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S1. Details on extraction and isolation of compounds (1, 4, 6, 7, 8) from Synsepalum zenkeri; S2. Further details on tested compounds. (DOCX 24 kb)

Phages are highlyimmunogenic molecules capable of inducing both humoral and cellular immuneresponse and have been used as potential vaccine delivery vehicles. In thisstudy, the full-length Hepatitis B surface antigen (HBsAg) was expressed as HBsAg-pIIIfusion protein on M13 phage and the efficiacy of recombinant phage was studiedin BALB/cJ mice. It was shown that, immunization of mice with the highest doseof recombinant phage (1012) generated an increased antibody responsecompared to wild type phage immunization. After having a promising result inBALB/cJ mice, recombinant phage was tested in Hepatitis B virus (HBV)transgenic mice and hybrid mice (C57BL/6JXBALB/c) andits effect was also compared with the well known adjuvants such as alum andmineral oil adjuvants. Our results have shown that immunization of BALB/cJmice, HBV transgenic mice and hybrid mice with recombinant phage elicited an antigen-specificantibody response. The results also indicated that very strong adjuvants/carriersshould be used to induce immune response in HBV carriers., Fajlar organizmadahumoral ve hücresel immün yanıtıuyandırabilen yüksek immünojenik özelliğe sahip moleküllerdir ve potansiyel aşıtaşıyıcısı olarak kullanılmaktadırlar. Bu çalışmada Hepatit B yüzey antijeni (HBsAg)tam molekül olarak M13 faj yüzeyinde füzyon protein olarak ekprese edilmiş verekombinant fajın antijenik etkisi BALB/cJ farelerinde incelenmiştir. Farelerinyüksek dozda (1012) rekombinant faj ile immünize edilmesinin normalfaja kıyasla daha yüksek antikor yanıtına neden olduğu gösterilmiştir. BALB/cJfarelerinde elde edilen olumlu yanıt sonrası rekombinant fajın etkinliğiHepatit B virüsü (HBV) transgenik farelerdeve hibrid farelerde (C57BL/6JXBALB/c) incelenmiş ve ayrıca etkisi alüminyumadjuvantı ve mineral yağ adjuvantı gibi yaygın kullanımı olan adjuvantlarlakarşılaştırılmıştır. Çalışmanın sonuçları BALB/cJ farelerin, HBV transgenik farelerinve hibrid farelerin rekombinant faj ile immünizasyonunun antijene özgü antikoryanıtı oluşturduğunu göstermiştir. Sonuçlar ayrıca HBV taşıyıcılarında immünyanıt oluşturmak için güçlü adjuvantların kullanılması gerektiğini de işaretetmektedir.

Additional file 4. 1H and 13C NMR spectra of mundulea lactone 4 as well as all the 2D NMR data justifying the revision of the NMR assignment of this compound as shown in Table 2.

WOS: 000325511300002, PubMed ID: 24266102, The aims of this study are the investigation of the effects of fibronectin and type IV collagen extracellular matrix proteins and the role of caspase-3 and -9 on cis-platin induced U2-OS apoptosis were studied. First the cytotoxic effects of cis-platin on cell system were investigated by colorimetric method and than morphological and ELISA analysis were used for determination of cell apoptosis when induced with dis-platin. In addition, after adhering the cells to fibronection or type IV collagen proteins, the apoptotic rate and the effects of caspase-3 and -9 were also investigated by ELISA in presence of specific inhibitors. U2-OS cells showed 20% cytotoxicity after treatment with 2.4 mu M of cis-platin for 48 h. Morphological and the numerical data showed that cis-platin was able to induced apoptosis on cells as a dose-dependent manner. Caspase-3 and -9 inhibitors inhibited cis-platin-induced apoptosis in U2-OS cells, respectively. The binding of cells to 10 mu g/mL of fibronectin but not type IV collagen enhanced the apoptosis about 2.5 fold that effects inhibited with caspase-3 inhibitor. The baspase-3 and -9 are involved in the apoptotic signals induced by cis-platin in U2-OS. The binding to fibronectin, but not type IV collagen enhanced the apoptotic response of U2-OS and fibronectin-dependent apoptosis was activated by caspase-3. These finding might be useful for patients to fight against osteosarcoma., Anadolu University of Project Department [030326], This work was supported by grants from the Anadolu University of Project Department (No. 030326).

