scholarly journals 5-fluorouracil Toxicity Mechanism Determination in Human Keratinocytes: in vitro Study on HaCaT Cell Line

2017 ◽  
Vol 118 (4) ◽  
pp. 128-138
Author(s):  
Jan Hartinger ◽  
Pavel Veselý ◽  
Martin Šíma ◽  
Irena Netíková ◽  
Eva Matoušková ◽  
...  
Keyword(s):  
Cell Line ◽  
In Vitro Study ◽  
Human Keratinocytes ◽  
Cell Types ◽  
Toxicity Mechanism ◽  
Keratinocyte Cell Line Hacat ◽  
Keratinocyte Cell ◽  
First Time ◽  
Fluorouracil Toxicity

5-fluorouracil (5-FU) and capecitabine therapy is often accompanied by palmar-plantar erythrodysesthesia (PPE) which is manifestation of 5-FU toxicity in keratinocytes. The main mechanisms of 5-FU action are thymidylate synthase (TS) inhibition which can be abrogated by thymidine and strengthened by calciumfolinate (CF) and incorporation of fluorouridinetriphosphate into RNA which can be abrogated by uridine. For proper PPE treatment 5-FU mechanism of action in keratinocytes needs to be elucidated. We used the 5-FU toxicity modulators uridine, thymidine and CF to discover the mechanism of 5-FU action in human keratinocyte cell line HaCaT. To measure the cellular viability, we used MTT test and RTCA test. CF did not augment 5-FU toxicity and 5-FU toxicity was weakened by uridine. Therefore, the primary mechanism of 5-FU toxicity in keratinocytes is 5-FU incorporation into RNA. The uridine protective effect cannot fully develop in the presence of CF. Thymidine addition to 5-FU and uridine treated cells not only prevents the toxicity-augmenting CF effect but it also prolongs the 5-FU treated cells survival in comparison to uridine only. Therefore, it can be assumed that in the presence of uridine the 5-FU toxicity mechanism is switched from RNA incorporation to TS inhibition. Although particular 5-FU toxicity mechanisms were previously described in various cell types, this is the first time when various combinations of pyrimidine nucleosides and CF were used for 5-FU toxicity mechanism elucidation in human keratinocytes. We suggest that for PPE treatment ointment containing uridine and thymidine should be further clinically tested.

scholarly journals The Selection of NFκB Inhibitors to Block Inflammation and Induce Sensitisation to FasL-Induced Apoptosis in HNSCC Cell Lines Is Critical for Their Use as a Prospective Cancer Therapy

2019 ◽  
Vol 20 (6) ◽  
pp. 1306 ◽  
Author(s):  
Mario Scheurer ◽  
Roman Brands ◽  
Mohamed El-Mesery ◽  
Stefan Hartmann ◽  
Urs Müller-Richter ◽  
...  
Keyword(s):  
Cancer Therapy ◽  
Cell Lines ◽  
In Vitro Study ◽  
Keratinocyte Cell Line Hacat ◽  
Keratinocyte Cell ◽  
Nfκb Pathway ◽  
Induced Apoptosis ◽  
Selection Of ◽  
Hnscc Cell

Inflammation is a central aspect of tumour biology and can contribute significantly to both the origination and progression of tumours. The NFκB pathway is one of the most important signal transduction pathways in inflammation and is, therefore, an excellent target for cancer therapy. In this work, we examined the influence of four NFκB inhibitors—Cortisol, MLN4924, QNZ and TPCA1—on proliferation, inflammation and sensitisation to apoptosis mediated by the death ligand FasL in the HNSCC cell lines PCI1, PCI9, PCI13, PCI52 and SCC25 and in the human dermal keratinocyte cell line HaCaT. We found that the selection of the inhibitor is critical to ensure that cells do not respond by inducing counteracting activities in the context of cancer therapy, e.g., the extreme IL-8 induction mediated by MLN4924 or FasL resistance mediated by Cortisol. However, TPCA1 was qualified by this in vitro study as an excellent therapeutic mediator in HNSCC by four positive qualities: (1) proliferation was inhibited at low μM-range concentrations; (2) TNFα-induced IL-8 secretion was blocked; (3) HNSCC cells were sensitized to TNFα-induced cell death; and (4) FasL-mediated apoptosis was not disrupted.

scholarly journals Local Treatment of Hand-Foot Syndrome with Uridine/Thymidine:In VitroAppraisal on a Human Keratinocyte Cell Line HaCaT

