Fine needle and core needle ultrasound guided biopsies for assessing suspected melanoma metastasis in lymph nodes and subcutaneous tissue.

BACKGROUND AND OBJECTIVE: The aim of this study was to investigate the diagnostic accuracy of ultrasound guided fine needle aspiration cytology (FNAC) and core needle biopsy (CNB) in different clinical scenarios for melanoma patients with lesions suspected of metastasis. METHODS: We included all patients at our department attending follow-up after surgery for cutaneous melanoma, who had undergone either FNAC or CNB between December 2016 and June 2019. Biopsy results were classified into one of four categories and verified with follow-up including imaging, re-biopsy or histology upon excision. The diagnostic accuracy of FNAC and CNB were calculated overall, and based on location of suspected metastasis, reason for suspicion and stage. RESULTS: We identified 232 biopsies in 164 patients; 109 FNACs and 123 CNBs. For FNAC, overall sensitivity was 83.3% and negative predictive value was 88.4%. For CNB, overall sensitivity was 92.4% and negative predictive value was 88.0%. There were significantly fewer nondiagnostic results using CNB compared to FNAC (χ1 2 = 6.7, p = 0.0095). CONCLUSIONS: There were no significant differences between the diagnostic accuracy of FNAC and CNB in the different clinical scenarios. We found significantly fewer nondiagnostic biopsies when using CNB, although this may reflect the type of lesions selected for each approach.

Estrogen receptor α is the major driving factor for growth in tamoxifen-resistant breast cancer and supported by HER/ERK signaling.

Resistance to tamoxifen is a major clinical challenge in the treatment of breast cancer; however, it is still unclear which signaling pathways are the major drivers of tamoxifen-resistant growth. To characterize resistance mechanisms, we have generated different tamoxifen-resistant breast cancer cell lines from MCF-7. In this model, we investigated whether signaling from human epidermal growth factor receptors (HERs), their downstream kinases, or from the estrogen receptor α (ERα) was driving tamoxifen-resistant cell growth. Increased expression of EGFR and increased phosphorylation of HER3 were observed upon acquisition of tamoxifen resistance, and the extracellular activated kinase (ERK) signaling pathway was highly activated in the resistant cells. The EGFR inhibitor gefitinib and the ERK pathway inhibitor U0126 resulted in partial and preferential growth inhibition of tamoxifen-resistant cells. All the tamoxifen-resistant cell lines retained ERα expression but at a lower level compared to that in MCF-7. Importantly, we showed via ERα knockdown that the tamoxifen-resistant cells were dependent on functional ERα for growth and we observed a clear growth stimulation of resistant cell lines with clinically relevant concentrations of tamoxifen and 4-OH-tamoxifen, indicating that tamoxifen-resistant cells utilize agonistic ERα stimulation by tamoxifen for growth. The tamoxifen-resistant cells displayed high phosphorylation of ERα at Ser118 in the presence of tamoxifen; however, treatment with U0126 neither affected the level of Ser118 phosphorylation nor expression of the ERα target Bcl-2, suggesting that ERK contributes to cell growth independently of ERα in our cell model. In support of this, combined treatment against ERα and ERK signaling in resistant cells was superior to single-agent treatment and as effective as fulvestrant treatment of MCF-7 cells. Together, these findings demonstrate that ERα is a major driver of growth in tamoxifen-resistant cells supported by HER/ERK growth signaling, implying that combined targeting of these pathways may have a clinical potential for overcoming tamoxifen resistance.

Association of tissue inhibitor of metalloproteinases-1 and Ki67 in estrogen receptor positive breast cancer.

BACKGROUND: The role of tissue inhibitor of metalloproteinases-1 (TIMP-1) in estrogen receptor (ER) positive breast cancer remains to be fully elucidated. We evaluated TIMP-1 as a prognostic marker in patients treated with adjuvant tamoxifen and investigated TIMP-1s association with Ki67 and ER/progesterone receptor (PR)/human epidermal growth factor receptor 2 (HER2) profiles. MATERIAL AND METHODS: TIMP-1 expression was evaluated by immunohistochemistry (IHC) on formalin fixed paraffin embedded primary tumor tissue in two independent cohorts comprised of 236 and 192 patients, respectively. RESULTS: No differences in disease free survival (HR 0.98; 95% CI 0.63-1.53; p = 0.92) and overall survival (HR 0.94; 95% CI 0.63-1.43; p = 0.79) were observed according to TIMP-1 status. A significant negative association between TIMP-1 and Ki67 was identified (p = 0.015). TIMP-1 expression did not differ significantly according to ER/PR/HER2 profiles. When analyzed as separate variables PR and HER2 status tended to have a positive but non-significant association with TIMP-1 (PR: p = 0.08; OR 2.54; 95% CI 0.91-7.10, HER2: p = 0.08; OR 0.48; 95% CI 0.21-1.08) whereas ER status was not associated with TIMP-1 expression (p = 0.48; OR 0.68; 95% CI 0.23-1.99). CONCLUSION: TIMP-1 does not appear to be prognostic in breast cancer patients receiving adjuvant tamoxifen. We identified a negative association between TIMP-1 and Ki67. We did not confirm our previous in vitro findings of a negative association between TIMP-1 and PR.

Prognostic value of Bcl-2 in two independent populations of estrogen receptor positive breast cancer patients treated with adjuvant endocrine therapy.