Dendritic cell (DC) vaccines are a promising and potent therapeutic tool for chronic diseases, autoimmune diseases, and cancer because of the unique ability of DCs to stimulate T cells. The challenge of DC vaccines is to find an effective form for antigen presentation. Although pure antigens, antigen complexes, plasmids, and mRNA have been used in different studies, no proper application to overcome this problem has been found yet. In this study, we investigated the eligibility of a commercial hepatitis B virus (HBV) vaccine or a vaccine–monoclonal antibody complex for antigen loading of DCs for a therapeutic purpose. DCs were derived from the bone marrow of transgenic hepatitis B (HBV-tg) mice using a granulocyte macrophage-colony stimulating factor and interleukin-4, and then loaded with a commercial HBV vaccine (containing hepatitis B virus surface antigens and aluminum hydroxide adjuvant) or a vaccine–antibody complex. HBV-tg mice were immunized with the vaccine and vaccine–antibody loaded DCs. Optimum HBV vaccine concentration and loading time were determined by 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium (WST- 1) methods. Therapeutic effects of vaccine–antibody loaded DCs were determined by the evaluation of antibody response and hepatitis B surface expression levels in HBV-tg mice. Our results showed that commercial HBV vaccine loaded DCs induced humoral response in HBV-tg mice but had no effect on cellular immunity.

WOS: 000252713900022, PubMed ID: 17726729, Naturally occurring compounds can have protective effects towards mutagens and carcinogens as shown by numerous studies. In the present study, the genotoxic/antigenotoxic activities of the extract from lichen Cetraria aculeata (Schreb.) Fr., which has been identified as an antibacterial agent in former studies, were investigated against known mutagens such as 4-nitro-o-phenylenediamine (4-NPD) and 2-aminofluorene (2-AF) in TA98 and TA100 strains of Salmonella typhimurium in the presence or absence of metabolic activity. Further genotoxicity/ antigenotoxicity of the extract against mitomycin C for micronucleus formation was also evaluated in human lymphocytes. The cytotoxic effects of the extract on mammalian cells were investigated in three different cell line types by MTT assay. The results obtained show that the extract of C. aculeata has a significant antigenotoxic activity in bacterial systems, but not in mammalian cells and cytotoxic activity in some mammalian cancer cells. However, it was not mutagenic in all systems. The findings suggest that the lichen extract may have a possible therapeutic potential and therefore this must be further investigated by other multiple in vitro bioassays for the development of therapeutic agents. Copyright (c) 2007 John Wiley & Sons, Ltd.

32nd Congress of the Federation-of-European-Biochemical-Societies (FEBS) -- JUL 07-12, 2007 -- Vienna, AUSTRIA, WOS: 000253283801010, …, Federat European Biochem Soc

WOS: 000243897600014, Five 2-aryl-substituted-benzothiazoles were synthesized from o-amino-thiophenol and their structural assignments were made by H-1-NMR. Mutagenicily of derivatives was tested by Salmonella microsome assay using TA98 and TA100.2-m-tolil-benzothiazole and 2-p-tolil-benzothiazole whose phenyl rings possess methyl group showed significant mutagenic activity in T100, whereas they were cytotoxic for TA98. 2-pyridin-2-yl-benzothiazole was weakly mutagenic. Tested compounds may show different genotoxicity regarding to electronic properly of alkyl substituent which is attached to phenyl ring at the second position of benzothiazole.

10th International Congress of Toxicology -- JUL 11-15, 2004 -- Tampere, FINLAND, WOS: 000222348900434, …

10th International Congress of Toxicology -- JUL 11-15, 2004 -- Tampere, FINLAND, WOS: 000222348900574, …

WOS: 000180054100007, By RT-PCR technique mRNA levels of platelet-derived growth factor type-beta receptor (PDGF-beta-R) and MyoD1 in a myoblast cell line carrying human N-ras oncogene have been investigated. After dexamethasone treatment an activation of N-ras and inhibition of the PDGF-beta-R and MyoD1 genes transcription was observed. Those results suggested that cells transformed with the N-ras oncogene fail to respond to PDGF-BB due to the absence of PDGF-beta-R mRNA.

