Identification of intracellular pathways through which TGF-β1 upregulates URG-4/URGCP gene expression in hepatoma cells

Tokay, Esra; Köçkar, Feray

doi:10.1016/j.lfs.2015.12.010

Cited by 19 publications

(7 citation statements)

References 35 publications

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“…The data represented using Graphpad Prism 7 programme. IC50 values were calculated using Origin Pro 8.5 programme [13].…”

Section: Mtt Analysesmentioning

confidence: 99%

Determination of Cytotoxic effect and Expression analyses of Apoptotic and Autophagic related genes in Thymoquinone-treated Colon Cancer Cells

Tokay

2020

Sakarya University Journal of Science

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Thymoquinone (Tq; 2-isopropyl-5-methyl-1,4 benzoquinone) is the main ingredient that presents in the volatile oil of Nigella sativa (black seed). Recently, it has been reported that, Tq was inhibited cell proliferation of many cancer cell lines including ovarian, lung, breast, osteosarcoma, pancreatic, fibrosarcoma, lung cancer, squamous cell carcinoma and myeloblastic leukemias. In this study, the anti-cancer effect of Tq was investigated in human Colon Cancer Cells (HT-29) with MTT assay. Positive control was achieved with 5-fluorouracil (5-FU). Also, the invasion capability of HT-29 cells was determined in presence of Tq and 5-FU with scratch analyses. In addition, the autophagic and apoptotic effect of Tq were investigated by qRT-PCR method to find out the pathway of anti-proliferative effect in Tqtreated HT-29 cells. As a result, Tq was inhibited the cell proliferation of HT-29 cells in time dependent manner, with 118 µM for 24 h and 84 µM for 48 h, respectively. A concentration of 150 µM of Tq was able to significantly reduced the invasiveness of HT-29 cells approximately 8 fold compared with not treated cells at 6h. ATG-12, ATG-7 and LC3-II were significantly downregulated in presence of Tq. Conversely, the expression of all the autophagy genes were downregulated in presence of 5-FU. Moreover, pro-apoptotic gene Bax was significantly upregulated nearly 15 fold whereas the expressions of Bcl-2 and Bcl-XL (prosurvival genes) were decreased in presence of Tq. On the other hand, in 5-FU treated HT-29 cells, the expression of Bax, Bcl-2 and Bcl-XL genes and autophagy related genes were significantly downregulated.

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“…The data represented using Graphpad Prism 7 programme. IC50 values were calculated using Origin Pro 8.5 programme [13].…”

Section: Mtt Analysesmentioning

confidence: 99%

Determination of Cytotoxic effect and Expression analyses of Apoptotic and Autophagic related genes in Thymoquinone-treated Colon Cancer Cells

Tokay

2020

Sakarya University Journal of Science

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“…The mean OD value of the control groups was subtracted from the OD value of each cytokine treated groups for all incubation times. Experiments were performed for three times [35].…”

Section: Cell Culture and Mtt Assaymentioning

confidence: 99%

Effect of Angiogenesis Related Growth Factors VEGF-a and FGF-1 on Osteosarcoma Cell Proliferation

Alper¹

2020

Adıyaman University Journal of Science

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Osteosarcoma is the most common bone tumor in children and adolescents. Alterations in the expression of some genes, cytokines and growth factors are responsible for the development of the malignant phenotype in osteosarcoma. Some members of the VEGF and FGF families have been associated with poor prognosis and the metastasis in various tumor types including osteosarcoma. Among the members of the family, information about effects of VEGF-a and FGF-1 on osteosarcoma cell proliferation is limited.In the present study, it was aimed to elucidate the effects of VEGF-a and FGF-1 on osteosarcoma cell proliferation. Saos-2 cells, having osteoblastic features, were used for osteosarcoma model. Cells were treated 20 ng/mL VEGF-a and FGF-1 after 24 hours of serum starvation. MTT assay was applied to measure cell viability after incubation for 1-72 hours.Results indicated that VEGF-a promoted cell proliferation for all incubation times. Maximum increase was observed after 48 hours of incubation (1.7 fold) with a statistically important manner.FGF-1 led to very slight increase in Saos-2 cell proliferation. Consequently, these findings can contribute development of new therapy strategies for osteosarcoma.

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“…Cloning of the Ssap-NtrB was performed in a three-step procedure (Figure 1). Firstly, the PCR product was cloned into the pGEM-T Easy vector (Promega) with T: A cloning system [13,14]. Subsequently, transformation was carried out.…”

Section: Cloning Strategy Of Ssap-ntrb Gene Into Eukaryotic Expression Vectormentioning

confidence: 99%

“…For this purpose 3 µL of vector (1000 ng/µL), 23 µL of plasmid (780 ng/ µL), 1 µL of T4 DNA ligase Enzyme (5 U/µL) and buffer (1X) in a reaction volume of 30 µL. Ten microliters of ligation mixture were transferred into 200 µL E. coli XL1Blue (the competency at 1 x 10 6 colony-forming unit / μg DNA/100 μL cells) competent cell lines [13][14][15]. Overnight cultures were prepared from each of the resulting colonies.…”

Section: Cloning Strategy Of Ssap-ntrb Gene Into Eukaryotic Expression Vectormentioning

confidence: 99%

Ssap-NtrB prokaryotik geninin ökaryotik ekspresyon vektörü pcDNA3.1 / V5 / His B içine klonlanması

Hacıoğlu

Köçkar

2020

Balıkesir Üniversitesi Fen Bilimleri Enstitüsü Dergisi

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Suicide gene therapy has recently emerged as a method used in cancer treatments. These therapies utilized enzymes that are expressed in the cell. In this study, Staphylococcus saprophyticus supsp. saprophyticus Nitroreductase gene (Ssap-NtrB) was subcloned into the eukaryotic expression vector namely pcDNA3.1 / V5 / His B. For this purpose, Nitroreductase gene region was firstly amplified from the pET14B vector using PCR strategy and cloned into the pGEM-T-Easy vector. After this step, the Ssap-NtrB gene was restricted with KpnI/ApaI and was ligated into pcDNA3.1 / V5 / His B vector. Recombinant colonies were verified using KpnI/ApaI restriction enzymes. As a result, the Ssap-NtrB gene was cloned into pcDNA3.1/V5/His B vector and was readyfor use in suicide gene therapy in eukaryotic human cancer cells.

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Identification of intracellular pathways through which TGF-β1 upregulates URG-4/URGCP gene expression in hepatoma cells

Cited by 19 publications

References 35 publications

Determination of Cytotoxic effect and Expression analyses of Apoptotic and Autophagic related genes in Thymoquinone-treated Colon Cancer Cells

Determination of Cytotoxic effect and Expression analyses of Apoptotic and Autophagic related genes in Thymoquinone-treated Colon Cancer Cells

Effect of Angiogenesis Related Growth Factors VEGF-a and FGF-1 on Osteosarcoma Cell Proliferation

Ssap-NtrB prokaryotik geninin ökaryotik ekspresyon vektörü pcDNA3.1 / V5 / His B içine klonlanması

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