Lola Anagnostaki

Tissue response to silicone tubes used to repair human median and ulnar nerves

Silicone tubes of appropriate sizes were used to enclose the injured zone of transsected ulnar and median nerves in the human forearm as an alternative to conventional microsurgical repair of the nerve trunk. A gap measuring 3-5 mm was left intentionally between the nerve ends inside the tube. The clinical early results from a prospective randomised study that compared these two principles have recently been presented. Seven patients (five men and two women), aged 15-49 years (median 20) were reexplored 12-44 months (median 22) after the initial procedure because of local discomfort from the tube in four patients. There was a new nerve structure bridging the former gap and in most cases it was impossible to distinguish the site of the injury. In all cases there was a thin capsule around the silicone tube that microscopically consisted of connective tissue with thin walls and no signs of inflammation, granuloma or macrophages (n = 4), while in two cases a mild foreign body reaction was seen at a single site (n = 1) or at patchy areas (n = 1). These results indicate that after more than one year there is a limited tissue reaction around silicone tubes used to repair median and ulnar nerves in humans.Silicone tubes of appropriate sizes were used to enclose the injured zone of transsected ulnar and median nerves in the human forearm as an alternative to conventional microsurgical repair of the nerve trunk. A gap measuring 3-5 mm was left intentionally between the nerve ends inside the tube. The clinical early results from a prospective randomised study that compared these two principles have recently been presented. Seven patients (five men and two women), aged 15-49 years (median 20) were reexplored 12-44 months (median 22) after the initial procedure because of local discomfort from the tube in four patients. There was a new nerve structure bridging the former gap and in most cases it was impossible to distinguish the site of the injury. In all cases there was a thin capsule around the silicone tube that microscopically consisted of connective tissue with thin walls and no signs of inflammation, granuloma or macrophages (n = 4), while in two cases a mild foreign body reaction was seen at a single site (n = 1) or at patchy areas (n = 1). These results indicate that after more than one year there is a limited tissue reaction around silicone tubes used to repair median and ulnar nerves in humans.

Oestrogen receptors alpha and beta show different associations to clinicopathological parameters and their co-expression might predict a better response to endocrine treatment in breast cancer.

Aims: The majority of all breast cancers are hormone responsive, traditionally defined by the expression of oestrogen receptor (ER) α and/or progesterone receptors. In contrast to ERα, the clinical significance of the relatively recently identified ERβ is still unclear. This study aimed to define the relationship between ERβ and clinicopathological parameters in a mixed cohort of breast cancer and, furthermore, to investigate the impact of ERβ expression on disease outcome. Methods: The immunohistochemical expression of ERα and ERβ was analysed in tissue microarrays containing a total number of 512 tumours with all incident breast cancers diagnosed at the Malmö University Hospital between 1988 and 1992. Results: 78% of the tumours were ERα positive and 50% were ERβ positive. ERβ correlated positively with ERα (p = 0.001). In contrast to ERα, ERβ was not associated with any important clinicopathological variables. Furthermore, no overall prognostic significance could be demonstrated for ERβ. In the ERα-positive subgroup, however, a low expression of ERβ correlated with a decreased disease-free survival in patients receiving endocrine treatment (p = 0.003). Conclusions: Although interrelated, ERα and ERβ seem to be differentially associated to clinicopathological parameters, and this would support the fact that they might have different functions in vivo. Furthermore, ERβ might be a predictive marker of response to endocrine therapy, although this needs to be confirmed in additional studies, preferably randomised trials.

Protein expression and cellular localization in two prognostic subgroups of diffuse large B-cell lymphoma: Higher expression of ZAP70 and PKC-beta II in the non-germinal center group and poor survival in patients deficient in nuclear PTEN

