Aydan Eroğlu

Risk factors related to locoregional recurrence in squamous cell carcinoma of the skin

A retrospective analysis was performed to identify the risk factors associated with development of locoregional recurrent disease in patients with primary squamous cell carcinoma of the skin. Step‐wise logistic regression analysis was used in this study, which consisted of 1,039 patients treated from January 1980 to December 1989 at Ankara Oncology Hospital. Locoregional recurrence occurred in 187 (18%) of these patients within a mean disease‐free period of 15 months. Age, sex, anatomical location, size of lesion, lymph node status at diagnosis, stage according to TNM classification, histopathologic grade, previous therapy, treatment modality, lesions arising from scar tissue (scar carcinoma), regional lymph node dissection, concomitant premalignant tumor of the skin, development secondary nonmelanotic skin carcinoma, and second malignancy were used as variables that could be correlated with locoregional recurrent disease. No correlation was found between development of recurrence and previous therapy, second nonmelanoma skin cancer, second malignancy, premalignant skin tumor, sex, or regional lymph node dissection. Although univariate analysis demonstrated that location, tumor size, patients age, lymph node status, stage, histopathologic grade, scar carcinoma, and treatment modality were associated with an increased risk of locoregional recurrence, it was found that stage of the disease (P < 0.001), treatment modality (P < 0.01), tumor arising from scar tissue (P < 0.01), and histopathologic grade (P < 0.05) were statistically significant as risk factors of recurrence when a multivariate analysis was applied.

Dihydrofolate reductase (DHRF) 19-bp intron-1 deletion and methylenetetrahydrofolate reductase (MTHFR) C677T polymorphisms in breast cancer

The causes of breast cancer (BC) are not clearly known, although different genetic and environmental factors play role in its development. Methylenetetrahydrofolate reductase (MTHFR) is a key enzyme in folate-mediated onecarbon metabolism, regulating the DNA methylation and synthesis. Reduced MTHFR activity is associated with genomic DNA hypomethylation. MTHFR is suspected to play a role in the etiology of cancer, via its effects on DNA methylation and nucleotide synthesis [1]. The C to T substitution at nucleotide 677 of MTHFR gene converts an alanine to a valine is associated with reduced enzyme activity and causes increased plasma homocysteine levels. Associations of MTHFR C677T polymorphism with BC have been investigated [1–4]. Dihydrofolate reductase (DHFR) is also important folate metabolizing enzyme that is essential for DNA synthesis and homocysteine remethylation. Recently, Johnson et al. described a 19-bp deletion polymorphism in intron-1 of DHFR that might affect gene expression [5]. More recently, Gellekink et al. reported that 19-bp del/del genotype was associated with a lower plasma homocysteine level and suggested that 19-bp deletion might increase DHFR expression thereby facilitating homocysteine remethylation [6]. The aim of the present study was to evaluate the association of BC risk with MTHFR C677T and DHFR 19-bp deletion polymorphisms. The study consisted of 110 women with BC. Ninety-five healthy women (in matched control) were used as control. Genomic DNA isolation was performed from peripheral venous blood by standard phenol–chloroform extraction, and polymerase chain reaction of 19-bp deletion polymorphism in intron-1 of DHFR was performed according to previously described method [5], using the following primers: F 50-CCACGGTCGGGGTACCTGGG-30, F 50ACGGTCGGGGTGGCCGACTC-30 and R 50-AAAAGGG GAATCCAGTCGG-30. Amplification was made as follows: initial denaturation of one min at 94 C; followed by 35 cycles of 94 C/55 s, 64 C/55 s, 72 C/55 s, and final extension of 12 min at 72 C (Biometra, Germany). The resulting PCR products were 96-bp and 113-bp. Amplified products were subjected to 4% agarose gel. Genotype data were obtained from 179 subjects. MTHFR C677T polymorphism was determined by using commercially available LightCycler kit (Roche Diagnostic, Roche Molecular Biochemicals, Mannheim, Germany). Genotype data were obtained from 205 cases. The groups were compared using v test and P-value \ 0.05 was considered significant. No differences were observed in the distribution of MTHFR C677T genotype or allele frequencies in the BC cases versus control subjects (P [ 0.05; Table 1). A slightly increased frequency of TT genotype in cancer patients was found but this did not reach statistical significance. We did not find a significant association of DHFR 19-bp deletion polymorphism with BC risk. The frequency of 19-bp deletion allele was lower in BC patients than in control group. Various groups have evaluated the association between MTHFR C677T polymorphism and BC risk, though the findings were conflicting. Some authors observed an increased risk for the carriers of TT genotype. Recent R. Çam A. Eroglu (&) Department of General Surgery, Ankara University Medical School, Cebeci Kampus, Ankara 06590, Turkey e-mail: aydaneroglu@hotmail.com