WOS: 000169888500300, …

The essential oil Origanum onites L. and its phenolic constituent carvacrol were examined for their cytotoxic and apoptotic effects in a human hepatocellular carcinoma cells Hep-G2. WST-1 and neutral red uptake assays were performed to determine the inhibitory effects of the oil and carvacrol on the growth of the cells. Possible induction of apoptosis by Origanum oil and carvacrol was further investigated by acridine orange/ethidium bromide (AO/EB) staining. Results showed that the Origanum oil and carvacrol was significantly cytotoxic and induced apoptosis in Hep-G2 cells. IC₅₀ value of essential oil and carvacrol was found about 0,009% (v/v) and 500 μM, respectively. After incubation of the cells with Origanum oil and carvacrol, characteristics of apoptotic morphology such as chromatin condensation, shrinkage of the cells and cytoplasmic blebbing was observed. In conclusion, both essential oil and its major constituent carvacrol significantly exhibited cytotoxic and apoptotic activities in hepatocellular carcinoma cells, indicating its potential for use as an anticancer agent., Origanum onites L. uçucu yağı ve onun fenolik bileşeni olan karvakrolün insan kanserli karaciğer hücreleri Hep-G2 üzerindeki sitotoksik ve apoptotik etkileri çalışılmıştır. Hücrelerin çoğalması üzerine origanum uçucu yağının ve karvakrolün engelleyici etkilerini belirlemek için WST-1 ve nötral kırmızısı alım yöntemleri uygulanmıştır. Origanum yağı ve karvakrolün olası apoptotik etkisi akridinoranj/etidyumbromür (AO/EB) boyama yöntemiyle araştırılmıştır. Sonuçlar değerlendirildiğinde, Origanum yağı ve karvakrolün Hep-G2 hücrelerinde anlamlı derecede sitotoksik etkiye sahip olduğu ve bu hücrelerde apoptozu uyardığı gözlenmiştir. Uçucu yağın IC₅₀ değeri % 0.009, karvakrolün 500 mM olarak belirlenmiştir. Origanum uçucu yağı ve karvakrol ile inkübasyonun ardından hücrelerin kromatin yoğunlaşması, hücre büzülmesi, sitoplazmik keselerin oluşumu gibi karakteristik apop toz morfolojisi gösterdiği belirlenmiştir. Sonuç olarak, hem Origanum uçucu yağı hem de onun temel bileşeni olan karvakrol, insan kanserli karaciğer hücrelerinde anlamlı bir sitotoksik etki ve apoptotik aktivite sergilemiştir ve bu özelliklerinden dolayı, her ikisi de antikanser ajan olarak kullanılabilme potansiyeline sahiptir.

