Şefik Güran

Hereditary TP53 Codon 292 and Somatic P16INK4A Codon 94 Mutations in a Li-Fraumeni Syndrome Family

Li-Fraumeni syndrome is an autosomal dominant disorder that is characterized by various types of cancer in childhood and adult cases. Although hereditary TP53 mutation is very rare in different human cancers, it has been frequently reported in Li-Fraumeni syndrome. On the other hand, hereditary mutations of TP57KIP2, P15INK4B, and P16INK4A, which affect the cell cycle similar to TP53, were observed in some types of cancer. In a Turkish family with the diagnosis of Li-Fraumeni syndrome, we analyzed the mutation pattern of TP53, P57KIP2, P15INK4B, and P16INK4A in the peripheral blood, and loss of heterozygosity (homo/hemizygous deletion) pattern of TP53 and P15INK4B/P16INK4A in two tumor tissues. The propositus had a seminoma, his daughter a medulloblastoma, and one of his healthy cousins, a TP53 codon 292 missense point mutation (AAA-->ATA; Lys-->Ile) in the peripheral blood cells. Tumor tissue obtained from the propositus with the seminoma revealed loss of heterozygosity in the TP53 gene. In the analyses of tumor tissues from the propositus and his daughter, a P16INK4A codon 94 missense point mutation (GCG-->GAG; Ala-->Glu) was observed with the hereditary TP53 mutation. P16INK4A codon 94 mutation observed in our family is a novel mutation in Li-Fraumeni syndrome. No other gene alteration in TP53, P57KIP2, P15INK4B, and P16INK4A was observed. Existence of the P16INK4A mutation and the hereditary TP53 mutation with or without loss of heterozygosity in the TP53 gene (seminoma/medulloblastoma) may be evidence for a common mechanism involved in tumorogenesis. The gene alterations in TP53 and P16INK4A genes may be used as tumor markers in our family.

Germline mutations of BRCA1 and BRCA2 genes in Turkish breast, ovarian, and prostate cancer patients

Distribution and prevalence of germline mutations in BRCA1 and BRCA2 differ among different populations. For the Turkish population, several studies have addressed high-risk breast cancer and ovarian cancer (BC-OC) patients. In most studies, both genes were analyzed in part, and a quite heterogeneous mutation spectrum was observed. For high-risk Turkish prostate cancer (PCa) patients, however, there are no data available about mutations of germline BRCA genes. To accurately determine the contribution of germline mutations in BRCA1 and BRCA2 in Turkish BC, OC, and PCa high-risk patients, 106 high-risk BC-OC patients, 50 high-risk PCa patients, and 50 control subjects were recruited. The study represents the only full screening, to date, of a large series of Turkish high-risk BC-OC patients and the only study in Turkish high-risk PCa patients. Mutation screenings were performed on coding exons of both genes with either denaturing gradient gel electrophoresis or denaturing high performance liquid chromatography, or with both techniques. Three deleterious mutations in BRCA1 and three deleterious mutations in BRCA2 were detected in different BC-OC patients, and one truncating mutation was detected in a high-risk PCa patient. In addition, 28 different unclassified and mostly novel variants were detected in both genes, as well as several silent polymorphisms. These findings reflect the genetic heterogeneity of the Turkish population and are relevant to genetic counseling and clinical management.

Genomic large rearrangement screening of BRCA1 and BRCA2 genes in high-risk Turkish breast/ovarian cancer patients by using multiplex ligation-dependent probe amplification assay.

In this study, MLPA assay was performed for detection of large rearrangements of BRCA1 and BRCA2 genes in 16 familial, 29 early onset, 3 male breast cancer, and 2 bilateral breast/ovarian cancer high risk Turkish index cases. MLPA assay for all exons of both genes and for 1100delC variant of CHEK2 gene were performed. Analyses, revealed no large genomic rearrangements in both genes, and, no 1100del variant in CHEK2 gene. Our data which represents the first results for Turkish patients, suggest that, the frequency of BRCA1 and BRCA2 genes’ large rearrangements is very low.