Patients diagnosed with diffuse large B-cell lymphoma (DLBCL) show varying responses to conventional therapy, and this might be contributed to the differentiation stage of the tumor B-cells. The aim of the current study was to evaluate a panel of kinases (ZAP70, PKC-β I and II and phosphorylated PKB/Akt) and phosphatases (PTEN, SHP1 and SHP2) known to be frequently deregulated in lymphoid malignancies. De novo DLBCL cases were divided into two subgroups, the germinal center (GC) group (14/28) and the non-germinal center (non-GC) or activated B-cell (ABC) group (14/28). ZAP70 and PKC-β II were expressed in a significantly higher percentage of tumor cells in the clinically more aggressive non-GC group compared with the prognostically favourable GC group. Also, the subcellular localization of PKC-β I and II differed in DLBCL cells, with the PKC-β I isoform being expressed in both the cytoplasm and nucleus, while PKC-β II was found exclusively in the cytoplasm. Loss of nuclear PTEN correlated with poor survival in cases from both subgroups. In addition, five cell lines of DLBCL origin were analyzed for protein expression and for mRNA levels of PTEN and SHP1. For the first time, we show that ZAP70 is expressed in a higher percentage of tumor cells in the aggressive non-GC subgroup of DLBCL and that PKC-β I and II are differently distributed in the two prognostic subgroups of de novo DLBCL.

HMG-CoA reductase expression in breast cancer is associated with a less aggressive phenotype and influenced by anthropometric factors

Although several studies have reported on the anti‐tumoural properties exerted by 3‐hydroxy‐3‐methylglutaryl‐coenzyme A reductase (HMG‐CoAR) inhibitors (statins), the in vivo expression of HMG‐CoAR in human cancer has been considerably less investigated. In our study, we examined the immunohistochemical expression of HMG‐CoAR in 511 incident breast cancers within the Malmö Diet and Cancer Study in order to explore its relationship to established clinicopathological and tumour biological parameters. Furthermore, the potential influence of estrogen exposure on HMG‐CoAR expression was assessed by performing Coxs proportional hazards analyses of the relationship between the use of hormone replacement therapy (HRT), obesity (waist circumference) and tumour‐cell specific HMG‐CoAR expression. We found that HMG‐CoAR was present in various fractions and intensities in the cytoplasm, sometimes with a membranous pattern, but not in the tumour cell nuclei. The expression of HMG‐CoAR was associated with a smaller tumour size (p = 0.02), low histological grade (p = 0.001), low Ki67 index (p = 0.004), ERα+ (p = 0.02), ERβ+ (p = 0.005), and high p27 expression (p = <0.001). The incidence of tumours with a high HMG‐CoAR‐expression was increased among HRT‐users, although this was not statistically significant in a heterogeneity analysis. Obesity was significantly associated with a high HMG‐CoAR expression assessed both as a high (>50%) fraction of positive cells (relative risk: 2.06; 95% confidence interval: 1.20–3.51), and a strong staining intensity (2.33: 1.08–5.02). In summary, we demonstrate that HMG‐CoAR is differentially expressed in breast cancer and that a high expression is associated with prognostically favourable tumour parameters. Moreover, estrogen related life‐style and anthropometric factors might indeed regulate HMG‐CoAR expression.

Pathology parameters and adjuvant tamoxifen response in a randomised premenopausal breast cancer trial.

Background: Subgroups of breast cancer that have an impaired response to endocrine treatment, despite hormone receptor positivity, are still poorly defined. Breast cancer can be subdivided according to standard pathological parameters including histological type, grade, and assessment of proliferation. These parameters are the net result of combinations of genetic alterations effecting tumour behaviour and could potentially reflect subtypes that respond differently to endocrine treatment. Aims: To investigate the usefulness of these parameters as predictors of the response to tamoxifen in premenopausal women with breast cancer. Materials/methods: Clinically established pathological parameters were assessed and related to the tamoxifen response in 500 available tumour specimens from 564 premenopausal patients with breast cancer randomised to either two years of tamoxifen or no treatment with 14 years of follow up. Proliferation was further evaluated by immunohistochemical Ki-67 expression. Results: Oestrogen receptor positive ductal carcinomas responded as expected to tamoxifen, whereas the difference in recurrence free survival between control and tamoxifen treated patients was less apparent in the relatively few lobular carcinomas. For histological grade, there was no obvious difference in treatment response between the groups. The relation between proliferation and tamoxifen response seemed to be more complex, with a clear response in tumours with high and low proliferation, whereas tumours with intermediate proliferation defined by Ki-67 responded more poorly. Conclusions: Clinically established pathology parameters seem to mirror the endocrine treatment response and could potentially be valuable in future treatment decisions for patients with breast cancer.