Akrepler çok sayıda biyoaktif bileşen içeren venomlarmı avlanma ya da savunma amaçlı kullanan arahnid grubundan canlılardır. Türkiye’de ölümcül zehirlenmelere neden olan iki tür Leiurus quen-questriatus ve Androctonus crassicauda olup bunlar Buthidae familyasına aittir. Bu çalışmada, A. crassicauda venomu içerisinde bulunan peptid bileşenlerin saflaştırılması ve sitotoksik etkileri araştırılmıştır. Bu amaçla, Şanlıurfa ilinden toplanan akrepler laboratuar koşullarında beslenmiş ve venomları elektrostimülasyon yöntemiyle sağılmıştır. Venomun içerdiği peptid toksinler Yüksek Performans Sıvı Kromatografisi (HPLC) ile saflaştırılmıştır. Peptidlerin hücre canlılığına etkisi BC3H1 fare beyin tümörü hücrelerinde tetrazolyum tuzu testi (MTT) ile araştırılmış ve daha sonra etkin pep-tidlerin kütle spektrometresinde molekül ağırlıkları belirlenmiştir. HPLC analizi ile venomda toplam 15 adet majör fraksiyon gözlenmiştir. Bunlardan Ac8, Ac9 ve Ac10 olarak adlandırılan peptidlerin BC3H1 hücreleri için sitotoksik olduğu bulunmuş ve Ac8 için IC50 değeri 47 µg/ml, Ac9 için 72 µg/ml ve Ac10 için 40 µg/ml olarak belirlenmiştir. Moleküler ağırlıkları ise Ac8 için 16356 Da, Ac9 için 7422 Da ve Ac10 için 16328 Da olarak belirlenmiştir. Bu çalışma ile ilk kez, A. crassicauda ham venomundan sitotoksik etkili yeni peptidler saflaştırılmıştır. Bu peptidler anti-tümör potansiyele sahip olabilirler ve bu nedenle çalışılmalarının yararlı olacağı düşünülmektedir., Scorpions are arachnids containing many bioactive constituents which they use it for hunting or defense. Leiurus quinquestriatus and Androctonus crassicauda which causes lethal poisoning in Turkey are two species belonging to Buthidae family. In this study, purification and cytotoxicity of peptide constituents of A. crassicauda venom was investigated. For this purpose, scorpions collected from Şanlıurfa province were housed in the laboratory condition and their venoms were collected by electro stimulation method. Peptide toxins of the venom were isolated by High Performance Liquid Chromatograph (HPLC). Effects of the peptides on the viability of BC3H1 mouse brain tumor cells were examined by the assay of tetrazolium salt (MTT) and then the molecular weight of the active molecules were determined by mass spectrometer. Total 15 major fractions were observed by HPLC analysis. Peptides named as Ac8, Ac9 and Ac10 were found to be cytotoxic for BC3H1 cells and IC50 value was 47 µg/ml for Ac8, 72 µg/ml for Ac9 and 40 µg/ml for Ac10. Molecular weights of the peptides were also determined as 16356 Da for Ac8, 7422 Da for Ac9 and 16328 Da for Ac10. By the present study, the new peptides with cytotoxic activity were isolated from the crude venom of A. crassicauda the first time. We suggest that these peptides may have a potential as antitumor substances and therefore it may be useful to study on them.

Pyridine as a nitrogen-containing heterocyclic substance and its derivatives exist in both surface and ground water because of its use in a variety of food, medicinal and chemical industrial products. In the present work, cytotoxic effect of pyridine has been investigated for the first time in a plant, on root-tip growth and the meristem cells of onion Allium cepa L. . Different concentrations of pyridine significantly affected the growth and morphology of onion roots causing shorter, thicker and gelatin like structures compared to the control. An increase in the pyridine doses caused an increase in mitosis abnormalities in the squashes prepared from the cells of root-tip meristem whereas the mitotic index was diminished. Mitosis abnormalities have been observed generally as anaphase bridges, stickiness, anaphasetelophase aberrations and c-metaphase. As a result, pyridine at low doses has been significantly cytotoxic on the meristem cells of onion root-tip. Thus, we suggest that toxic effects of pyridine may be investigated and the mechanisms may be evaluated in different systems for human health, Azot-içeren heterosiklik bir madde olan piridin ve türevleri besin, tıbbi olarak ya da kimya sanayi gibi endüstriyel aktiviteler sonucu oluşmakta ve hem yüzey hem de yer altı sularında yaygın olarak bulunmaktadır. Bu çalışmada, piridinin soğan Allium cepa L. köklenmesi ve kök-ucu meristem hücreleri üzerindeki genotoksik etkisi ilk kez araştırılmıştır. Farklı dozlardaki piridin soğan köklerinin büyümesini ve morfolojisini doza bağlı olarak önemli derecede etkilemiş, kontrole göre daha kısa, kalın ve jelimsi yapılar olarak gelişmesine neden olmuştur. Kök ucu meristem hücrelerinden hazırlanan ezme-preparatlarda, piridin dozlarındaki artış mitotik indeksin düşmesine neden olurken, mitoz anomalilerinde artışa neden olduğu gözlenmiştir. Mitoz anomalileri genellikle anafaz köprüsü, yapışkanlık, bozulmuş anafaz-telofaz ve c-metafaz oluşması şeklinde gözlenmiştir. Sonuç olarak, düşük dozlardaki piridin soğan kök-ucu meristem hücreleri üzerinde önemli derecede sitotoksik olmuştur. Bundan dolayı, insanların sıklıkla maruz kaldığı piridinin toksik etkisi farklı sistemlerde de araştırılmalı ve etki mekanizması aydınlatılmalıdır