Familial Multiple Myeloma Associated with Disorders of Chronic Inflammation: First Report from Turkey

Multiple myeloma (MM) is a malignancy arising from mature plasma cells in the bone marrow and usually presents with bone destruction, hypercalcemia, anemia, renal damage, and increased susceptibility to infection. The etiology of MM is unknown, with no established lifestyle, occupational, or environmental risk factors. Because MM is an uncommon disease, etiologic assessments can be difficult. It has been reported to be in association with sarcoidosis, and in a few cases, rheumatoid arthritis. Familial type of MM with an autosomal dominant heredity pattern has also been reported. The genetic loci affected in these cases are still unknown. Herein we present a family with 3 affected cases in an autosomal dominant inheritance pattern. The first case was a man diagnosed to have immunoglobulin (Ig)A-type myeloma at the age of 50. The history revealed 2 more cases in the family: an uncle diagnosed to have unsecretory-type myeloma at the age of 76 and a cousin (the daughter of the affected uncle) who was diagnosed at the age of 48 years to have IgG-type myeloma and did not respond to therapy. This patient also had a history of sarcoidosis preceding the diagnosis of myeloma. All other affected family members had been treated for dental-oral infection (including chronic gingivitis) for 3 and 4 years before the diagnosis of myeloma. Karyotype analysis revealed pseudohypodiploidy and deletion of chromosome 13q in only the patient with coexisting sarcoidosis. To our knowledge, this is the first report on familial myeloma from Turkey. This family enhances the role of hereditary factors and chronic inflammation in the etiology of MM.

UNILATERAL TUBAL ECTOPIC TWIN PREGNANCY

Ectopic pregnancies develop when conception products implant somewhere other than in the endometrial cavity. If these pregnancies are not diagnosed and treated early, they can affect fertility and threaten the lives of otherwise healthy women. Twin pregnancies develop in approximately 1 in every 80 spontaneous pregnancies, and heterotopic pregnancies occur in about 1 in every 7,000 spontaneous pregnancies [1]. The probability of developing a unilateral tubal twin pregnancy has been calculated as 1 in every 125,000 spontaneous pregnancies. Most unilateral tubal twin gestations are reported to be monochorionic and monoamniotic [2]. A 24-year-old patient with a 5-week history of amenorrhea was admitted to our clinic with a complaint of left pelvic pain and vaginal bleeding which had lasted for 3 days. At admission, the patient was generally well and was conscious, cooperative and well-oriented. Her medical history revealed that she had been married for 3 months and was nulligravid, but used no contraception and had been planning on having children. The patient’s blood pressure was 120/80 mmHg, her pulse was 84 beats/minute, and her temperature was 37°C. Pelvic examination revealed extrauterine vaginal bleeding, closed cervical os, pain on cervical movement during manual examination, and left adnexal fullness and sensitivity. Abdominal examination revealed only left lower quadrant sensitivity. The left ovary appeared normal, but transvaginal ultrasound demonstrated an approximately 4 cm-sized left juxtauterine mass and free fluid in the left paraovarian area. Laboratory findings included a β-human chorionic

Methylation of SOCS3 in Myeloproliferative Neoplasms and Secondary Erythrocytosis/Thrombocythemia

Objective: Myeloproliferative neoplasms (MPNs) like essential thrombocythemia (ET), polycythemia vera (PV), and primary myelofibrosis (PMF) are acquired clonal hematopoietic stem cell disorders and originate from a multipotent hematopoietic stem cell. The SOCS1 and SOCS3 genes are negative regulators of the JAK/STAT signal pathway. In this study we investigate the promoter methylation of these genes in the pathogenesis of MPNs and secondary erythrocytosis/thrombocythemia. Materials and Methods: Promoter methylation of SOCS1 and SOCS3 genes was analyzed with methylation-specific PCR. PCR products were analyzed by agarose gel electrophoresis. Results: No disease-specific CpG island methylation of SOCS1 was observed. Hypermethylation of the SOCS3 promoter was identified in 5 out of 19 (26.3%) PV cases, 2 out of 21 (9.5%) ET cases, 1 out of 5 (20%) PMF cases, and 9 out of 42 (21.4%) cases of secondary erythrocytosis/thrombocythemia. Conclusion: The results revealed that promoter methylation of the SOCS3 gene suggests a possible role for SOCS3 methylation in the pathogenesis of MPNs and secondary erythrocytosis/thrombocythemia. Conflict of interest:None declared.