Breast tumours following combined hormone replacement therapy express favourable prognostic factors

The aim of the present study was to evaluate the association between different types of hormone replacement therapy (HRT) and risk of specific breast cancer subgroups. A population‐based prospective cohort study including 12,583 peri‐ or postmenopausal women were followed using record‐linkage with national cancer registries. During an average follow‐up of 4.5 years, 332 cases of invasive breast cancer were diagnosed. Tumour samples were available from 283 cases. These tumours were re‐evaluated according to histological type, grade, and mitotic index. Evaluation of tumours included estrogen and progesterone receptor status (ERα, ERβ and PgR), as well as expression of Ki67, HER2, cyclin D1 and p27. The incidence of breast cancer in current users of combined HRT (CHRT) was significantly higher than in non‐users. The difference corresponded to an adjusted relative risk (95% confidence interval) of 3.01 (2.35–3.84) as obtained using a Coxs proportional hazards analysis. CHRT was associated with lobular tumours (3.48:1.99–6.10), grade 1 tumours (4.46:2.79–7.13) and tumours with a low mitotic index (4.35:2.99–6.34). CHRT was not related to any specific subgroup in terms of ERα‐, ERβ‐ or PgR‐expression. CHRT was associated with low proliferating tumours, defined by the Ki67 index (3.58:2.60–4.93), HER2 amplified tumours (4.40:1.93–10.06), low expression of the oncogene cyclin D1 (3.14:2.32–4.23) and high expression of the tumour suppressor gene p27 (3.47:2.40–5.01). Use of estrogen‐alone HRT (ERT) was not associated with any statistically significant risk of breast cancer. We conclude that the use of CHRT is associated with an increased incidence of breast tumours with comparatively favourable prognostic factors.

Tissue microarray analyses of G1/S-regulatory proteins in ductal carcinoma in situ of the breast indicate that low cyclin D1 is associated with local recurrence

Ductal carcinoma in situ (DCIS) of the breast constitutes about 10% of all diagnosed breast cancers and, despite surgical removal, it may recur, either as DCIS or invasive breast cancer. Nuclear grade and growth pattern according to Andersen et al as well as surgical margins are factors that have been used to predict local recurrence, but ideally a set of tumour-specific factors should be identified and used as prognostic markers. Many cell cycle regulatory gene products have been shown to be involved in the formation of tumours and are either oncogenes or suppressor genes and involved in key processes in the transformation. We therefore characterised the cell cycle regulators cyclin E, cyclin D1, p27 and p16 in a material of DCIS cases arranged in a tissue microarray. With a manual tissue arrayer, 52% of the initial 177 DCIS samples were successfully targeted allowing immunohistochemical analyses of all four proteins in 92 cases of DCIS. As also observed in invasive breast cancer, there was a trend indicating that DCIS cases with high cyclin D1 were cyclin E low and oestrogen receptor-positive, whereas cyclin E high DCIS cases were cyclin D1 low and oestrogen receptor-negative. For the 64 patients that did not receive postoperative radiotherapy, there were 16 local recurrences (eight DCIS and eight invasive breast cancer) during a mean follow-up time of 63 months. Cyclin E, p27 or p16 were not associated with local recurrence, but interestingly cyclin D1 was significantly and inversely associated with local recurrence, both using univariate and multivariate analyses. In summary, using a tissue array approach we have shown that cyclin D1, besides growth pattern, is a prognostic marker for local recurrence in DCIS.

Anthropometric factors in relation to different tumor biological subgroups of postmenopausal breast cancer.