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
J. Hartinger ◽  
P. Veselý ◽  
E. Matoušková ◽  
S. Argalacsová ◽  
L. Petruželka ◽  
...  
Keyword(s):  
Cell Line ◽  
Local Treatment ◽  
Local Therapy ◽  
Anticancer Therapy ◽  
Protective Activity ◽  
Human Keratinocyte ◽  
Human Keratinocyte Cell Line ◽  
Keratinocyte Cell Line Hacat ◽  
Keratinocyte Cell

5-fluorouracil (5-FU) is one of the most commonly used antineoplastic drugs in the anticancer therapy. The hand-foot (HF) syndrome (palmar-plantar erythrodysesthesia) is an adverse effect frequently related to long-term i.v. administration of 5-FU or its orally applicable prodrug capecitabine. Its severity can even lead to interruption of the otherwise effective anticancer therapy. Tentative practice in some clinics has shown that topical application of 10% uridine ointment is beneficial for calming down the HF syndrome. This study is focused on verifying the alleged protective activity of uridine in thein vitromodel of cultured human keratinocyte cell line HaCaT. We also tested the protective effects of thymidine alone or uridine-thymidine combination. The cellular viability time progression was measured in order to evaluate the effect of protective agents by three different types of cytopathogenicity tests—NTCA test (non-destructive test of cellular activity), modified MTT test and RTCA (real-time cell analyser, Roche). All three methods proved the ability of uridine and uridine-thymidine combination to protect keratinocytes against 5-FU damagein vitro. While thymidine alone did not show any remarkable effect, the thymidine-uridine combination demonstrated enhanced protective activity compared to uridine alone. Our findings provided the supporting rationale for using uridine or uridine-thymidine ointments in the HF syndrome local therapy.

scholarly journals An integrated systems biology approach to understanding the rules of keratinocyte colony formation

2007 ◽  
Vol 4 (17) ◽  
pp. 1077-1092 ◽  
Author(s):  
Tao Sun ◽  
Phil McMinn ◽  
Simon Coakley ◽  
Mike Holcombe ◽  
Rod Smallwood ◽  
...  
Keyword(s):  
Human Keratinocytes ◽  
Self Organization ◽  
Hacat Cells ◽  
Agent Based ◽  
Wound Model ◽  
Cell Substrate ◽  
Scratch Wound ◽  
Keratinocyte Cell Line Hacat ◽  
Keratinocyte Cell

Closely coupled in vitro and in virtuo models have been used to explore the self-organization of normal human keratinocytes (NHK). Although it can be observed experimentally, we lack the tools to explore many biological rules that govern NHK self-organization. An agent-based computational model was developed, based on rules derived from literature, which predicts the dynamic multicellular morphogenesis of NHK and of a keratinocyte cell line (HaCat cells) under varying extracellular Ca ++ concentrations. The model enables in virtuo exploration of the relative importance of biological rules and was used to test hypotheses in virtuo which were subsequently examined in vitro . Results indicated that cell–cell and cell–substrate adhesions were critically important to NHK self-organization. In contrast, cell cycle length and the number of divisions that transit-amplifying cells could undergo proved non-critical to the final organization. Two further hypotheses, to explain the growth behaviour of HaCat cells, were explored in virtuo —an inability to differentiate and a differing sensitivity to extracellular calcium. In vitro experimentation provided some support for both hypotheses. For NHKs, the prediction was made that the position of stem cells would influence the pattern of cell migration post-wounding. This was then confirmed experimentally using a scratch wound model.

scholarly journals Wound healing promoting activity of Earthworm, Eutyphoeus gammiei (Beddard): in vitro studies on human skin keratinocyte cell line (HaCat).

2018 ◽  
Vol 8 (6) ◽  
pp. 155-158
Author(s):  
Susmita Saha ◽  
Deepjyoti Bhattacharjee ◽  
Anwesha Saha ◽  
Gahin De ◽  
Partha Saha ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Wound Healing ◽  
Cell Line ◽  
Key Factors ◽  
Cell Proliferation And Migration ◽  
Keratinocyte Cell Line Hacat ◽  
Keratinocyte Cell ◽  
And Migration ◽  
Proliferation And Migration

Earthworm, Eutyphoeus gammiei, homogenate (EGH) was screened for wound healing activity on human keratinocyte cell line, HaCat, by cell proliferation and migration assays. The maximum proliferation and migration of keratinocyte cells were observed at the dose of 25μg/ml. As cell proliferation and migration are key factors for wound healing, the study clearly suggests the potential role of earthworm species Eutyphoeus gammiei on wound healing. Keywords: Eutyphoeus gammiei, Keratinocyte, MTT assay, scratch assay.

scholarly journals The Effect of Crude Extracts of Sonchus oleraceus on Cancer Cell Growth (In vitro)