INTRODUCTION: Estrogen receptor (ER) status is not an optimal marker for response to adjuvant endocrine therapy since approximately 30% of patients with ER-positive tumors eventually relapse. Bcl-2 is regulated by ER and may thus be considered as an indicator of ER activity and a candidate supplementary marker to ER status. PATIENTS AND METHODS: Tumor tissue from 257 patients with ER-positive breast cancer treated with tamoxifen was used for determination of the best threshold for immunohistochemical Bcl-2 assessment as prognostic marker. Subsequently, samples from the Danish patients of the randomized clinical trial BIG 1-98 comprising 1191 ER-positive patients treated with tamoxifen, letrozole or a sequence of the two were immunohistochemically stained for Bcl-2 to further explore the prognostic value of Bcl-2. RESULTS: Two Bcl-2 levels were found to divide the population of the primary study into significantly different groups according to disease-free survival (DFS). Multivariate analysis confirmed the significance of the lowest level, and showed Bcl-2 to be an independent prognostic marker. Analysis of the Danish cohort of the BIG 1-98 confirmed that Bcl-2 was a significant predictor of DFS, independent of known prognostic markers. However, in an additional analysis of a subset of the Danish cohort of BIG 1-98 including only HER-2 normal patients, the effect of Bcl-2 was not statistically significant. DISCUSSION: Low Bcl-2 can predict poor outcome of patients with ER-positive tumors treated with adjuvant endocrine therapy, whereas the use of Bcl-2 for determination of addition of chemotherapy was not supported by this study.

Quantifying mast cells in bladder pain syndrome by immunohistochemical analysis.

OBJECTIVES: To evaluate a simple method for counting mast cells, thought to have a role in the pathophysiology of bladder pain syndrome (BPS, formerly interstitial cystitis, a syndrome of pelvic pain perceived to be related to the urinary bladder and accompanied by other urinary symptoms, e.g. frequency and nocturia), as >28 mast cells/mm(2) is defined as mastocytosis and correlated with clinical outcome. PATIENTS AND METHODS: The current enzymatic staining method (naphtolesterase) on 10 microm sections for quantifying mast cells is complicated. In the present study, 61 patients had detrusor biopsies taken between 2002 and 2005; the patients were given a clinical score, and sections of the biopsy stained with (i) naphtolesterase on 10 microm sections, staining every third section, or (ii) immunohistochemically with antitryptase on both 10 microm and 3 microm sections, with two and six unstained sections between, respectively. Mast cells were counted according to a well-defined procedure. RESULTS: The old and the new methods, on 10 and 3 microm sections, showed a good correlation between mast cell counts. When using tryptase staining and 3 microm sections, the mast cell number correlated well with the clinical score (Spearman's rho 0.576; 95% confidence interval 0.155-0.820) and 27 mast cells/mm(2) was the threshold suggesting mastocytosis. CONCLUSIONS: We recommend taking biopsies from the detrusor of patients with suspected BPS and examining them with tryptase-stained 3 microm thick sections, with every seventh section used for quantification; 27 mast cells/mm(2) is considered indicative of mastocytosis.

Carboplatin treatment of antiestrogen-resistant breast cancer cells.

Antiestrogen resistance is a major clinical problem in current breast cancer treatment. Therefore, biomarkers and new treatment options for antiestrogen-resistant breast cancer are needed. In this study, we investigated whether antiestrogen­resistant breast cancer cell lines have increased sensitivity to carboplatin, as it was previously shown with cisplatin, and whether low Bcl-2 expression levels have a potential value as marker for increased carboplatin sensitivity. Breast cancer cells resistant to the pure antiestrogen fulvestrant, and two out of four cell lines resistant to the antiestrogen tamoxifen, were more sensitive to carboplatin treatment compared to the parental MCF-7 cell line. This indicates that carboplatin may be an advantageous treatment in antiestrogen­resistant breast cancer; however, a marker for increased sensitivity would be needed. Low Bcl-2 expression was correlated with increased carboplatin response in the tamoxifen­resistant cell line MCF-7/TAMR-1 and overexpression of Bcl-2 in this cell line resulted in significantly reduced carboplatin sensitivity, confirming the anti-apoptotic role of Bcl-2. However, neither Bcl-2 expression alone, nor Bcl-2 in combination with Bcl-xL and Bax, could explain the observed responses to carboplatin in all tamoxifen­resistant cell lines, indicating that more markers are needed to predict the response to carboplatin in tamoxifen­resistant breast cancer.

Activated HER-receptors in predicting outcome of ER-positive breast cancer patients treated with adjuvant endocrine therapy.

The four human epidermal growth factor receptors (HER1-4) are involved in growth stimulation and may play a role in endocrine resistance. The receptors form dimers, leading to activation by mutual phosphorylation. Our purpose was to explore the role of the activated receptors (pHER1, pHER2, pHER3) in endocrine treated breast cancer in terms of co-expression and association with disease-free survival (DFS) in 1062 patients with ER-positive tumors. Furthermore, HER2 amplification was evaluated. We found positive associations between the phosphorylated receptors. pHER1 and pHER3 were co-expressed with one or two of the other activated receptors in 85% and 89% of tumors, respectively, whereas pHER2 was co-expressed with the other activated receptors in 54% of tumors. Except for HER2, which was associated with poor prognosis, none of the remaining markers were associated with DFS. However, frequent co-expression indicates a role of the other HER-family members in activation of HER2.