Handan Kaya

Voltage-Gated Sodium Channel Expression and Potentiation of Human Breast Cancer Metastasis

Purpose: Ion channel activity is involved in several basic cellular behaviors that are integral to metastasis (e.g., proliferation, motility, secretion, and invasion), although their contribution to cancer progression has largely been ignored. The purpose of this study was to investigate voltage-gated Na+ channel (VGSC) expression and its possible role in human breast cancer. Experimental Design: Functional VGSC expression was investigated in human breast cancer cell lines by patch clamp recording. The contribution of VGSC activity to directional motility, endocytosis, and invasion was evaluated by in vitro assays. Subsequent identification of the VGSC α-subunit(s) expressed in vitro was achieved using reverse transcription-PCR, immunocytochemistry, and Western blot techniques and used to investigate VGSCα expression and its association with metastasis in vivo. Results: VGSC expression was significantly up-regulated in metastatic human breast cancer cells and tissues, and VGSC activity potentiated cellular directional motility, endocytosis, and invasion. Reverse transcription-PCR revealed that Nav1.5, in its newly identified “neonatal” splice form, was specifically associated with strong metastatic potential in vitro and breast cancer progression in vivo. An antibody specific for this form confirmed up-regulation of neonatal Nav1.5 protein in breast cancer cells and tissues. Furthermore, a strong correlation was found between neonatal Nav1.5 expression and clinically assessed lymph node metastasis. Conclusions: Up-regulation of neonatal Nav1.5 occurs as an integral part of the metastatic process in human breast cancer and could serve both as a novel marker of the metastatic phenotype and a therapeutic target.

Distribution pattern of the Ki67 labelling index in breast cancer and its implications for choosing cut-off values

The Ki67 labelling index (LI - proportion of staining cells) is widely used to reflect proliferation in breast carcinomas. Several cut-off values have been suggested to distinguish between tumours with low and high proliferative activity. The aim of the current study was to evaluate the distribution of Ki67 LIs in breast carcinomas diagnosed at different institutions by different pathologists using the method reflecting their daily practice. Pathologists using Ki67 were asked to provide data (including the LI, type of the specimen, receptor status, grade) on 100 consecutively stained cases, as well as details of their evaluation. A full dataset of 1709 carcinomas was collected from 19 departments. The median Ki67 LI was 17% for all tumours and 14% for oestrogen receptor-positive and HER2-negative carcinomas. Tumours with higher mitotic counts were associated with higher Ki67 LIs. Ki67 LIs tended to cluster around values ending with 5 or 0 both in cases where the values were obtained by counting the proportion of stained tumour cell nuclei and those where the values were obtained by estimation. On the basis of the distribution pattern described, some currently used Ki67 LI cut off values are not realistic, and it is proposed to select more realistic values ending with 0 or 5.

Could prolactin receptor gene polymorphism play a role in pathogenesis of breast carcinoma

SUMMARY Objective: Constitutive activation of various hormone and growth factor receptors is newly recognised as a common cause of tumour development. This study investigated the presence of any mutation or polymorphism of prolactin receptor (PRLR) in 38 patients with breast cancer. Research methods: Genomic DNA was extracted and PCR amplification was carried out for exon 1–10 of PRLR from tumoral and adjacent non-cancerous breast tissue of tumour specimens from 38 breast cancer patients. PCR products were analysed by SSCP and automatic sequencing for mutations. Results: For the first time, A150C (Leu→Ile) transversion at exon 6 of PRLR in tumour tissues, in adjacent non-cancerous breast tissues, and in blood samples of two (5.3%) out of 38 patients with breast cancer were detected. In contrast to this finding, no polymorphism of PRLR in blood samples of 100 normal individuals were found. Conclusion: Polymorphism of prolactin receptors might play a role in mammary carcinogenesis as a consequence of intracellular changes of PRLR signalling.

Mitochondrial DNA common deletion is not associated with thyroid, breast and colorectal tumors in Turkish patients

Recently, efforts have been focused on mitochondrial DNA changes and their relation to human cancers. Among them, a 4977 bp deletion of mitochondrial DNA, named “common deletion”, has been investigated in several types of tumors, with inconsistent results. In this study, we investigated the presence of the common deletion in tissues from 25 breast, 25 colorectal and 50 thyroid tumors and in the adjacent healthy tissues from Turkish patients. Samples from healthy volunteers were also evaluated for comparison. Two PCR-based methods were used for the detection of the common deletion. First, two pairs of primers were used to amplify wild-type and deleted mtDNA. Then, a highly sensitive nested-PCR was performed, to determine low amounts of deleted genomes. By the first method, wild-type mtDNAs were observed in all samples, but a deletion was observed in only six thyroid samples, by using the nested-PCR method. In conclusion, the mitochondrial common deletion was very rare in our study group and did not appear to be not related with cancer.

