, despite cytologically highly malignant characteristics, has a favorable prognosis compared to the more common infiltrating ductal breast carcinoma. MCB metastasizes less frequently, which has been attributed to both immunological and endogenous cellular factors, although little is known about the distinct biology of MCB that may contribute to the improved outcome of MCB patients. To identify candidate genes, we performed gene array expression analysis of cell lines of MCB, ductal breast cancer and normal breast epithelia, and the differential expression of a panel of candidate genes was further validated by quantitative PCR and immunohistochemical analysis of cell lines and tumor biopsies. A limited number of genes, including several members of the GAGE and insulin growth factor binding protein (IGFBP) gene families, Vav1, monoglyceride lipase and NADP+-dependent malic enzyme, exhibited altered expression in MCB vs. ductal breast cancer, and the differences for some of these genes were confirmed on an extended panel of cell lines by quantitative PCR. Immunohistochemical analysis further established that the expression of monoglyceride lipase was restricted to ductal breast cancer and present in 77% of these tumors, while Vav1 was restricted to MCB and present in 60% of tumors. In this study, we have identified genes that are differentially expressed in MCB vs. ductal breast cancer and further analysis of the gene products should illuminate the biological differences between MCB and ductal breast cancer.

Comment interpréter une hypercalcitoninémie ?

Propose Today, calcitonin assay is used for the diagnosis of thyroid medullary cancer in the context of nodular thyroid disease. Calcitonin is an excellent marker of thyroid medullary cancer but some hypercalcitoninemia can also be related to other diseases, such as renal failure, endocrine tumors other than thyroid medullary cancer and sometimes to C cell hyperplasia, which is a not well-defined situation. Recent studies contributed to define calcitoninemia thresholds, which guide decision and avoid excessive invasive treatment. Current knowledge and key points After a brief reminder of physiological role of calcitonin and assays, the difficulties encountered in interpreting hypercalcitoninemia and its potential causes other than thyroid medullary cancer are addressed. Recent studies, on large series, now allow a better knowledge of specificity and sensitivity of calcitonin measurement in patients with nodular thyroid disease and a well-argued management. Future prospects and projects In the future, calcitonin dosage will be ordered even more frequently, as some authors recommend it for the diagnosis of thyroid nodule. It is up to us to know how to use this remarkable marker, by considering all possible situations of benign hypercalcitoninemia and reserving aggressive treatments for patients who really need them.

Current concepts of 131I therapy in oncology: Indications, methods and follow up

Radioiodine in various forms (as sodium iodide and as the iodinated compounds MIBG, LIPIODOL, et al.) has been used as a therapeutic agent in oncology. Differentiated thyroid carcinoma (DTC) has been successfully treated by 131I therapy. Neuroendocrine tumors can be treated by palliative therapy, including Meta 131Iodobenzylguanidine therapy (131I-MIBG). Diagnostic 131I or 123I-MIBG scintigraphy is usually performed to image neuroblastoma and malignant pheochromocytoma. Following the establishment of the diagnosis, 131I-MIBG may be applied as a therapeutic agent but with limited success. Hepatocellular carcinoma (HCC) is treated by surgery only in 10% of patients. In others, palliative therapy should be administered. Radionuclide therapy for this disease is a therapeutic option with a major advantage compared to systemic chemotherapy, estrogen and progesterone therapy, and immunotherapy. 131I-lipiodol can be used to treat HCC without side effects. Compared to untreated patients, those who received 131Ilipiodol, showed significantly better survival and a decreased recurrence rate. The modern aspect of the neoplasm treatment involves radioimmunotherapy with radioiodine and some other radionuclides. Monoclonal antibody therapy with radioiodine has been extensively succeeded in the therapy of B-cell non-Hodgkin?s lymphoma, prostate cancer. Radioimmunotherapy is also efficiently performed in some other malignancies such as: medullary thyroid carcinoma, breast cancer, colorectal cancer and malignant brain tumors. Radioimmunotherapy will play a key role in the treatment of malignant diseases, especially hematopoietic neoplasms during this millennium.