2010 ◽  
Vol 34 (2) ◽  
pp. 30-38
Author(s):  
Zainab R. Zghair
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
In Vitro Study ◽  
Ethanolic Extract ◽  
Cell Types ◽  
Cancer Cell Lines ◽  
Mammary Adenocarcinoma ◽  
Sonchus Oleraceus

This study was designed to evaluate the anticancer, effects of the ethanolic (EE), cold aqueous (CAE), and hot aqueous (HAE) extracts of Sonchus oleraceus on cancer cell lines (in vitro). In vitro study was performed on three cancer cell lines (murine mammary adenocarcinoma AMN-3 cell line, laryngeal carcinoma Hep-2 cell line) and rat embryogenic fibroblast (REF) as normal cell line. Periods of exposure of cell lines were measured at 24, 48, and 72-hr in a microtitration plate under complete sterile conditions. Different concentrations starting from (78.125 to 10000) μg/ml of two fold dilution for each extract were prepared and tested on each cell line, with three replicates for each concentration. The three extracts showed concentration and time dependence with growth inhibitory effects, and the highest effect was obtained from ethanolic extract at higher concentrations after 48 hr. of exposures on both AMN3 and Hep-2 cell lines, while the cytotoxic effect of both cold aqueous and hot aqueous extracts on AMN-3 and Hep-2 cell lines exhibited that the higher concentrations gave a significantly (P<0.05) and the higher inhibition growth rate of cells were increased at 24 hrs.Conclusion: These results suggest that the cytotoxic concentrations of Sonchus oleraceus extracts showed variation in values among cell lines according to cell types in vitro.

scholarly journals Edible flowers — antioxidant activity and impact on cell viability

2013 ◽  
Vol 8 (10) ◽  
pp. 1023-1031 ◽  
Author(s):  
Zdenka Kucekova ◽  
Jiri Mlcek ◽  
Petr Humpolicek ◽  
Otakar Rop
Keyword(s):  
Antioxidant Activity ◽  
Cell Viability ◽  
Cell Line ◽  
Phenolic Compound ◽  
Dry Matter ◽  
Total Phenolic ◽  
Keratinocyte Cell Line Hacat ◽  
Keratinocyte Cell ◽  
Allium Schoenoprasum ◽  
First Time

AbstractThe phenolic compound composition, antioxidant activity and impact on cell viability of edible flower extracts of Allium schoenoprasum; Bellis perennis; Cichorium intybus; Rumex acetosa; Salvia pratensis; Sambucus nigra; Taraxacum officinale; Tragopogon pratensis; Trifolium repens and Viola arvensis was examined for the first time. Total phenolic content of the flowers of these plants fell between 11.72 and 42.74 mg of tannin equivalents/kg of dry matter. Antioxidant activity ranged from 35.56 to 71.62 g of ascorbic acid equivalents/kg of dry matter. Using the Human Hepatocellular Carcinoma cell-line (HepG2) and the Human Immortalized Non-tumorigenic Keratinocyte cell line (HaCaT), we assessed cell viability following a 3 day incubation period in media containing 25, 50, 75 and 100 μg/ml of total phenolic compounds using a colorimetric MTT assay. These three properties could make the herbs useful in treatment of various diseases like cancer. The tested extracts had significant effects on cell viability, but the effects were dependent not only on the phenolic compound concentration and the edible flowers species, but also on the phenolic compound and antioxidant profiles. In addition, responses differed between cell lines.

scholarly journals lncRNA CASC19 Contributes to Radioresistance of Nasopharyngeal Carcinoma by Promoting Autophagy via AMPK-mTOR Pathway

2021 ◽  
Vol 22 (3) ◽  
pp. 1407
Author(s):  
Hongxia Liu ◽  
Wang Zheng ◽  
Qianping Chen ◽  
Yuchuan Zhou ◽  
Yan Pan ◽  
...  
Keyword(s):  
Nasopharyngeal Carcinoma ◽  
Cell Line ◽  
Malignant Tumors ◽  
Underlying Mechanism ◽  
Mtor Pathway ◽  
Rna Seq ◽  
Cellular Autophagy ◽  
First Time