WOS: 000423839200002, PubMed ID: 29378558, Background: Despite the remarkable progress in cancer therapy in recent years, this disease still remains a serious public health concern. The use of natural products has been and continues to be one of the most effective ways to fight malignancies. The cytotoxicity of 14 compounds from African medicinal plants was evaluated in four human carcinoma cell lines and normal fibroblasts. The tested samples included: beta-spinasterol (1), friedelanone (2), 16 beta-hydroxylupeol (3), beta-amyrin acetate (4), lupeol acetate (5), sequoyitol (6), rhamnitrin (7), europetin 3-O-rhamnoside (8), thonningiol (9), glyasperin F (10), seputhecarpan B (11), seputhecarpan C (12), seputhecarpan D (13) and rheediaxanthone A (14). Methods: The neutral red uptake (NR) assay was used to evaluate the cytotoxicity of samples; caspase-Glo assay, flow cytometry for cell cycle analysis and mitochondrial membrane potential (MMP) as well as spectrophotometry to measure levels of reactive oxygen species (ROS) were performed to detect the mode of action of compounds 9 and 13 in MCF-7 breast adenocarcinoma cells. Results: Compounds 3, 9-13 displayed cytotoxic effects against the four tested cancer cell lines with IC50 values below 85 mu M. Compounds 9 and 13 had IC50 values below 10 mu M in 4/4 and 3/4 tested cell lines respectively. The IC50 values varied from 0.36 mu M (against MCF7 cells) to 5.65 mu M (towards colon carcinoma DLD-1 cells) for 9, from 9.78 mu M (against MCF7 cells) to 67.68 mu M (against HepG2 cells) for 13 and 0.18 mu M (towards HepG2 cells) to 72 mu M (towards Caco-2 cells) for the reference drug, doxorubicin. Compounds 9 and 13 induced cell cycle arrest in Go/G1 whilst doxorubicin induced arrest in G2/M. The two molecules (9 and 13) also induced apoptosis in MCF-7 cells through activation of caspases 3/7 and 9 as well as enhanced ROS production. Conclusion: Compounds 9 and 13 are good cytotoxic phytochemicals that should be explored more in future to develop a cytotoxic drug to fight human carcinoma., Scientific and Technological Research Counsel of Turkey (TUBITAK); Scientific Research Projects Commission of Anadolu University, Eskisehir, Turkey [1507F563]; International Science Programme, Uppsala University, Sweden through the Network for Analytical and Bioassay Services in Africa (NABSA) at the University of Botswana [(ISP)-KEN-02], The study was funded by the Scientific and Technological Research Counsel of Turkey (TUBITAK) and to Scientific Research Projects Commission of Anadolu University, Eskisehir, Turkey (funding grant 1507F563). A grant for part of this work was also provided by International Science Programme, Uppsala University, Sweden (ISP)-KEN-02 project through the Network for Analytical and Bioassay Services in Africa (NABSA) at the University of Botswana.

WOS: 000466854400002, PubMed ID: 29281926, Paraben is a phenolic derivative of benzoic acid extensively used as preservatives in food, pharmaceutical, and cosmetic industries due to its antimicrobial characteristics. The objective of this study was to evaluate the in vitro genotoxic effects of paraben in human lymphocyte cultures. Cells were analyzed by cytokinesis-block micronucleus (CBMN), chromosome aberration (CA), sister chromatid exchange (SCE), and comet tests. For CBMN, CA, and SCE assays, the human lymphocytes were isolated from healthy donors and incubated with 500, 250, 100, and 50 mu g/mL of paraben for 24 and 48 h, and for comet assay, cells were exposed to 1000, 750, 500, and 250 mu g/mL of paraben for an hour. Results showed that numbers of MN and SCEs were not significant in the cells exposed to paraben when compared to the solvent control. However, 500 and 250 mu g/mL of paraben induced the CA after 24 h. Also, we observed a significant decrease in the cytokinesis-block proliferation index in cells exposed 250-500 mu g/mL paraben for 24 h, and 100, 250, and 500 mu g/mL for 48 h. The mitotic index was also decreased at all concentrations and periods. However, the proliferation index was statistically decreased at all concentrations after 48 h treatments. Only the highest concentration of paraben caused DNA migration (mean tail length) in human lymphocytes analyzed by Comet assay. Taken together, results indicated that paraben had cytotoxic effects and caused genotoxicity by affecting directly chromosomes and DNA in human lymphocyte cells in vitro, and may have genotoxic potential for human., Anadolu University Research Fund [1204F066], This study was supported by the Anadolu University Research Fund (Project No: 1204F066).