The 19-bp deletion of dihydrofolate reductase (DHFR), methylenetetrahydrofolate reductase (MTHFR) C677T, Factor V Leiden, prothrombin G20210A polymorphisms in cancer patients with and without thrombosis

Venous thromboembolism (VTE) is a common complication in cancer patients. Several genetic risk factors related to thrombophilia are known; however, their contributions to thrombotic tendency in cancer patients have conflicting results. In the present study, we have focused on the prevalence of methylenetetrahydrofolate reductase (MTHFR) C677T, dihydrofolate reductase (DHFR) 19-bp deletion within intron 1, factor V Leiden (FVL), and prothrombin (PT) G20210A polymorphisms in cancer patients with and without VTE. The study consisted of 63 cancer patients with VTE (group 1) and 124 cancer patients who had no evidence of VTE (group 2). Four gene polymorphisms were determined by the method of polymerase-chain-reaction-based DNA analysis. The prevalence of DHFR 19-bp deletion and MTHFR C677T polymorphisms was similar in two groups (p > 0.05). The frequency of FVL was significantly higher in group1 compared with group 2 (31.7% vs. 1.6%, p < 0.0001), but PT G20210A polymorphism was not associated with VTE. Cancer patients with thrombosis should be evaluated for FVL, but routine screening for PT G20210A, MTHFR C677T and DHFR 19-bp deletion polymorphisms is not suggested.

Expression of c-kit proto-oncogene product in breast cancer tissues

BackgroundThe published results on expression of c-kit in benign and malignant breast tissues vary.Materials and MethodsThe immunohistochemical expression of c-kit proto-oncogene product in 52 invasive breast cancer tissues and 16 benign breast tumor (fibroadenoma) tissues was studied using anti-c-kit proto-oncogene product antibody. Its expression was evaluated by immunoreactive score (IRS).ResultsIn breast cancer tissues, the mean IRS of c-kit proto-oncogene product expression was significantly increased compared to those of fibroadenoma (3.4 ± 2 and 2.19 ± 1.8, respectively,p = 0.035). The mean IRS of c-kit expression was higher in the group comprising estrogen (ER) positive tumor than in the group of ER negative (4.1 ± 2.1 and 2.7 ± 1.8, respectively,p = 0.012) but no statistically significant relationship was seen between the expression of c-kit proto-oncogene product and other clinicopathological parameters of breast cancer, including histologic type, tumor size, lymph node metastasis, distant metastasis, stage, progesterone receptor, c-erbB-2 expression, menopausal status and age of the patient (p > 0.05).ConclusionsOur results show that a high level of c-kit expression occurs frequently in invasive breast cancer, and its expression is associated with ER but unrelated to other clinico-pathological variables.

Prevention of intra-abdominal adhesions by using Seprafilm® in rats undergoing bowel resection and radiation therapy

Intra‐abdominal adhesions are an important surgical problem. Colorectal operations are a major cause of adhesive obstruction. Radiation therapy (RT) is frequently used as an adjuvant therapy to surgery for rectal cancer, though its value for colon carcinoma remains unclear. Peritoneal injuries including the surgical trauma, tissue ischaemia as well as RT are associated with peritoneal fibrinolytic activity. A sodium hyaluronate and carboxymethylcellulose bioresorbable membrane (Seprafilm®) has been used to reduce intra‐abdominal adhesion formation. We have investigated the effect of Seprafilm® on intra‐abdominal adhesion in rats receiving RT after the resection of the left colon.

Factor V Leiden and PT G20210A mutations in cancer patients with and without venous thrombosis

*Department of General Surgery and Surgical Oncology, Numune State Hospital, Selc¸uklu, Konya, Turkey; and Departments of RadiationOncology, Pediatric Molecular Genetic and §General Surgery, Ankara University Medical School, Cebeci, Ankara, TurkeyTo cite this article: Eroglu A, Kurtman C, Ulu A, C¸ am R, Akar N. Factor V Leiden and PT G20210A mutations in cancer patients with,and withoutvenous thrombosis. J Thromb Haemost 2005; 3: 1323–4.