Overweight and obesity is associated with an increased risk of postmenopausal breast cancer. However, less is known about the impact of anthropometric factors on tumor pathology and biology. A Swedish population‐based prospective cohort study of 9,685 postmenopausal women not using hormonal replacement therapy (HRT) were followed for an average of 10.3 years during which 305 incident breast cancer cases were diagnosed. Invasive and sufficient tumor material was available in 248 cases. Pathological reevaluation of histological type and grade was conducted. Using a tissue microarray (TMA), the tumor expression of Ki67, HER2, ERα, ERβ, PgR, cyclin D1 and p27 was evaluated. Six anthropometric factors: height, weight, body mass index (BMI), waist‐ and hip circumference and body fat percentage were categorized by quartiles of baseline anthropometric measurements, and relative risks were calculated using multivariate Cox regression models. Invasive breast cancer incidence was increased for women in the higher quartiles of all anthropometric measurements. Height was positively associated with Grade I and ERα‐positive tumors. Women in the highest quartiles of weight, BMI, waist‐ and hip circumference and body fat percentage were all associated with tumors of ductal type, Grade II, low Ki67 index, HER2 negativity and low expression of the oncogene cyclin D1. Obesity was further associated with tumors expressing ERα and PgR but interestingly not ERβ. This study confirmed previously described associations between overweight/obesity and increased risk of postmenopausal breast cancer. Furthermore, obesity was associated with tumors expressing several markers corresponding with low malignancy.

Parity and age at first childbirth in relation to the risk of different breast cancer subgroups

The aim of the present study was to examine parity and age at first childbirth, in relation to the risk of specific breast cancer subgroups. A prospective cohort, The Malmö Diet and Cancer Study, including 17,035 women were followed with linkage to Swedish Cancer Registry until December 31, 2004. A total of 622 incident breast cancers were diagnosed during follow‐up and were evaluated regarding invasiveness, tumour size, axillary lymph node status, Nottingham grade, tumour proliferation (Ki67), HER2, cyclin D1 and p27. The tumours were also examined for WHO type and hormone receptor status. Nulliparity was associated with an overall increased risk of breast cancer, although not statistically significant (the relative risk was 1.39 with a 95% confidence interval of 0.92–2.08). Nulliparity was also associated with large tumours (>20 mm) (1.89: 0.91–3.91), high Ki67 levels (1.95: 0.93–4.10), high cyclin D1 levels (2.15: 0.88–5.27), grade III (2.93: 1.29–6.64) and HER2 positive tumours (3.24: 1.02–10.25). High parity was not statistically significantly associated with any specific breast cancer subgroup. Older age at first childbirth (>30) was associated with a slightly increased risk of breast cancer (1.39: 0.94–2.07). There was a statistically significant association between late first childbirth and lobular type (2.51: 1.01–6.28), grade III tumours (2.67: 1.19–6.02), high levels of cyclin D1 (2.69: 1.18–6.12) and low levels of p27 (2.23: 1.15–4.35). We conclude that nulliparity and late first childbirth are associated with relatively more aggressive breast cancer subgroups.

Expression of cyclin A1 and cell cycle proteins in hematopoietic cells and acute myeloid leukemia and links to patient outcome.

Abstract:  Abnormal expression of several key regulators essential for G1/S transitions has been implicated in tumorigenesis. A critical role of cyclin A1 in the development of acute myeloid leukemia (AML) has previously been demonstrated in transgenic mice. Our present study focused on the expression and prognostic significance of cyclin A1 and a panel of cell cycle regulatory proteins including cyclin A2, cyclin B1, cyclin E, CDK1, CDK2, p21 and p27 in bone marrow samples from 40 patients with AML. Freshly isolated CD34+ hematopoietic cells and bone marrow samples from 10 healthy donors were also assessed for cell type‐ and subcellular‐specific expression of the cell cycle regulatory proteins. The level of cyclin A1 expression was the only factor that showed a significant correlation with patient outcome. In log‐rank test stratified by levels of cyclin A1 expression, patients with high levels of cyclin A1 had significantly worse overall survival (OS) (P = 0.012) compared to those with low levels. Further, patients with high levels of cyclin A1 had significantly lower disease‐free survival (DFS) (P = 0.028). Multivariate analysis indicated that cyclin A1 protein expression was an independent prognostic factor for predicting DFS (P = 0.035) and OS (P = 0.045). No correlation between cyclin A1 expression and age was found. However, expression of cyclin A2, cyclin B1, cyclin E, CDK1, CDK2, p21 and p27 did not show prognostic significance in these AML patients.