WOS: 000462182300013, Objective: The aim of this study is to investigate in vitro effects of an antihelminthic drug niclosamide on human ovarian carcinoma cell line OVCAR3. Methods: MTT assay was applied to investigate the cytotoxic effects of niclosamide on the cells. beta-catenin levels in the cells were analyzed by immunocytochemistry, in order to assess the potency of niclosamide on Wnt/beta-catenin signaling pathway that function in cell proliferation. The effects of the drug on apoptosis were detected by TUNEL method. All the assays were also performed for chemotherapy agent 5-fluorouracil (5-FU) and anticancer effects of these two drugs were compared. Results: It was found that niclosamide at 1 mu M and 2 mu M concentrations reduced cell viability, whereas 5-FU showed its significant proliferation inhibitory effect at higher concentrations. Niclosamide led to an increase in apoptosis while this effect was weaker compared with 5-FU. Niclosamide treatment decreased beta-catenin staining in the cells significantly but 5-FU did not affect beta-catenin levels. Conclusion: The results indicate that niclosamide induces apoptosis and suppresses cell proliferation by inhibiting Wnt/beta-catenin signaling pathway in OVCAR-3 cells. In conclusion, these findings warrant further evaluation of niclosamide as a promising therapy for ovarian cancer.

Tez (doktora) - Anadolu Üniversitesi, Anadolu Üniversitesi, Fen Bilimleri Enstitüsü, Biyoloji Anabilim Dalı, Kayıt no: 480268, Metastazik kolon kanseri tedavisinde yaygın olarak kullanılan ön ilaç irinotekanın aktif metaboliti, UGT1A1 tarafından metabolize edilen SN-38’dir. Genetik ve epigenetik faktörlerin ilaç yanıtı üzerindeki etkisi ilaçların akılcı kullanımını öne çıkarmaktadır. Bu çalışmada, UGT1A1 ifadesini etkileyebilecek genetik ve epigenetik faktörlerin irinotekan ilaç yanıtındaki çıktıları hakkında öngörüsel veri sahibi olunması hedeflenmiştir. Çalışmamızda irinotekan tedavisi alan kolon kanserli 48 hasta ile çalışılmıştır. En sık diyare, mukozit ve nötropeni olmak üzere %66,6'sında toksik etki görülmüştür. UGT1A1 1*/1* yaygın yabanıl genotip %72,98, UGT1A1 28*/28* çekinik genotip %6,5 bulunmuştur. Epigenetik analizleri için 9 hastaya ait kan örneği ve 8 hastaya ait 13 adet kolon doku kesiti ile çalışılmıştır. UGT1A1 transkripsiyon başlama noktasının yukarısındaki 3., 4., 28., 29. ve 30. CpG dinükleotidlerinin metilasyon profili incelenmiştir. Promotor bölgede -4 CpG ve enhancerdaki -28 CpG ve -29 CpG dinükleotidlerinin metillendiği saptanmıştır. Kan örneklerinde doku örneklerine kıyasla tüm bölgelerde yüksek metilasyon profiline rastlanmıştır. UGT1A1 genotipleri, hasta yaşam biçimi, demografik faktörler, kombine ilaç kullanımları ve UGT1A1 CpG dinükleotid metilasyonları çoklu veya ikili lojistik regresyon modeli ile incelenmiştir. Bu faktörler arasında anlamlı bir ilişki bulunamamıştır. Bu çalışma kan örneklerinde UGT1A1 için metilasyon paterninin çalışıldığı ilk çalışma olup, UGT1A1'in dokuya özgü metillenerek epigenetik regülasyonunun olabileceğini göstermiştir. Ayrıca Türkiye'de irinotekan farmakoepigenetiği ile yapılan ilk araştırmadır.

