Prunella vulgaris l. ve prunella grandiflora l.’den saflaştırılan rosmarinik asitin farklı tümör hücreleri üzerindeki sitotoksik aktivitesi

Rosmarinic acid was isolated from ethanol extractions obtained from Prunella grandiflora L. and P. vulgaris L. The total phenolic content of ethanol fractions during isolation was determined by the Folin-Ciocateu method. The cytotoxic doses of the isolated rosmarinic acid were determined by WST-1 (Roche Applied Sciences, Mannheim, Germany) cell proliferation assay. A 10-60ng cytotoxic dose was determined for pancreas (PANC-1), prostate (PC-3), colon (HT-29) and breast (MDAMB 436) cancers and GBM (T98G) cell lines and lymphatic tissues. 24 and 48h incubation periods were applied during dose determinations. An antiproliferative effect was observed at the end of 48h incubation period with 50ng rosmaniric acid treatment in PC3 cell line and with 60ng treatment in PANC-1, HT-29, MDA-MB 436 and T98G cell lines. No cytotoxic activity of rosmarinic acid was observed in non-tumor cells.Bu çalışmada rosmarinik asit bileşiği, Prunella grandiflora L. ve P. vulgaris L. türlerinden elde edilen etanol ekstraktlarından saflaştırılmıştır. Saflaştırma işlemi sırasında elde edilen metanol fraksiyonlarının toplam fenol içeriği FolinCiocalteu yöntemi ile belirlenmiştir. Prunella L. türlerinden saflaştırılan rosmarinik asitin farklı kanser hücreleri üzerinde WST-1 (Roche Applied Sciences, Mannheim, Almanya) yöntemiyle sitotoksik doz çalışmaları yapılmıştır. Buna göre pankreas (PANC-1), prostat (PC-3), kolon (HT-29) ve meme (MDA-MB 436) kanserleri ile GBM (T98G) hücre hatları ve lenf dokularında 10-60ng arasında sitotoksik doz belirlenmiştir. Sitotoksik doz belirleme çalışmaları için 24 ve 48 saat inkübasyon süreleri çalışılmıştır. 48 saat inkübasyon süresi sonunda PC3 hücre hattı için 50ng ve PANC-1, HT-29, MDA-MB 436 ve T98G hücre hatları için 60ng rosmarinik asit uygulamasında antiproliferatif etki gözlenmiştir. Sağlıklı hücrelerde rosmarinik asitin sitotoksik etkisi gözlenmemiştir

Cytogenetic investigations in shoe workers

Çalışmamızda biri kadın, diğerleri erkek olan toplam 58 ayakkabı işçisi ve rasgele olarak seçilmiş biç bir mutajen ve kanserojen ajana maruz kalmamış 20 kişilik kontrol grubu üzerinde periferik kan lenfositleriyle, sitogenetik analiz yöntemi kullanılmıştır. Her iki grupta da, gap, kırık, asentrik fragman, rearrangement ve poliploididen oluşan hücresel hasarın sıklığı araştırılmıştır. Kromozomal hasar (Özellikle kromatid gap ve kırıkları) kontrol grubuna oranla, etkilenen grupta anlamlı bir şekilde yüksek bulunmuştur. Ayakkabı işçileri ve kontrol grubunu oluşturan bireylerin sigara kullanımları ve alkol alışkanlıkları hem kendi içlerinde hem de diğer grupla karşılaştırılmış, her iki durumda da karşılaşılmış her iki kriterler arasında bir ilişki bulunamamıştır. Ayrıca, çalışmaları sırasında benzenden etkilenme süresi ile kromozomal hasarın sıklığı arasında da anlamlı bir ilişki bulunamamıştır.In the present study, the method of cytogenetic analysis of peripheral blood lymphocytes was used to investigate 58 (57 men and 1 female) shoe workers exposed to benzene, and 20 individuals selected from general population not exposed to particular mutagenic or carcinogenic agents (control group). Frequecies of damaged cells, including gap, break, acentric fragment and rearrangement were scored in both groups. The incidence of chromosomal aberrations (particularly chromatid gaps and breaks) in the exposed group were significantly increased when compared with the control group. Shoe workers and the individuals of control group were compared according to their smoking and dirinking habits both by themselves and eachother. As a result of those comparisons no correlation was found in the incidence of.chromosomal aberration in both situations. In the same way, there wasn't found any significantly relation between the working period in the group exposed to benzene frequency of chromosomal aberration.Bursa Ayakkabıcılar Derneğ

The Investigation of genetic predisposition in patients with colorectal cancer and their relatives