Retroperitoneal soft tissue sarcoma: effect of hyperthermic total abdominal perfusion.

Aims and Background Soft tissue sarcomas (STS) of the retroperitoneum are rare tumors. Surgery remains the principal modality of therapy in the management of primary and recurrent retroperitoneal STS. However, little is known about the effect of regional chemotherapy using hyperthermic total abdominal perfusion (HTAP). We analyzed independent prognostic variables in 33 patients with STS in the retroperitoneum admitted from November 1990 through December 1996. Methods and Study Design Data regarding patients’ age, gender, tumor size, histological tumor type, tumor location, type of operation (primary or secondary surgery), extent of surgical management (marginal or extended), use of HTAP, tumor grade, and tumor stage according to the TNM classification were examined by univariate and multivariate analyses. Results All 33 patients underwent complete resections (marginal or extended). Eleven of them received locoregional chemotherapy by HTAP. The overall cumulative 5-year survival rates in patients with stage IIA and advanced disease were 82% and 22%, respectively (log-rank test, P <0.01). Using Coxs proportional hazard model, tumor stage, use of HTAP and type of operation were found to have significant influence on overall survival (P <0.05). Conclusions Our results showed that complete resection along with HTAP chemotherapy may improve survival in patients with retroperitoneal STS. These phase II data could be used to support the initiation of a phase III trial to test HTAP in patients submitted to complete resection of retroperitoneal STS.

Factor V Leiden, prothrombin G20210A and methylenetetrahydrofolate reductase (MTHFR) C677T polymorphisms and the risk of tamoxifen-associated thromboembolism in breast cancer patients

Cancer produces a hypercoagulable state which may lead thrombosis. However, the effect of hereditary thrombophilia in cancer patients with thrombosis is still unclear. Some studies have been evaluated the role of factor V Leiden (FVL)mutation on the thrombosis risk in malignancy with conflicting results [1,2]. Cancer therapy can influence the risk of thrombosis aswell as type ofmalignancy, extent of disease. Cytotoxic chemotherapy increases the risk of vascular events in breast cancer patients, especially when combining chemotherapy with hormone therapy [3]. Tamoxifen has been widely used in the treatment of breast cancer. Women with breast cancer receiving tamoxifen have an increased risk of venous thromboembolism (VTE) by 2to 7-fold compared to placebo [4]. However the presence of FVL is not associated with thrombosis among women taking tamoxifen to prevent breast cancer [5]. Therefore FVL screening is not suggested to identify at increased risk of VTE for women with breast cancer risk taking tamoxifen. It was first time Weitz and coworkers reported two breast cancer patients with FVL who underwent VTE while taking tamoxifen [6]. After the publication, we evaluated the frequency of FVL and prothrombin (PT) G20210A polymorphisms in breast cancer patients treated with tamoxifen who developed VTE compared with those without VTE [7]. The nested case-control study was the first to show the association of the presence of FVL with VTE development among breast cancer patients taking tamoxifen, but not PT G20210A allele was found. Recently Garber et al have reported a large case-control study conducted from 34 centers to compare the frequency of FVL mutation in breast cancer patients with or without thrombosis while receiving adjuvant tamoxifen [8]. The authors demonstrated that FVL was significantly associated with VTE risk among breast cancer patients taking tamoxifen. Encouraged by the report, we have reevaluated the frequencies of FVL and PT G20210A polymorphisms in breast cancer patients receiving tamoxifen with and without VTE in the study including new added cases. In addition to these polymorphisms, we also investigated the effect of methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism on VTE risk in the present study. Sixteen of our breast cancer patients developed VTE while receiving adjuvant tamoxifen (group 1). Fifty women with breast cancer taking tamoxifen without VTE were included as group 2. Genomic DNA isolation was performed from peripheral venous blood by standard phenol-chloroform extraction and FVL, PT G20210A and MTHFRC677T polymorphisms were determined by using commercially available Light Cycler kits (Roche Diagnostic, Roche Molecular Biochemicals,

Factor V Leiden and prothrombin G20210A polymorphisms are not associated with disease-free survival in breast cancer.