Nasopharyngeal carcinoma (NPC) is one of the most frequent head and neck malignant tumors and is majorly treated by radiotherapy. However, radiation resistance remains a serious obstacle to the successful treatment of NPC. The aim of this study was to discover the underlying mechanism of radioresistance and to elucidate novel genes that may play important roles in the regulation of NPC radiosensitivity. By using RNA-seq analysis of NPC cell line CNE2 and its radioresistant cell line CNE2R, lncRNA CASC19 was screened out as a candidate radioresistance marker. Both in vitro and in vivo data demonstrated that a high expression level of CASC19 was positively correlated with the radioresistance of NPC, and the radiosensitivity of NPC cells was considerably enhanced by knockdown of CASC19. The incidence of autophagy was enhanced in CNE2R in comparison with CNE2 and another NPC cell line HONE1, and silencing autophagy with LC3 siRNA (siLC3) sensitized NPC cells to irradiation. Furthermore, CASC19 siRNA (siCASC19) suppressed cellular autophagy by inhibiting the AMPK/mTOR pathway and promoted apoptosis through the PARP1 pathway. Our results revealed for the first time that lncRNA CASC19 contributed to the radioresistance of NPC by regulating autophagy. In significance, CASC19 might be a potential molecular biomarker and a new therapeutic target in NPC.

scholarly journals Enhancing Anti-Tumoral Potential of CD-NHF by Modulating PI3K/Akt Axis in U87 Ex Vivo Glioma Model

2021 ◽  
Vol 22 (8) ◽  
pp. 3873
Author(s):  
Gabriel Luta ◽  
Mihail Butura ◽  
Adrian Tiron ◽  
Crina E. Tiron
Keyword(s):  
Cell Line ◽  
Ex Vivo ◽  
Glioma Cells ◽  
In Vitro Study ◽  
Immunofluorescent Staining ◽  
Expression Levels ◽  
Disease Diagnostics ◽  
Phosphorylated Akt ◽  
Significant Impairment

Background: In the latest years, there has been an increased interest in nanomaterials that may provide promising novel approaches to disease diagnostics and therapeutics. Our previous results demonstrated that Carbon-dots prepared from N-hydroxyphthalimide (CD-NHF) exhibited anti-tumoral activity on several cancer cell lines such as MDA-MB-231, A375, A549, and RPMI8226, while U87 glioma tumor cells were unaffected. Gliomas represent one of the most common types of human primary brain tumors and are responsible for the majority of deaths. In the present in vitro study, we expand our previous investigation on CD-NHF in the U87 cell line by adding different drug combinations. Methods: Cell viability, migration, invasion, and immunofluorescent staining of key molecular pathways have been assessed after various treatments with CD-NHF and/or K252A and AKTVIII inhibitors in the U87 cell line. Results: Association of an inhibitor strongly potentiates the anti-tumoral properties of CD-NHF identified by significant impairment of migration, invasion, and expression levels of phosphorylated Akt, p70S6Kinase, or by decreasing expression levels of Bcl-2, IL-6, STAT3, and Slug. Conclusions: Using simultaneously reduced doses of both CD-NHF and an inhibitor in order to reduce side effects, the viability and invasiveness of U87 glioma cells were significantly impaired.

scholarly journals Analysis of a Preliminary microRNA Expression Signature in a Human Telangiectatic Osteogenic Sarcoma Cancer Cell Line

2021 ◽  
Vol 22 (3) ◽  
pp. 1163
Author(s):  
Gaia Palmini ◽  
Cecilia Romagnoli ◽  
Simone Donati ◽  
Roberto Zonefrati ◽  
Gianna Galli ◽  
...  
Keyword(s):  
Cell Line ◽  
Molecular Mechanisms ◽  
Osteogenic Sarcoma ◽  
Cancer Cell Line ◽  
Microscopic Features ◽  
In Vitro Establishment ◽  
Profile Characteristic ◽  
First Time

Telangiectatic osteosarcoma (TOS) is an aggressive variant of osteosarcoma (OS) with distinctive radiographic, gross, microscopic features, and prognostic implications. Despite several studies on OS, we are still far from understanding the molecular mechanisms of TOS. In recent years, many studies have demonstrated not only that microRNAs (miRNAs) are involved in OS tumorigenesis, development, and metastasis, but also that the presence in high-grade types of OS of cancer stem cells (CSCs) plays an important role in tumor progression. Despite these findings, nothing has been described previously about the expression of miRNAs and the presence of CSCs in human TOS. Therefore, we have isolated/characterized a putative CSC cell line from human TOS (TOS-CSCs) and evaluated the expression levels of several miRNAs in TOS-CSCs using real-time quantitative assays. We show, for the first time, the existence of CSCs in human TOS, highlighting the in vitro establishment of this unique stabilized cell line and an identification of a preliminary expression of the miRNA profile, characteristic of TOS-CSCs. These findings represent an important step in the study of the biology of one of the most aggressive variants of OS and the role of miRNAs in TOS-CSC behavior.

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