Usnik asitin tümör önleyici rolü mitokondrial apopitotik yolak üzerindedir

Health Sciences University, Gülhane Medical Faculty, Department of Medical Biology, Ankara, Turkey Gazi University, Faculty of Science, Department of Biology, Ankara, Turkey Corresponding author: Şefik Güran, Department of Medical Biology, Gülhane Medical Faculty, Ankara, Turkey E-mail: sefguran@yahoo.com Received/Accepted: April 10, 2017 / June 01, 2017 Conflict of interest: There is not a conflict of interest.

Usnic acid uses mitochondiral apopitotic pathway in its antitumoral role

SUMMARY Usnic acid is a secondary metabolite in lichens whose role has not been completely elucidated. Lichen extracts containing usnic acid have been utilized in medicine, perfumery, cosmetics, and ecology. In a recent manuscript, usnic acid was reported as an inhibitor in cell growth, inducing the cell cycle arrest and apoptosis in human lung carcinoma A549 cells. Herein we analyzed the antitumoral effect of usnic acid on the same cell line. Similar results were observed as in literature. We analyzed the gene expression results in the same panel which have possible role on cell apopitosis. As a result, usnic acid has stimulatory effect on APOPT1, CYCS, APAF1, CASP3, CASP9 gene expressions in our experiments. So, usnic acid has antitumoral effect on cancer cells, affecting on mithocondrial apopitotic genes. Key words: Usnic acid, cell growth, apopitosis, gene, antitumoral affect. OZET Usnik asit likenlerde bulunan halen rolu tam olarak ortaya konamamis bir sekonder metabolittir. Usnik asit iceren liken ekstraktlari tipta, parfum yapiminda, kozmetikte ve ekolojide kullanilmaktadir. Son bir yazida, insan akciger kanser A549 hucre hattinda usnik asit hucre dongusunun durdurulmasi ve apopitozis yoluyla hucre buyumesini durduran bir etken olarak tanimlanmistir. Calismamizda ayni hucre hattinda usnik asitin antitumoral etkisi bakilmistir. Literaturde olan ile benzer sonuclar ortaya konmustur. Ayni panel uzerinde hucre apopitozunda rolu olan genlerin ekspresyon analizine bakildi. Sonuc olarak, usnik asitin APOPT1, CYCS, APAF1, CASP3, CASP9 genleri uzerinde uyarici etkisi oldugu saptandi. Veriler usnik asitin kanser hucreleri uzerinde mitokondrial apopitotik genler araciligi ile antitumoral etki yaptigini ortaya koymaktadir. Anahtar Kelimeler: Usnik asit, hucre buyumesi, apopitoz, gene, antitumoral etki. Correspondance To: Prof. Dr. Şefik Guran Saglik Bilimleri Universitesi, Gulhane Tip Fakultesi, Tibbi Biyoloji AD. 06018 Etlik/Kecioren-ANKARA Phone: 03123043551

KANTARON YAĞI VEGFA, VEGFB, VEGFC ve FGF2 GENLERİ ÜZERİNDEN ERKEN DÖNEM KULLANILDIĞINDA YARA İYİLEŞMESİNDE OLUMLU ETKİ GÖSTERMEKTEDİR

Kantaron yagi Turkiye’ de yillardir geleneksel olarak kullanilmaktadir. Kantaron yaginin yara iyilestirici, agri kesici, enfeksiyon engelleyici ozellikleri bulunmaktadir. Literaturde kantaron yaginin ozellikle yara iyilesmesine olan pozitif etkisi ile ilgili bircok calisma vardir. Ancak simdiye dek bu ozelliklerin hangi yolaklar uzerinden oldugunu gosterir hayvan deneyleri iceren detayli bir calisma bulunmamaktadir. Calismamizda kantaron yaginin yara iyilesmesi uzerine erken donem (ilk 7 gun) ve gec donem (ikinci 7 gun) etkisi fareler (mus musculus) uzerinde steril kesiler olusturulup sutur edilerek gosterilmistir. Farelerde olusturulan yara yuzeyinde morfolojik olarak kantaron yaginin erken donem kullanimda pozitif etkisi olmakta, gec donem uygulamada bu etki gorulmemektedir. Bu bulgu kantaron yaginin yara iyilesmesine yara olustuktan sonraki ilk haftada pozitif etkisi oldugu seklinde yorumlanmistir. Bu donemde alinan dokulardan RT-PCR yontemi ile anjiyogenez uzerinden yara iyilesmesinde rolu olan VEGFA, VEGFB, VEGFC, PDGFB ve FGF2 genlerinin gen ekspresyon profilleri incelenmistir. Sonuc olarak erken donemde VEGFA, VEGFB, VEGFC ve FGF2 genlerinin gen ekspresyonlari artmakta, gec donemde bu etki yalnizca VEGFA geninde saptanmaktadir. Erken ve gec donemde bulunan gen ekspresyonu bulgulari saptanan morfolojik ozelliklerle korelasyon gostermektedir. Bulgular kantaron yaginin erken donem yara iyilesmesini anjiyogenezde rol oynayan VEGFA, VEGFB, VEGFC ve FGF2 uzerinden yaptigini gostermesi acisindan onemlidir.

