Zeynep Akbulut

Amaç: Bu çalışmada, Hypericum perforatum (HP) ekstraktının HepG2 karaciğer kanseri hücrelerinde hücre canlılığı ve p53 proteininin hücre içi lokalizasyonu üzerindeki etkilerinin araştırılması amaçlandı. Gereç ve Yöntem: HepG2 hücreleri standart kültür koşullarında çoğaltıldı. Deney grubuna 24 saat süreyle %0,8 HP ekstraktı uygulanırken kontrol grubuna herhangi bir uygulama yapılmadı. Hücre canlılığı CCK-8 testi ile değerlendirildi. p53 proteininin hücre içi dağılımı immünfloresan boyama ve konfokal mikroskopi ile incelendi. İstatistiksel analizlerde GraphPad Prism 10.0 kullanıldı ve p<0,05 anlamlı kabul edildi. Bulgular: %0,8 HP ekstraktı uygulanan HepG2 hücrelerinde kontrol grubuna kıyasla hücre canlılığında anlamlı azalma saptandı (p<0,05). Toplam p53 immünfloresan yoğunluğu açısından gruplar arasında belirgin fark gözlenmezken, konfokal mikroskopi analizleri HP uygulanan hücrelerde p53’ün sitoplazmadan çekirdeğe doğru yeniden dağılım gösterdiğini ortaya koydu. Sonuç: Hypericum perforatum ekstraktı HepG2 hücrelerinde hücre canlılığını azaltmış ve p53’ün hücre içi lokalizasyonunda değişikliğe neden olmuştur. Bulgular, HP ekstraktının hücresel stres yanıtını tetikleyebileceğini düşündürmektedir. Bununla birlikte, etkinin p53 aracılı apoptotik mekanizmalar üzerinden gerçekleşip gerçekleşmediğinin ortaya konulabilmesi için doz-cevap çalışmaları ve ileri moleküler analizlere ihtiyaç vardır. Anahtar Kelimeler: Hypericum perforatum, HepG2, hücre canlılığı, p53, karaciğer kanseri, CCK-8

Purpose: The tumor microenvironment significantly influences hepatocellular carcinoma (HCC) progression, with epithelial-mesenchymal transition (EMT) being a key driver of metastasis. This study explores the distinct roles of matrigel and type I collagen, two prominent ECM components, in modulating HepG2 liver cancer cell behavior. Materials and Methods: HepG2 cells were cultured in type I collagen and Matrigel culture media, and the expression of YAP1, Na-K ATPase, and E-cadherin were investigated using a confocal microscope. We examine their impact on proliferation and the expression of, considering the influence of cell adherent. Results: HepG2 cells showed the highest proliferation at post-confluent culture time in type 1 collagen medium. YAP1 expression was high in type 1 collagen medium at both culture times (and when they reached adherent, YAP1 localization was from the cytoplasm to the nucleus. Na-K ATPase expression was high in control cells non-adherent, while it increased in Matrigel adherent. E-cadherin showed a statistically significant decrease in type 1 collagen medium compared to control and its expression was found to be high in Matrigel cultured cells adherent. Conclusion: These findings highlight the differential effects of extracellular matrix components on HCC cell proliferation and EMT-related protein expression, suggesting potential therapeutic targets within the tumor microenvironment.

Amaç: Bu çalışmada yeni tanımlanan bir proto-onkogen olan ve vücutta çok fazla A vitamini birikmesiyle ortaya çıkan hipervitaminoz A hastalığıyla ilişkili olduğu bilinen homeobox (ISX) proteininin, Hepatoselüler karsinom (HCC) hücre hattının farklı ekstrasellüler matriks ortamlarda kültüre edilmesiyle, ISX gen ekspresyonu düzeyinde gösterilmesi, ayrıca kültür ortam faklılıklarının kanser kök hücre belirteçleri üzerindeki etkisinin değerlendirmesi amaçlanmıştır. Materyal ve Metodlar: Karaciğer karsinom hücre hattı olan HepG2 hücreler tip I kollajen ve matrigel kültür ortamlarda kültüre edilerek, ISX geninin ekspresyonu real time PCR kullanılarak, alfa-fetoprotein (AFP), kök hücre belirteçleri olan OV-6 ve Nanog proteinlerinin hücresel miktarları ve lokalizasyonlarındaki değişiklikler konfokal mikroskop ile araştırılmıştır. Bulgular: ISX gen ekspresyonu, kontrol grubuna göre Matrigel kültür ortamda 4 kat, tip I kolajen kültürde ise 2 kat daha fazla tespit edilmiştir. Oct-4 antikorunun ekspresyonu tip I kollajen kültür ortamında kontrol grubuna göre istatistiksel olarak anlamlı derecede yüksektir (p<0,05). Nanog antikoru, kontrol grubuna göre Matrigel ortamda HepG2 hücrelerinde anlamlı bir azalma gösterirken (p0,05). Sonuç: Bu çalışmadan elde edilen sonuçların karaciğer kanseri mekanizmaları ve kanser kök hücrelerinin davranışını etkileyecek tedavi seçeneklerinin geliştirilmesi konusunda yapılacak ileri çalışmalara yardımcı olabileceği düşünülmektedir. Anahtar kelimeler: Hepatoselüler karsinom, A vitamini, ISX geni, Tip1 kollajen, Matrigel

