• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.35 no.4
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• pp.199-204
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• 2009

Purpose: The purpose of this study is to investigate the epithelial-mesenchymal transition (EMT) induced by Snail transcription factor and Snail-transfected in vivo tumors with histopathological features. Materials and methods: We induced in vivo xenografted tumorigenesis in the oral vestibules of nude mice by a Snail transfected HaCaT cell line and investigated morphological and immunohistochemical features in Snail expressive tumors. Results: We identified tumor masses in 14 out of 15 nude mice in the HaCaT-Snail cell inoculation group, but no tumors were present in any of the HaCaT cell inoculation group. Induced tumors showed features of poorly differentiated carcinoma with invasion to neighboring muscles and bones. The HaCaT-Snail tumors showed decreased expressions of E-cadherin and cytokeratin, but showed increased expressions of vimentin and N-cadherin. Discussion: The Snail transfected xenograft can improve productivity of malignant tumors, show various histopathological features including invasive growth, and aid in the investigation of tumor progression and the interaction with surrounding tissues.

• Journal of Korean society of Dental Hygiene
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• v.15 no.4
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• pp.719-725
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• 2015

Objectives: The purpose of this study was to determine the toxic effects of sodium lauryl sulfate(SLS) in human keratinocyte HaCaT cells and mouse fibroblast NIH-3T3 cells. Methods: The effect of sodium lauryl sulfate(SLS) cell viability and proliferation were determined by WST-1 assay and changes shape of nucleus were evaluated by Hoechst staining under fluorescence microscopy. Additionally, observation of cell morphological changes under light microscopy. Results: SLS induced cytotoxicity and a marked apoptosis in both HaCaT and NIH-3T3 cell lines. With the result of the WST-1 assay, SLS induced the cytotoxicity of 0.005% and 0.0075%, 0.01% SLS for 24 h after HaCaT and NIH-3T3 cells in time and dose-dependent manner(p<0.005). SLS inhibited cell growth and caused apoptosis as evidenced by nuclear fragmentation and condensation. Thus, determination of the morphological changes to define apoptosis was visualized using inverted phase contrast microscopy. Conclusions: SLS had toxicity of the human keratinocyte cells and mouse fibroblast cells and this study will provide the basic data for the development of proper SLS concentration in dentifrice.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.15 no.11
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• pp.6774-6781
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• 2014

This study examined the efficacy and the mechanism of action of biological response modifiers, ginsenosides Rb1 and Rg1 isolated from Panax ginseng C.A. Meyer on human keratinocytes HaCaT cell lines. A non-significant cytotoxic response was obtained in the HaCaT cell lines on treatment with various concentrations of ginsenosides Rb1 and Rg1 for different time durations. Furthermore, the global changes in the mRNA profile of HaCaT cells were investigated using DNA microarrays after stimulation with the ginsenosides Rb1 and Rg1. Ginsenosides Rb1 and Rg1 strongly increased FGF2 in HaCaT cells, and were found to be a candidate gene for antioxidant activity and elasticity. Other key candidate genes for antioxidant activity, such as FANCD2, LEPR, and FAS, also show enhanced regulation in HaCaT cells treated with ginsenoside Rb1. This study will be useful for understanding the regulatory genes involved in skin elasticity and signal transduction pathway stimulated by the ginsenoside Rb1. This paper currently focuses on the key factors regulating the interaction of anti-aging principles and skin elasticity.

• Journal of Haehwa Medicine
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• v.28 no.2
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• pp.41-47
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• 2019

Objectives : Atopic dermatitis is an irritable skin disease accompanying rash and itching leading to impaired skin barrier. ATO-ALL is an ethanol extract of natural products comprising 12 herbs and effective on atopic dermatitis. In this study, we aimed to propose that the effect of ATO-ALL on skin regeneration in human keratinocyte cell line, HaCaT cells. Methods : To evaluate the skin regenerating effects of ATO-ALL, scratch wound healing assay, bromodeoxyuridine (BrdU) assay, and propidum iodide (PI) assay were performed using cultured HaCaT cell line. Result : Scratch wound healing assay showed that ATO-ALL was able to enhance the gap filling activity more than 2-fold at 7 ppm concentration compared with control group. BrdU assay demonstrated that ATO-ALL treatment increased the de novo cell proliferation in a dose-dependent manner. Finally, PI assay indicated that the cell cycle of HaCaT cells was modulated by ATO-ALL treatment. Conclusions : These results suggested that ATO-ALL may have skin regenerating effects by increasing cell proliferation via cell cycle regulation. Taken together, ATO-ALL is supposed to have a potential on regeneration of damaged skin or functional disease including atopic dermatitis.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.5
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• pp.2015-2020
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• 2012