Cyclin E Overexpression Obstructs Infiltrative Behavior in Breast Cancer: A Novel Role Reflected in the Growth Pattern of Medullary Breast Cancers

Cell cycle deregulation is a prerequisite in tumor development and overexpression of cyclin E, a major G1-S regulator, is often observed in breast cancer and is further linked to poor prognosis. By overexpressing cyclin E in a retinoblastoma-inactivated breast cancer cell line, we induced significant alterations in the expression of genes associated with proliferation and cell adhesion. Rearrangements of the actin cytoskeleton in addition to increased adhesive properties, decreased motility, and invasive potential in functional assays, indicated an overall abrogated mobility. Consistent in vivo findings were obtained upon investigation of 985 primary breast cancers, where cyclin E-high tumors predominantly (67%) displayed a low infiltrative, pushing growth pattern. Furthermore, medullary breast cancers, a subtype defined by its pushing, delimited growth, exhibited a remarkable frequency of cyclin E deregulation (87%) compared with other histologic subtypes (5-20%). Taken together, our results suggest the novel role of cyclin E in modeling infiltrative behavior. The consequences of cyclin E overexpression in breast cancer seems to be multiple, including effects on proliferation as well as growth patterns, a scenario that is indeed observed in the archetype of cyclin E-overexpressing medullary breast cancers.

Establishment of the hu-PBL-SCID Mouse Model for the Investigation of Thyroid Cancer

New experimental models of human neoplastic diseases attempt to mimic the human environment that fostered the development of disease in cancer patients. The aim of the present study was to establish a human lymphocyte-engrafted, severe combined immunodeficient (hu-PBL-SCID) mouse model to investigate thyroid cancer and to evaluate the potential use of this model for cancer immunotherapy. Thyroid neoplastic tissues were obtained from ten patients (one follicular adenoma, five papillary, one follicular, one anaplastic and two medullary cancers). One 8 x 4 x 3 millimeter sample from each tumor was cut into two pieces of identical size and transplanted into two SCID mice. In each case, one of the two mice was injected intraperitoneally with lymphocytes from the same tumor patient for the reconstitution of the human immune system (Group A), while the other animal received no lymphocytes (Group B). The engraftment of the tumors was successful in all cases. The growth rate was highly dependent on the histological type. When histologies were compared before implantation and after the removal of the implants, the characters of the tumors proved to be unchanged, except one case where an anaplastic cancer arose from a papillary tumor. Macrophages were present in all but one papillary cancer. All differentiated thyroid cancers were infiltrated by T and B lymphocytes. Lymphocytes and macrophages disappeared from 19/20 grafts by week 16. However, in one case from group A lymphocytes were detected four months after the transplantation. In another case from group A, one papillary cancer spontaneously decreased in size and disappeared. Before implantation, HLA-DR expression was detected in every papillary cancer. HLA-DR expression in the grafts was not seen in 3/5 cases by week 16. In conclusion, an animal model has been established for the investigation of human thyroid cancer, by which the analysis of anti-tumor immunity, as a postulate of immune therapy, may be possible.

Medullary and papillary carcinoma of the thyroid gland occurring as a collision tumour: report of three cases with molecular analysis and review of the literature

We report the simultaneous occurrence of medullary thyroid carcinoma (MTC) and papillary thyroid carcinoma (PTC), presenting as spatially distinct and well-defined tumour components, in three cases. In the first patient, histology, immunohistochemistry and electron microscopy demonstrated an MTC in the one nodule and PTC in two additional lesions. Non-neoplastic thyroid parenchyma separated the three nodules. Metastasis from PTC was diagnosed in a regional lymph node. Genetic analysis of both tumour components showed a distinctive mutational pattern: in the MTC a Cys634Arg substitution in exon 11 of the RET gene and in the two PTC foci a Val600Glu substitution in exon 15 of the BRAF gene. The other two patients are members of a large multigenerational family affected with familial MTC due to a germline mutation of the RET gene (Ala891Ser). Both patients harboured, besides medullary cancer and C-cell hyperplasia, distinct foci of papillary thyroid cancer, which was positive for Val600Glu BRAF mutation. Review of the literature disclosed 18 similar lesions reported and allowed the identification of different patterns of clinical presentation and biological behaviour. So far, the pathogenesis of these peculiar cases of thyroid malignancy has been completely unknown, but an underlying common genetic drive has been hypothesised. This is the first report in which two mutations, in the RET and BRAF genes, have been identified in three cases of MTC/PTC collision tumour, thus documenting the different genetic origin of these two coexisting carcinomas.