Frajil bölgeler kromozomlar üzerinde boya almayan özel gap ve kırık noktalandır ve onlar çeşitli kültür koşullan ile oluşturulabilirler. Çeşitli çalışmalar frajil bölge ekspresyonunu arttıran ve birçok klastojenik ajan olduğunu göstermiştir. Biz de çalışmamızda periferik kan lenfositlerinde gözlediğimiz prometafaz kromozomlarda common frajil bölgelerin ekspresyonunu afidikolin, bromodeoksiuridin ve kafein ile indükledik. Kromozomal aberasyonlar ve frajil bölge ekspresyonlan, 32 kolon kanserli hasta, 30 asemptomatik kolon akraba, 36 rektum kanserli hasta, 30 rektum akraba ve 30 da yaşlan hasta ve akraba grupları ile uyumlu, ailesinde kanser hikayesi bulunmayan sağlıklı kontrol bireylerinden oluşan toplam 158 vakada değerlendirildi. Hem hasarlı hücre oram (P0.05). Biz iki grubumuzda da apbidicolin tip common frajil bölgeler belirledik. Kolon kanserli hastalar ve yakınlarında belirlediğimiz frajil bölgeler lp36, lp31, lp21, lq21, lq25, lq44, 2p24, 2pl6, 2q21, 2q33, 2q37, 3p21, 3pl4, 15ql5, 5q21, 5q31, 13ql3 ve 14q24 şeklinde sıralanmaktadır. Bunlarm içinde lp21, lq21, lq25, lq44, 2p24, 2pl6, 2q33, 2q37, 3pl4, 5q21, 5q31 ve 14q24 frajil bölge oranları, kontrol grubu ile karşılaştırıldığında istatistiki olarak anlamlı bulundu (P0.05). We determined aphidicolin type common fragile sites in our both groups. These sites in patients with colon cancer and relatives were following : lp36, lp31, lp21, lq21, lq25, lq44, 2p24, 2pl6, 2q21, 2q33, 2q37, 3p21, 3pl4, 5ql5, 5q21, 5q31, 13ql3 and 14q24, In this fragile sites, lp21, lq21, lq25, lq44, 2p24, 2pl6, 2q33, 2q37, 3pl4, 5q21, 5q31 and 14q24sites were statistically significant when compared with control group (P<0.05-P<0.0001). Moreover expression of 2q33, 3pl4 and 5q21 sites were observed highest significant in all of fragile sites. Fragile sites in rectum cancered patients and their relatives were following: lp36,. Ip31, lp21, lq21, lq25, lq44, 2p24, 2q21, 2q33, 2q37, 3pl4, 5q21, 5q31, 13ql3, 14q24, 16q23 and 18q21. When compared with control group fragile sites of lp21, lq25, lq44, 2q33, 5q21, 5q31 and 14q24 were statistically significant (P<0.05-P<0.0005). Highest statistically significancy was observed in 2q33 in this group. Our results indicate that there is a relative increase in chromosomal aberrations and fragile sites expressions in patients with colorectal cancer and their relatives. These data also suggest that common fragile sites may really be preferential points of breakage and that then- expression might be primary Contibutors to chromosomal damages of the somatic genome. So this type studies may be helpful to early detection of cancer, clarifiy the biological mechanism of cancer development, progression and in the protection from cancer

An in vitro model for the development of acquired tamoxifen resistance

The development of resistance to tamoxifen (Tam) remains a challenging clinical problem for ER+ breast cancer patients. To understand the mechanisms underlying of resistance, previous studies have driven the acquisition of Tam resistance by exposing cells to varying concentration of drug for varying lengths of time. However, a detailed protocol for the establishment of Tam-resistant cells remains to be clarified. In the present study, we aimed to determine and compare the effect of different in vitro protocols on the degree of resistance to 4-hydroxytamoxifen (4-OH Tam) for MCF7 cells. For this purpose, MCF7-Tam resistance (MCF7-TamR) cells were developed by treated with different concentrations (100, 200, 400, 600, 800 and 1000 nM) of 4-OH Tam over 3 months. The relative resistance was measured by WST-1 analysis. Studies characterizing of the 4-OH Tam resistance of MCF7-TamR cells were performed by 17 beta-oestradiol (E2) and Annexin V/PI analysis. In addition, the expression levels of ABCC1, ABCG2 and ABCG1 were detected by RT-PCR, any changes in morphological of each resistance group were observed at the end of each month and compared with parental MCF7 cells. Consequently, exposure time and concentration can affect the degree of resistance to 4-OH Tam; thus, dose and treatment duration should be chosen according to the desired degree of resistance. This work presents a novel procedure for the generation of MCF7-TamR cells, thus enabling the identification and characterization of MCF7-TamR cells