It is known that cancer and its treatment can induce a hypercoagulable state. On the other hand, coagulation and fibrinolysis can play a significant role in tumor growth, invasion, dissemination and metastasis [1].The significant presence of factor V Leiden (FVL) and/or prothrombin (PT) G20210A polymorphisms which are the two most common defects in Caucasians are responsible for coagulation abnormality in thrombophilia. A few previous reports investigated the relationship between the cancer development and FVL and PT G20210A polymorphisms and found no association of FVL and some types of cancer [2–6]. On the contrary, Pihusch and coworkers demonstrated that the prevalence of PT G20210A polymorphism was significantly increased in patients with gastrointestinal carcinoma as compared to normal subjects [7]. However the role of FVL and PT G20210A polymorphisms in the recurrence risk and prognosis of breast cancer has not been investigated yet. We have read with great interest in the recent publications about experimental metastasis model in FVLdeficient mice [8, 9]. Bruggeman et al. showed that mice carrying FVL mutation were more susceptible to cancer cell metastasis after the injection of melanoma cells into the tail vein [8]. But, other study by Klerk et al. indicated that FVL mutation had no effect on the metastasis of colon cancer to the livers of mice [9]. Encouraged by the reports, we have retrieved data in 123 operable breast cancer and performed an analysis to investigate the role of FVL and PT G20210A polymorphisms in the recurrence (loco-regional and/or distant recurrence) of breast cancer. Genomic DNA isolation was performed from peripheral venous blood by standard phenol-chlorophorm extraction and FVL and PT G20210A polymorphisms were determined by using commercially available LightCycler kits (Roche Diagnostic, Roche Molecular Biochemicals, Mannheim, Germany) [10]. Data regarding patient’s age, tumor type, tumor size, lymph node status, tumor stage, grade, estrogen and progesterone receptor, c-erb B2 expression, FVL and PT G20210A polymorphisms were examined by univariate and multivariate analyses. The Kaplan Meier method was applied to examine the effect of individual variables on disease free survival (DFS). The significance of the observed difference between groups was calculated by the logrank test. The combined and independent effects of the factors on DFS were examined using the Cox proportional hazards model. Statistical analysis was done using SPSS 13.0 for Windows. For all test, a P value of less than 0.05 was considered to be significant. The follow-up period ranged from 4 to 216 months (mean, 57 months). Recurrent disease occurred in 22 patients within a mean time of 26 months (range: 6– 72 months). The univariate analysis factors having significant effects on DFS were tumor size, lymph node status, tumor stage, and c-erb B2 expression (P \ 0.05). FVL was detected in 10 breast cancer patients and the recurrent disease was developed in three of them. As shown in Fig. 1, the 5-year DFS rate in patients with FVL mutation was lower than those without FVL (68% vs. 82%), however this difference did not reach statistically significant A. Eroglu (&) R. Cam Department of General Surgery, Ankara University Medical School, Ankara, Turkey e-mail: aydaneroglu@hotmail.com

Deep venous thrombosis of the extremity diagnosed by color Doppler ultrasonography after isolated limb perfusion.

Aim Isolated limb perfusion (ILP) is used to treat locally advanced sarcoma and melanoma of the extremities. ILP is associated with a 1.7% to 10% incidence of deep venous thrombosis (DVT). The aim of the study was to examine the effect of ILP on the peripheral venous system and to evaluate the diagnostic accuracy of color Doppler ultrasonography (US). Methods A total of 26 patients with locally advanced sarcoma or melanoma of the extremities received a total of 38 ILP. The patients were evaluated preoperatively and postoperatively by color Doppler US as a noninvasive venous measurement. We used a color Doppler US system (SSA-270A, Toshiba) with a 7.5-MHz linear transducer for gray-scale imaging and a 5-MHz vascular transducer for color Doppler imaging. Results Seventeen patients underwent single ILP while the others were treated with multiple ILPs. Color Doppler US showed a reflux flow in three (7.9%) patients and DVT occurred subsequently in these patients. Conclusions Color Doppler US is a noninvasive and clinically useful diagnostic technique in the diagnosis of extremity DVT. We recommend anticoagulant therapy in patients with reflux on ultrasonographic examination.