ERβ in normal and malignant breast

Both ERα and ERβ are expressed in not only normal breast of the rodent, cow, monkey and human, but also in breast cancer. Cells that express ERα are found within the luminal epithelium, but not in the myoepithelium or stroma in the human breast. ERβ, on the other hand, is expressed not only in the luminal epithelial cells, but also in myoepithelial cells, stromal cells and in passenger lymphocytes. This widespread distribution of ERβ suggests multiple roles for ERβ in the mammary gland. We have shown that in the rodent mammary gland ERβ is the dominant ER, and that, in response to E2, ERα but not ERβ is downregulated in the early G1 phase of the cell cycle. Cells that contain ERα receive the signal to proliferate from E2, and within 4 hours of that signal ERα is lost from the nucleus. The cells then go through a complete cycle and ERα reappears in daughter cells. ERβ levels do not change in cell nuclei during the cell cycle. This pattern of ER regulation holds true in human breast cancer since ERα is never co-localized with proliferation markers in breast cancer samples. This means that under the conditions of a constant high level of E2, ERα does not reappear in the nucleus. A similar situation exists during pregnancy when there is a constant high level of E2 and there is no ERα in the mammary epithelium. This resistance to the proliferative response to E2 in the presence of a constant high dose of E2 probably explains the very successful use of high-dose E2 in the treatment of breast cancer. ERβ, on the other hand, appears to have a differentiative role not a proliferative role in the mammary gland, and the lactating rodent mammary gland of ERβ-/- mice does not express gap junction and adhesion proteins, typical indicators of fully differentiated cells. In recent years there have been several publications showing that ERβ is expressed in human breast cancer, and conclusions and speculations about a causative role for ERβ in breast cancer development and/or progression have been made. We have studied 500 frozen breast biopsies in collaboration with Prof. RC Coombes, London, in order to clarify the role of ERβ in normal and malignant breast. In this study we measured ERα and ERβ proteins by several techniques (immunohistochemistry, western blotting, ligand binding in sucrose gradients, and RT-PCR) in various human samples obtained from both benign breast and malignant breast. We found that ERβ is the predominant estrogen receptor in the normal mammary gland and in benign breast disease. There is very little ERα in the normal mammary gland. This low expression of ERα is one of the striking differences between rodents and humans. This is in stark contrast to ERβ, which is expressed in 80% of epithelial cells and is also present in the stroma. We found that ERα is abundantly expressed in invasive and in situ ductal carcinoma but not in medullary cancer. ERβ is also expressed in breast cancer, both ductal and medullary. In this study we also found that, in the human breast, the major ER in breast stroma is ERβ. This surprising finding has necessitated several new lines of investigation about the function of ERβ in the breast. It has long been thought that ERα in the stroma was responsible for secretion of growth factors in response to E2 and that these growth factors were responsible for epithelial cell proliferation. The discovery that it is ERβ that is present in the stroma might suggest a role of ERβ in growth factor secretion.

Dual Inhibition of RET and FGFR4 Restrains Medullary Thyroid Cancer Cell Growth

Abstract Medullary thyroid cancer is frequently an aggressive form of carcinoma for which there are currently no effective forms of systemic therapy. These carcinomas arise as a result of activating mutations in the RET proto-oncogene transmembrane tyrosine kinase receptor. We, therefore, examined the potential efficacy of the tyrosine kinase inhibitor STI571 on the growth of human TT medullary cancer cells in vitro and in xenografted severe combined immunodeficiency mice. Treatment with STI571 resulted in inhibition of RET phosphorylation, cell proliferation, tumor growth and invasiveness. Based on the profile of expression of fibroblast growth factor receptors (FGFR), we examined the effects of FGFR tyrosine kinase inhibition using the small molecule FGFR inhibitor PD173074. This inhibitor resulted in abrogation of fibroblast growth factor-1-mediated FGFR4 phosphorylation in TT cells, an effect that was accompanied by significant arrest of cell proliferation and tumor growth in vivo. Moreover, the combination of STI571 and PD173074 resulted in greater suppression of cell proliferation in vitro and tumor control in vivo than that achieved with either agent alone. These data highlight RET and FGFR4 as therapeutic targets and suggest a potential role for the combined use of tyrosine kinase inhibitors in the management of inoperable medullary thyroid cancers.