Oleanolic acid (OA) is a naturally occurring triterpenoid in food materials and is a component of the leaves and roots of Olea europaea, Viscum album L., Aralia chinensis L. and more than 120 other plant species. There are several reports validating its antitumor activity against different cancer cells apart from its hepatoprotective activity. However, antitumor activity against skin cancer has not beed studied well thus far. Hence the present study of effects of OA against HaCaT (immortalized keratinocyte) cells - a cell-based epithelial model system for toxicity/ethnopharmacology-based studies - was conducted. Radical scavenging activity ($DPPH{\cdot}$) and FRAP were determined spectrophotometrically. Proliferation was assessed by XTT assay at 24, 48 and 72 hrs with exposure to various concentrations (12.5-200 ${\mu}M$) of OA. Apoptotic induction potential of OA was demonstrated using a cellular DNA fragmentation ELISA method. Morphological studies were also carried out to elucidate its antitumor potential. The results revealed that OA induces apoptosis by altering cellular morphology as well as DNA integrity in HaCaT cells in a dose-dependent manner, with comparatively low cytotoxicity. The moderate toxicity observed in HaCaT cells, with induction of apoptosis, possibly suggests greater involvement of programmed-cell death-mediated mechanisms. We conclude that OA has relatively low toxicity and has the potential to induce apoptosis in HaCaT cells and hence provides a substantial and sound scientific basis for further validation studies.

• The Journal of Pediatrics of Korean Medicine
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• v.30 no.2
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• pp.96-106
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• 2016

Objectives The purpose of this study is to investigate the effects of Gleditsiae Fructus 70% EtOH extract (JS_E) on anti-inflammatory action in HaCaT cells (A spontaneously immortalized human keratinocyte cell line) and inhibiting triglyceride genesis in SEB-1 cells (Immortalized human sebocyte). Methods The anti-inflammatory effect of JS_E was analyzed by enzyme-linked immunosorbent assays (ELISA) which measured levels of IP-10, RANTES and MDC in HaCaT cells. Also the effect on secretion of sebum of JS_E was analyzed by TG-S kit which measured the quantity of triglyceride in SEB-1 cells. Results JS_E inhibited IP-10, RANTES and MDC expression in a dose dependent manner. IP-10 expression was inhibited significantly in comparison to TNF-${\alpha}$ and IFN-${\gamma}$ recombination (TI) control group at concentration of JS_E $200{\mu}g/ml$ and RANTES and MDC expressions were inhibited significantly at concentration of JS_E 100, $200{\mu}g/ml$. JS_E also inhibited triglyceride secretion of SEB-1 cells significantly in comparison to the control group in a dose dependent manner. Conclusions This study shows that JS_E has the effects of anti-inflammatory action and inhibiting sebum secretion. According to these results, JS_E can be used for treating skin diseases such as acne and dermatitis caused by inflammation and excessive secretion of sebum by controlling the activity of the HaCaT and SEB-1 cells.

• BMB Reports
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• v.48 no.9
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• pp.495-500
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• 2015

Up-regulation of cell adhesion molecules and proinflammatory cytokines contributes to enhanced monocyte adhesiveness and infiltration into the skin, during the pathogenesis of various inflammatory skin diseases, including atopic dermatitis. In this study, we examined the anti-inflammatory effects of butein, a tetrahydroxychalcone, and its action mechanisms using TNF-α-stimulated keratinocytes. Butein significantly inhibited TNF-α-induced ICAM-I expression and monocyte adhesion in human keratinocyte cell line HaCaT. Butein also decreased TNF-α-induced pro-inflammatory mediators, such as IL-6, IP-10 and MCP-1, in HaCaT cells. Butein decreased TNF-α-induced ROS generation in a dose-dependent manner in HaCaT cells. In addition, treatment of HaCaT cells with butein suppressed TNF-α-induced MAPK activation. Furthermore, butein suppressed TNF-α-induced NF-kappaB activation. Overall, our results indicate that butein has immunomodulatory activities by inhibiting expression of proinflammatory mediators in keratinocytes. Therefore, butein may be used as a therapeutic agent for the treatment of inflammatory skin diseases. [BMB Reports 2015; 48(9): 495-500]

• Journal of Acupuncture Research
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• v.34 no.1
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• pp.23-30
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• 2017