Objectives: Sorafenib is an orally administered tyrosine kinase inhibitor in hepatocellular cancer. Low sorafenib concentrations are attained during pharmacotherapy due to pharmacokinetic profile and patient inadherence. Resistance to treatment is a limitation to improving survival. Underlying mechanisms include epithelial-mesenchymal transition. The aim of the study was to evaluate epithelial-mesenchymal transition and multidrug resistance-related parameters in HepG2 cells following low-dose and short-term sorafenib treatment. Methods: Epithelial-mesenchymal transition and multidrug resistance-related markers were examined by quantitative PCR, flow cytometry, and confocal laser scanning microscopy. Results: An increase in epithelial marker E-cadherin and downregulation of mesenchymal markers Vimentin and Snail1 were detected by gene expression analysis. While P-glycoprotein expression increased, multidrug resistance protein 1, and breast cancer resistance protein mRNA levels did not alter after sorafenib treatment. The accumulation of the ABC transporter substrate rhodamine 123 in the cells increased following the treatment, corresponding to a less efficient efflux of rhodamine 123 and a possible effect on other transporters and mechanisms. Conclusions: The results indicate a protective effect of sorafenib against epithelial-mesenchymal transition and upregulation in P-glycoprotein expression, which is, however, not sufficient to cause less intracellular rhodamine 123 accumulation. The effects of low-dose and short-term sorafenib on epithelial-mesenchymal transition and multidrug resistance-related markers might contribute to enlightening new treatment strategies in hepatocellular cancer.

This study aims to evaluate humoral immune system response by measuring copy numbers of kappa-deleting recombination excision circles (KREC) gene segment from B lymphocytes in patients with lung transplantation. Between September 2015 and November 2016, a total of 11 patients (8 males, 3 females; mean age: 45.4±12.0 years; range, 23 to 59 years) who underwent lung transplantation with different primary indications were included. The copy numbers of KREC gene segment were quantified using real-time polymerase chain reaction method in peripheral blood samples collected pre- and post-transplantation. The samples of the patients were compared with the KREC l evels i n deoxyribonucleic acid extracted from blood samples of healthy children. There was no significant change in KREC levels between pre- and post-operation (p=0.594 and p=0.657), although the median values indicated that the highest increase in the KREC levels (7x105- 12x105; 85-170) was on Day 7 of transplantation. There was a positive correlation between the KREC levels (mL in blood) and lymphocytes at 24 h after transplantation (p=0.043) and between KREC copies per 106 of blood and age on Day 7. Our preliminary results suggest that KREC l evels a s an indicator of B lymphocyte production are elevated after lung transplantation. A prognostic algorithm by tracking B cell kinetics after post-transplantation for long-term follow-up can be developed following the confirmation of these preliminary results with more patient samples.

Cisplatin, the first platinum compound approved for cancer treatment, is widely used in the treatment of various cancers including hepatocellular carcinoma (HCC). HCC incidence rates rise globally. Epithelial mesenchymal transition (EMT) is implicated in cancer invasion and metastasis, which are associated with increased mortality. Cisplatin dose might influence cancer invasion and metastatic behavior of the cells. The aim of the study was to investigate the effect of low-dose cisplatin treatment on EMT- related changes in HepG2 cells. Following treatment with 4 µM cisplatin, HepG2 cells were evaluated morphologically. Gene expression of E-cadherin, Vimentin, Snail1 was assessed by quantitative PCR. Immunofluorescence analyses of NA-K ATPase were performed. Although the low-dose cisplatin treated cells exhibited a more stretched morphology, no statistical difference was detected in gene expression of E-cadherin, Vimentin, Snail1 and immunofluorescence of NA-K ATPase. Findings on low-dose cisplatin effects in HepG2 might contribute to the knowledge of antineoplastic inefficacy by further understanding the molecular mechanisms of drug action.