A population-based assessment of the clustering of breast cancer in families eligible for testing of BRCA1 and BRCA2 mutations

BackgroundThe prevalence of families eligible for BRCA1/2 mutation testing in the population burden of breast cancer was analysed and the aggregation of breast cancer in these families was explored. Patients and methodsThe families of the Swedish Family-Cancer Database with at least three generations (N=944723) were classified according to the criteria proposed by the German Consortium for Hereditary Breast and Ovarian Cancer for BRCA1/2 mutation testing. We calculated the proportion of women with breast cancer in the classified families and used standardised incidence ratios (SIRs) to estimate the risk of histology specific breast cancers in families with suspected BRCA1/2 mutations. ResultsFamilies with two breast cancers before the age of 50 years included 1.8% of the breast cancer patients; 1% of the women with breast cancer belonged to families with breast and ovarian cancers. The SIR of female breast cancer was lowest in families with male breast cancer and highest in families with two women affected by breast cancer under the age of 50 years. The SIRs of medullary breast cancer agreed with the BRCA1 mutation prevalences detected by the German Consortium for Hereditary Breast and Ovarian Cancer. ConclusionsMost of the breast malignancies in families with male breast cancer are likely to be related to BRCA2 mutations. Non-BRCA1/2 related effects are probably involved in the strong clustering of breast cancer in families with early onset breast and ovarian cancers.

Typical medullary breast carcinomas have a basal/myoepithelial phenotype

Medullary breast cancer (MBC) is a rare, diagnostically difficult, pathological subtype. Despite being high grade, it has a good prognosis. MBC patients have an excess of BRCA1 germ-line mutation and reliable identification of MBC could help to identify patients at risk of carrying germline BRCA1 mutations or in whom chemotherapy could be avoided. The aim of this study was therefore to improve diagnosis by establishing an MBC protein expression profile using immunohistochemistry (IHC) on tissue-microarrays (TMA). Using a series of 779 breast carcinomas ('EC' set), diagnosed initially as MBC, a double-reading session was carried out by several pathologists on all of the histological material to establish the diagnosis as firmly as possible using a 'medullary score'. Only MBCs with high scores, i.e. typical MBC (TMBC) (n=44) and non-TMBC grade III with no or low scores (n=160), were included in the IHC study. To validate the results obtained on this first set, a control series of TMBC (n=17) and non-MBC grade III cases (n=140) ('IPC' set) was studied. The expression of 18 proteins was studied in the 61 TMBCs and 300 grade III cases from the two sets. The global intra-observer concordance of the first reading for the diagnosis of TMBC was 94%, with almost perfect kappa (kappa) of 0.815. TMBC was characterized by a high degree of basal/myoepithelial differentiation. In multivariate analysis with logistic regression, TMBC was defined by the association of P-cadherin (R=2.29), MIB1 > 50 (R=3.80), ERBB2 negativity (R=2.24) and p53 positivity (RR=1.45).

Prognostic Relevance of Gene Amplifications and Coamplifications in Breast Cancer