Low Zip 4 gene expression levels in RPMI - 8226 and ARH - 77 cell lines support the possible role of zip 4 transporter protein in plasma cell tumorogenesis

Abstract Aim. Multiple myeloma and plasma cell leukemia are cancers of plasma cells. Multiple myeloma rarely transforms to plasma cell leukemia during the progression period. Zinc as a chemical element modulates proliferation and differentiation of cells by affecting several growth factors. Zip 4 modifies zinc metabolism in a cell as a transporter protein. While high Zip 4 gene expression was found in pancreas and hepatocellular carcinoma, low Zip 4 gene expression was observed in prostate carcinoma. Methods. Here, Zip 4 expression levels were studied in RPMI - 8226 and ARH - 77 cell lines as examples of multiple myeloma and plasma cell leukemia, respectively. Results. We found lower Zip 4 gene expression levels in both cell lines than that of the normal control (0,000157 in RPMI - 8226, 0,000227 in ARH - 77 cell lines and 1 in normal control) The findings were statistically significant (P < 0.05). The expression levels of Z ip 4 gene in both cell lines were approximately similar. No statistical significance was found between the expression levels of Zip 4 in both cell lines (P = 0.547). Conclusion. The results of this study support the possible role of Zip 4 gene expression in plasma cell dyscrasias. The similar result of Zip 4 gene expression level in both cell lines has no role in the transformation of multiple myeloma to plasma cell leukemia. Keywords: Zip 4 gene, gene expression, multiple myeloma, plasma cell leukemia Ozet Amac. Mltipl myelom ve plazma hucreli losemi, plazma hucresinden koken alan kanserlerdir. Hastaligin ileri evresinde multipl myelom nadiren plazma hucreli losemi formuna doner. Kimyasal bir element olarak cinko farkli buyume faktorleri uzerinden memeli hucrelerinde cogalma ve farklilasmayi duzenler. Tasiyici bir protein olarak Zip 4 ise hucrede cinko metabolizmasinda aktif rol alir. Pankreas ve karaciger kanserlerinde artmis Zip 4 gen ekspresyonu bulunurken, prostat kanserinde dusuk Zip 4 gen ekspresyonu tanimlanmaktadir. Yontemler. Bu calismada, sirasi ile multipl myelom ve plazma hucreli losemilere ornek olabilecek RPMI - 8226 ve ARH - 77 hucre hatlarinda Zip 4 gen ekspresyon duzeyleri bakilmistir. Bulgular. Her iki hucre hattinda Zip 4 gen ekspresyon duzeyi normale gore daha dusuktur (RPMI - 8226 hucre hattinda 0,000157 ve ARH - 77 hucre hattinda 0,000227). Elde edilen veriler istatistiksel olarak anlamlidir (P < 0,05). Her iki hucre hattinda Zip 4 gen ekspresyon duzeyleri birbirine cok yakin olarak bulunmustur. Aralarinda istatistiksel olarak anlamli bir fark da bulunmamistir (P = 0,547). Sonuc. Sonuclar plazma hucre diskrazilerinde Zip 4 gen ekspresyonunun olasi rolunu destekler niteliktedir. Her iki hucre hattinda birbirine yakin bulunan Zip 4 gen ekpresyon duzeyinin multipl myelomanin plazma hucreli losemiye donusmesinde rolu olmadigini ortaya koymaktadir. Anahtar sozcukler: Zip 4 geni, gen ekspresyonu, multipl myelom, plazma hucreli losemi