Hepatosellülar karsinoma (HSK) kanser ilişkili ölümler sıralamasında dünyada ikincidir. Epitel-mezenkimal geçiş (EMT) metastaz başlaması için önemli bir programdır. Buna rağmen, pek çok kanser hücresi bütün epitel karakterleri tamamen kaybetmeden yada bütün mezenkim karakterleri tamamen kazanmadan kısmi EMT programıyla metastaz yapabildiği de bilinmektedir. Bu tez çalışmasında, tam ve kısmi EMT'nin benzeyen ve ayrışan yönlerini tespit etmek için EMT ile ilişkili transkripsiyon faktörü olan Slug'ın iki farklı HSK hücre hattında aşırı ifadesi sağlandı. Sonra hücreler EMT fenotipi, kemodirenç ve kök hücre özellikleri bakımından analiz edildi. Aşırı Slug ifadesinin HSK'da hem tam hem de kısmi EMT'yi tetiklediği gösterildi. Tam EMT, E-cadherin'in azalması, ZEB2 mRNA'sının ise artması ile karakterize edildi. Kısmi EMT ise E-cadherin'in artması ve vimentin ve ZEB2 mRNA'sının azalması ile karakterize edildi. ABCB1 azalması ve ABCG2 artması ile ortaya çıkan Slug aracılı kemodirenç gelişimi ilginç bir şekilde hem tam hemde kısmi EMT'de ortak olarak olarak gözlendi. Buna ilave olarak kök hücre karakteri olan CD133 mRNA'sının artması her iki EMT için ortak olan diğer bir özellikti. Bunların yanında, kısmi EMT fenotipi gösteren HSK hücrelerinden izole edilen koşullu medyumda, translasyon sonrası değiştirilmiş fibronektin1 ve kollajen tip II α 1 ile değiştirilmemiş fibrinojen gama zincirinin artması tespit edildi. Böylece bu proteinlerin kemodirenç ve kısmi EMT biyoişaretleri olarak kullanılabileceği ileri sürüldü. Aynı şekilde koşullu medyumda, hsa-miR-296-3p, hsa-miR-101-3p ve hsa-miR-885-5p miRNA miktarlarının artması tespit edildi. Böylece bu mikroRNA'ların da HSK'da kemodirenç ve kısmi EMT biyoişaretleri olarak kullanılabileceği ileri sürüldü. Hepatocellular carcinoma (HCC) is the second-leading cause of cancer-related deaths worldwide. Complete EMT has long been considered as a crucial step for initiating metastasis. However, it is now known that many carcinoma cells can metastasize with incomplete loss of epithelial traits or incomplete gain of mesenchymal phenotype, which is named as partial EMT. Here, in order to determine the similarities and differences between complete and partial EMT phenotype in HCC, the EMT associated transcription factor Slug was overexpressed in different HCC cell lines and the cells were compared for their EMT phenotype, chemoresistance and stemness. Conditioned medium was also collected from a HCC cell line overexpressing Slug and was analyzed for markers of chemoresistance and partial EMT. We observed that Slug overexpression resulted in the induction of both complete and partial EMT in different HSK cell lines. Complete EMT was characterized by the down regulation of E-cadherin and up-regulation of ZEB2 mRNA. Partial EMT was characterized by the up-regulation of E-cadherin and down-regulation of vimentin and ZEB2 mRNA. Interestingly, Slug mediated chemoresistance through the down-regulation of ABCB1 and up-regulation of ABCG2 mRNAs as well as expression of CD133, a stemness marker was similar in both complete and partial EMT phenotype. On the other hand, partial EMT was associated with enhanced secretion of post translationally modified fibronectin1, collagen type II alpha 1 and native fibrinogen gamma chain upregulations in the conditioned medium of HSK cells. These proteins can therefore be considered as potential biomarkers of chemoresistance and partial EMT in HSK. Similarly, in the conditioned medium, hsa-miR-296-3p, hsa-miR-101-3p, hsa-miR-885-5p, miRNAs were found to be up regulated respectively. It has been suggested that these microRNAs can also be used as chemoresistance and partial EMT biomarkers in HCC. 125