Synthetically Lethal BMN 673 (Talazoparib) Loaded Solid Lipid Nanoparticles for BRCA1 Mutant Triple Negative Breast Cancer

PurposeThe purpose of the study was to produce BMN 673 loaded solid lipid nanoparticles (SLNs) to improve its therapeutic index, to minimize toxicity and to overcome homologous recombination (HR)-mediated resistance.MethodsFirstly, BMN 673-SLNs were characterized using Nano Zeta Sizer. After treatment with different concentrations of BMN 673 and BMN 673-SLNs, cell viability of HCC1937((BRCA1-/-)), HCC1937-R (BMN 673-resistant) TNBC and MCF-10A normal human mammary breast epithelial cell line was analyzed by WST-1 assay. In an attempt to assess the therapeutic synthetic lethality efficacy of SLNs formulation, cell cycle arrest, DNA damage, mRNA expression levels of PARP1, H2AFX, RAD51 and BRCA1 gene were investigated. Then, PARP, ?H2AX, RAD51 and BRCA1 protein expression and nuclear localization were analyzed by western blot and immunofluorescence analysis.ResultsWhen compared with BMN 673, BMN 673-SLNs showed remarkably a decrease in HCC1937 and HCC1937-R cells with less damage to MCF-10A cells. BMN 673-SLNs significantly induced toxicity through double-stranded DNA breaks, G2/M cell cycle arrest and PARP cleavage in TNBC cells. Additionally, BMN 673-resistance was mediated by miR-107, miR-193b and miR-1255b targeting BRCA1 and RAD51 in HCC1937 and HCC1937-R cells. However, BMN 673-SLNs treatment could overcome HR-mediated resistance in TNBC cells.ConclusionsAs a result, our findings suggest that SLNs formulation strongly provides a synthetic lethal therapeutic potential in BRCA1 mutated sensitive and resistant TNBC cells

E-Health and Bioengineering Conference

We mentioned the importance of clinical sequence analysis in risk determination, diagnostic and therapeutic process of familial breast cancer and we also summarized next generation sequencing applications in this cancer type. In conclusion, BRCA1/2 genes mutations are associated with an increasing the risk of particularly familial breast cancer. However, sequencing of moderate penetrance genes and/or whole exome could also fill large knowledge gaps in explaining genetic predisposition of breast cancer

E-Health and Bioengineering Conference

The anti-estrogen tamoxifen (Tam) is the most preferred option for patients with estrogen-receptor (ER)-positive breast cancer. However, multi-drug resistance (MDR) is a considerable clinical problem in the successful chemotherapeutic treatment. Members of the ATP-binding cassette (ABC) transporter family proteins play an important role in acquired drug resistance. Many studies have focused primarily on the clinical significance of P-gp (MDR1), BCRP and MRP1 members belong to ABC transporter superfamily on anticancer-drug resistance. Consequently, several strategies have been improved to overcome drug resistance. Nanoparticle drug delivery systems provide an increase in the intracellular concentration of the drugs as well as a reduction in toxicity of free-drug on healthy cells thanks to unique physical and biological properties. Solid lipid nanoparticles (SLNs) have been improved as an alternative colloidal drug delivery systems due to successful incorporation of both hydrophilic and hydrophobic compounds and their related benefits (controlled drug release, high entrapment efficiency and small size etc.) For this purpose, the aim of this study was to discuss the role of Tam-loaded solid lipid nanoparticles (SLNs) to overcome MDR and determine the ability of Tam-SLNs to induce apoptosis

BMN 673 (talazoparib): A potent PARP inhibitor for triple negative breast cancer with different genetic profile

The objective of the present study was to elucidate the effect of BMN 673 (talozoparib) on BRCA1 mutant (HCC1937) and wild-type (MDA-MB-231) triple negative breast cancer (TNBC). The in vitro cytotoxicity results indicated that BMN 673 had considerable inhibitory effects on HCC1937 and MDA-MB-231 cell lines by inducing apoptosis, multicaspase activity, G2/M arrest, and altering the expression levels of apoptosis-related genes (P < 0.01). Additionally, BMN 673 indicated no toxicity on MCF-10A control cells until a certain concentration and incubation time. However, BMN 673, a novel and selective poly ADP ribose polymerase inhibitor, was more potent in TNBC cells bearing BRCA1 mutant than those with wild-type BRCA1. In conclusion, our study, for the first time, demonstrated a molecular mechanism of the induction of apoptosis by BMN 673 in TNBC with different genetic profile. However, further investigations regarding the exact molecular mechanisms underlying BMN 673-inducing apoptotic death and gene-cell line associations are required