Multiple different oncogenes have been described previously to be amplified in breast cancer including HER2, EGFR, MYC, CCND1, and MDM2. Gene amplification results in oncogene overexpression but may also serve as an indicator of genomic instability. As such, presence of one or several gene amplifications may have prognostic significance. To assess the prognostic importance of amplifications and coamplifications of HER2, EGFR, MYC, CCND1, and MDM2 in breast cancer, we analyzed a breast cancer tissue microarray containing samples from 2197 cancers with follow-up information. Fluorescence in situ hybridizations revealed amplifications of CCND1 in 20.1%, HER2 in 17.3%, MDM2 in 5.7%, MYC in 5.3%, and EGFR in 0.8% of the tumors. All gene amplifications were significantly associated with high grade. HER2 (P < 0.001) and MYC amplification (P < 0.001) were also linked to shortened survival. In case of HER2, this was independent of grade, pT, and pN categories. MYC amplification was almost 3 times more frequent in medullary cancer (15.9%), than in the histologic subtype with the second highest frequency (ductal; 5.6%; P = 0.0046). HER2 and MYC amplification were associated with estrogen receptor/progesterone receptor negativity (P < 0.001) whereas CCND1 amplification was linked to estrogen receptor/progesterone receptor positivity (P < 0.001). Coamplifications were more prevalent than expected based on the individual frequencies. Coamplifications of one or several other oncogenes occurred in 29.6% of CCND1, 43% of HER2, 55.7% of MDM2, 65% of MYC, and 72.8% of EGFR-amplified cancers. HER2/MYC-coamplified cancers had a worse prognosis than tumors with only one of these amplifications. Furthermore, a gradual decrease of survival was observed with increasing number of amplifications. In conclusion, these data support a major prognostic impact of genomic instability as determined by a broad gene amplification survey in breast cancer.

Angiogenic Profile of Breast Carcinoma Determines Leukocyte Infiltration

To study the relationship between the angiogenic profile and leukocyte infiltration of tumors, single cell suspensions of archival frozen medullary and ductal breast cancer tissues were analyzed by flow cytometry. The amount of leukocytes and endothelial cells was measured, as well as the expression of intercellular adhesion molecule-1 (ICAM-1) on the endothelial cell fraction. A significantly higher number (3.2-fold) of infiltrating leukocytes was observed in medullary carcinoma. The composition of this infiltrate was similar to that seen in ductal carcinomas. The more intense infiltrate was explained by the approximately 3-fold enhanced endothelial ICAM-1 expression in medullary carcinoma. The angiogenic profile of all tumors was assessed by quantitative real-time reverse transcription-PCR analysis. Vascular endothelial growth factor (VEGF)-C and VEGF-D, but not VEGF-A, basic fibroblast growth factor, placental growth factor, and angiopoietins 1, 2, and 3 showed a relatively higher level of expression in ductal carcinoma than in medullary carcinoma. In vitro, both VEGF-C and VEGF-D were found to decrease endothelial ICAM-1 expression in the presence of basic fibroblast growth factor. These data suggest that in vivo angiogenic stimuli prevent the formation of an effective leukocyte infiltrate in tumors by suppressing endothelial ICAM-1 expression.

Gastric medullary carcinoma, a distinct entity associated with microsatellite instability‐H, prominent intraepithelial lymphocytes and improved prognosis

To determine the clinicopathological and molecular features of gastric medullary cancer. Clinicopathological review and microsatellite instability (MSI) analysis were carried out on 17 gastric medullary and 64 non-medullary cancers. In addition to characteristic histopathology, gastric medullary cancers had certain prominent features: (i) the average survival time was longer in medullary and low-grade non-medullary cancers than in high-grade (P = 0.004); (ii) serosal involvement was less common in medullary cancers (29.4%, 5/17) than in non-medullary cancers (9.4%, 6/64) (P < 0.05) while pushing borders were more common in medullary cancers (70.6%, 12/17 versus 17.2%, 11/64, P = 0); (iii) the presence of intraepithelial lymphocytes (IELs) in medullary and non-medullary cancers was 2380/10 high-power field (HPF) and 147/10 HPF (P = 0), respectively. Both peritumoural infiltrating lymphocytes (pTIL) and a Crohn's-like reaction were more common in medullary cancers than in non-medullary (pTIL 35.3%, 6/17 versus 3.1%, 2/64; a Crohn's-like reaction 70.6%, 12/17 versus 32.8%, 21/64; P < 0.05); (iv) medullary and high-grade non-medullary cancers were more associated with reduced ECD expression in comparison with low-grade cancers (P < 0.05); (v) higher MSI-H (Bat26+) rate was observed in medullary cancers (41.2%, 7/17) than in non-medullary (1.6%, 1/64) (P = 0). Gastric medullary cancer has distinct clinicopathological features and genetic alterations. Two subtypes of gastric medullary cancers, Bat26+ and Bat26-, might have prognostic implications, thus analysis of Bat26 may be of clinical value.