A Giant Adrenal Pseudocyst: Case Report and Review of the Literature

A 45-year-old woman whose MR images revealed a lobulated, complicated cyst with septations on the superior pole of left kidney underwent retroperitoneoscopic cyst marsupialization. In pathologic examination beneath the atrophic adrenal gland, hyalinized, calcification foci cyst wall without any surrounding epithelium was seen microscopically. It was reported to be “adrenal pseudocyst”.

Her-2/neu gene amplification compared with HER-2/neu protein overexpression on ultrasound guided core-needle biopsy specimens of breast carcinoma.

Genomic amplification and oncoprotein overexpression of Her-2/neu was studied on ultrasound core needle biopsy specimens of the infiltrative ductal carcinomas of the breast. We performed “two colour” fluorescence in situ hybridization (FISH) for Her-2/neu and chromosome 17 and compared the FISH results with the immunohistochemical overexpression of Her-2/neu protein by 2 antibodies (DAKO HercepTest and the BioGenex monoclonal antibody AM 134-5M). Furthermore, following radical mastectomy with axillary dissection, Her-2/neu status of the patients were compared with the well known histopathological prognostic factors such as histologic grade, tumor stage, lympho/ vascular invasion, surgical margin status and Paget’s disease. Amplification was demonstrated 27% of the cases. Her-2/neu protein overexpression was detected in 47% and 80% of the cases with CB11 and HercepTest respectively. We revealed statistically significant association between the tumor, oncoprotein expression and oncogene amplification (p<0.05). The results of our study showed that combination of IHC and FISH methods enhances the evaluation of tumor genetics at both gene and protein level for the analysis of Her-2/neu in breast carcinoma.

Solitary fibrous tumor of the kidney

Solitary fibrous tumors are rare spindle cell neoplasms usually arising in the pleura. They have, however, also been reported at extrapleural locations. Solitary fibrous tumor (SFT) of the kidney is rare. Despite its rarity, histological diagnosis of solitary fibrous tumor is crucial to avoid misdiagnosis with other more aggressive tumors arising in the kidney. We report a solitary fibrous tumor of the left kidney that presented as a malignant tumor in a 51‐year‐old woman, and include clinical and radiographic findings. The tumor was well circumscribed and composed of spindle cells in a collagenous stroma. Immunohistochemistry showed reactivity for vimentin, CD 34, BCL‐2 protein and CD99. Immunohistochemical stains for cytokeratin, S‐100, desmin, α‐smooth muscle actin and HMB‐45 were negative. A diagnosis of SFT was made based on light microscopy and immunohistochemistry.

Apocrine Differentiation in Invasive Pleomorphic Lobular Carcinoma with In Situ Ductal and Lobular Apocrine Carcinoma: Case Report

Invasive pleomorphic lobular carcinoma (PLC) is a distinctive aggressive subtype of invasive lobular carcinomas (ILC). We report one case of PLC with in situ PLC and ductal carcinoma in situ with apocrine features.

Collagen Ultrastructure and TGF-β1 Expression Preserved with Aminoguanidine During Wound Healing in Diabetic Rats

Advanced glycoxidation end products have been implicated in delayed diabetic wound healing. In this study, we evaluated the effects of aminoguanidine, which is an advanced glycation and nitric oxide (NO) synthase inhibitor, on extracellular matrix protein expression, collagen configuration, and nitrite/nitrate levels in wounds of diabetic rats. Sixteen Wistar male rats were made diabetic by streptozotocin. Of these, eight rats were given AG (aminoguanidine bicarbonate (AG) (group DAG) in their drinking water, and eight rats were followed as diabetic paired controls (group D). Eight healthy rats were followed as the healthy control group (group H). At the eighth week, a 2 × 2 cm area full-thickness skin defect was created. The degree of contraction of the open wounds was evaluated for 2 weeks duration. On the 15th postoperative day, wound surface areas were measured, and wound specimens and blood samples were collected. The shrinking percentage of the wounds was small in both groups H and DAG compared with group D (p < 0.05). Similar to healthy rats, the aminoguanidine-treated diabetic rats had very strong transforming growth factor (TGF)-β1 expression in granulation tissue and intact skin in comparison with diabetic controls. In the diabetic group, the intact skin demonstrated sparsely distributed regular collagen fibers in the granulation zone, and the regular pattern of collagen fibers was lost. In conclusion, aminoguanidine improves wound healing, restores growth factor TGF-β1 expression, and preserves collagen ultra structure, whereas it has no prominent effect on NO levels within wound tissue in diabetic rats.

Incidental carcinoid of appendix in cesarean section

A case with carcinoid tumor of the appendix was encountered incidentally during an elective cesarean section. The tumor was discovered as a result of the routine exploration of the whole abdomen. The investigation for metastasis proved no evidence of spread and the patient was treated solely by surgery.