Objectives : In this study, the roles of Interleukin (IL)-4 and Signal transducer and activator of transcription 6 (STAT6), which have been reported to play a role in the pathogenesis of inflammation and cancer, were evaluated in snake venom toxin (SVT)-induced apoptosis. Methods : Inflammation was induced in human HaCaT kerationocytes, by lipopolysaccharide (LPS; $1{\mu}g/mL$) or tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), followed by treatment with SVT (0, 1, or $2{\mu}g/mL$). Cell viability was assessed by MTT assays after 24 h, and the expression of levels of IL-4, STAT6, and the apoptosis-related proteins p53, Bax, and Bcl-2 were evaluated by western blotting. Electro mobility shift assays (EMSAs) were performed to evaluate the DNA binding capacity of STAT6. Results : MTT assays showed that inflammation-induced growth of HaCaT cells following LPS or TNF-${\alpha}$ stimulation was inhibited by SVT. Western blot analysis showed that p53 and Bax, which promote apoptosis, were increased, whereas that of Bcl-2, an anti-apoptotic protein, was decreased in a concentration-dependent manner in LPS- or TNF-${\alpha}$-induced HaCaT cells following treatment with SVT. Moreover, following treatment of HaCaT cells with LPS, IL-4 concentrations were increased, and treatment with SVT further increased IL-4 expression in a concentration-dependent manner. Western blotting and EMSAs showed that the phosphorylated form of STAT6 was increased in HaCaT cells in the context of LPS- or TNF-${\alpha}$-induced inflammation in a concentration-dependent manner, concomitant with an increase in the DNA binding activity of STAT6. Conclusion : SVT can effectively promote apoptosis in HaCaT cells in the presence of inflammation through a pathway involving IL-4 and STAT6.

• The Journal of Korean Medicine
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• v.43 no.3
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• pp.1-15
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• 2022

Objectives: Myrrh have been used as a traditional remedy to treat infectious and inflammatory diseases. However, it is largely unknown whether myrrh ethanol extract could exhibit the inhibitory activities against particulate matter (PM)-induced skin injury on human keratinocytes, HaCaT cells. Therefore, this study was aimed to investigate the inhibitory activity of myrrh ethanol extract on PM-induced skin injury in HaCaT cells. Methods: To investigate the inhibitory effects of myrrh ethanol extract in HaCaT cells, the skin injury model of HaCaT cells was established under PM treatment. HaCaT keratinocyte cells were pre-treated with myrrh ethanol extract for 1 h, and then stimulated with PM. Then, the cells were harvested to measure the cell viability, reactive oxygen species (ROS), pro-inflammatory cytokines including interleukin (IL) 1-beta, IL-6, and tumor necrosis factor (TNF)-𝛼, hyaluronidase, collagen, MMPs. In addition, we examined the mitogen activated protein kinases (MAPKs) and inhibitory kappa B alpha (I𝜅-B𝛼) as inhibitory mechanisms of myrrh ethanol extract. Results: The treatment of myrrh ethanol extract inhibited the PM-induced cell death and ROS production in HaCaT cells. In addition, myrrh ethanol extract treatment inhibited the PM-induced elevation of IL-1beta, IL-6, and TNF-𝛼. Also, myrrh ethanol extract treatment inhibited the increase of hyaluronidase, MMP and decrease of collagen. Furthermore, myrrh ethanol extract treatment inhibited the activation of MAPKs and the degradation of I𝜅-B𝛼. Conclusions: Our result suggest that treatment of myrrh ethanol extract could inhibit the PM-induced skin injury via deactivation of MAPKs and nuclear factor (NF)-𝜅B in HaCaT cells. This study could suggest that myrrh ethanol extract could be a beneficial agent to prevent skin damage or inflammation.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.2
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• pp.631-637
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• 2013

Luteolin is a naturally occurring flavonoid present in many plants with diverse applications in pharmacology. Despite several studies elucidating its significant anti-cancer activity against various cancer cells, the mechanism of action in skin cancer is not well addressed. Hence, we investigated the effects of luteolin in HaCaT (human immortalized keratinocytes) and A375 (human melanoma) cells. The radical scavenging abilities of luteolin were determined spectrophotometrically, prior to a cytotoxic study (XTT assay). Inhibitory effects were assessed by colony formation assay. Further, the capability of luteolin to induce cell cycle arrest and apoptosis were demonstrated by flow cytometry and cellular DNA fragmentation ELISA, respectively. The results revealed that luteolin possesses considerable cytotoxicity against both HaCaT and A375 cells with $IC_{50}$ values of 37.1 ${\mu}M$ and 115.1 ${\mu}M$, respectively. Luteolin also inhibited colony formation and induced apoptosis in a dose and time-dependent manner by disturbing cellular integrity as evident from morphological evaluation by Wright-Giemsa staining. Accumulation of cells in G2/M (0.83-8.14%) phase for HaCaT cells and G0/G1 (60.4-72.6%) phase for A375 cells after 24 h treatment indicated cell cycle arresting potential of this flavonoid. These data suggest that luteolin inhibits cell proliferation and promotes cell cycle arrest and apoptosis in skin cancer cells with possible involvement of programmed cell death, providing a substantial basis for it to be developed into a potent chemopreventive template for skin cancer.