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Shun Sawatsubashi, Yudai Joko, Seiji Fukumoto, Toshio Matsumoto, Shigeo S Sugano
CRISPR/Cas9-based genome editing has dramatically accelerated genome engineering. An important aspect of genome engineering is efficient knock-in technology. For improved knock-in efficiency, the non-homologous end joining (NHEJ) repair pathway has been used over the homology-dependent repair pathway, but there remains a need to reduce the complexity of the preparation of donor vectors. We developed the versatile NHEJ-based knock-in module for genome editing (VIKING). Using the consensus sequence of the time-honored pUC vector to cut donor vectors, any vector with a pUC backbone could be used as the donor vector without customization...
January 12, 2018: Scientific Reports
Mitsutoki Hatta, Yuki Miyake, Kunitoshi Uchida, Jun Yamazaki
Epithelial-mesenchymal transition (EMT) is a biological event in which epithelial cells lose their polarity and cell-cell adhesions and concomitantly acquire mesenchymal traits, and is thought to play an important role in pathological processes such as wound healing and cancer progression. In this study, we evaluated transforming growth factor (TGF)-β1-treated human keratinocyte HaCaT cells as an in vitro model of EMT. HaCaT cells were changed into an elongated fibroblast-like morphology, which is indicative of EMT in response to TGF-β1...
January 8, 2018: Biochemical and Biophysical Research Communications
Laila Al-Eryani, Sabine Waigel, Ashish Tyagi, Jana Peremarti, Samantha F Jenkins, Chendil Damodaran, J Christopher States
Background: Arsenic is a widely distributed toxic natural element. Chronic arsenic ingestion causes several cancers, especially skin cancer. Arsenic-induced cancer mechanisms are not well defined, but several studies indicate that mutation is not the driving force and that microRNA expression changes play a role. Chronic low arsenite exposure malignantly transforms immortalized human keratinocytes (HaCaT), serving as a model for arsenic-induced skin carcinogenesis. Hypothesis: Early changes in miRNA expression in HaCaT cells chronically exposed to arsenite will reveal early steps in transformation...
January 8, 2018: Toxicological Sciences: An Official Journal of the Society of Toxicology
Dohyun Lee, Jinsun Lim, Kyung-Chul Woo, Kyong-Tai Kim
Epidermal growth factor (EGF) stimulates cell growth, proliferation, and survival. The biological benefits of EGF have been utilized in medical uses for improving wound healing as well as in today's skin cosmetics. EGF has been found in urine, saliva, milk, and plasma, but its efficient isolation remains a difficult task. With technical advances, recombinant protein purification technique has been used for EGF production. However, the recombinant EGF is still expensive and keeping it with stable activity is difficult to be used widely...
January 9, 2018: Scientific Reports
Eunji Kim, Kyeonghwan Hwang, Jongsung Lee, Sang Yun Han, Eun-Mi Kim, Junseong Park, Jae Youl Cho
Epigallocatechin gallate (EGCG) is a catechin and an abundant polyphenol in green tea. Although several papers have evaluated EGCG as a cosmetic constituent, the skin hydration effect of EGCG is poorly understood. We aimed to investigate the mechanism by which EGCG promotes skin hydration by measuring hyaluronic acid synthase (HAS) and hyaluronidase (HYAL) gene expression and antioxidant and anti-pigmentation properties using cell proliferation assay, Western blotting analysis, luciferase assay, 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, and reverse transcription polymerase chain reaction (RT-PCR) analysis...
January 6, 2018: International Journal of Molecular Sciences
Sonia Iranpour, Nasser Mahdavi-Shahri, Raheleh Miri, Halimeh Hasanzadeh, Hamid Reza Bidkhori, Hojjat Naderi-Meshkin, Ehsan Zahabi, Maryam M Matin
Human amniotic membrane (HAM) has been widely used as a natural scaffold in tissue engineering due to many of its unique biological properties such as providing growth factors, cytokines and tissue inhibitors of metalloproteinases. This study aimed at finding the most suitable and supportive layer of HAM as a delivery system for autologous or allogeneic cell transplantation. Three different layers of HAM were examined including basement membrane, epithelial and stromal layers. In order to prepare the basement membrane, de-epithelialization was performed using 0...
January 8, 2018: Cell and Tissue Banking
Karol Sikora, Maciej Jaśkiewicz, Damian Neubauer, Marta Bauer, Sylwia Bartoszewska, Wioletta Barańska-Rybak, Wojciech Kamysz
In view of an appreciable increase in resistance of Staphylococcus aureus to the conventional antibiotics, it is desired to develop new effective drugs. Antimicrobial peptides (AMPs) seem to be attractive candidates. In general, AMPs samples used for in vitro studies consist of a peptide, counter-ion, and water. The presence of the counter-ion could be significant as it affects peptide secondary structure and biological activity. The purpose of this study was to estimate the impact of counter-ion on antistaphylococcal activity of selected AMPs (CAMEL, citropin 1...
January 6, 2018: Amino Acids
Catiúscia P de Oliveira, Sabrina L Büttenbender, Willian A Prado, Aline Beckenkamp, Ana C Asbahr, Andréia Buffon, Silvia S Guterres, Adriana R Pohlmann
Methotrexate is a folic acid antagonist and its incorporation into nanoformulations is a promising strategy to increase the drug antiproliferative effect on human breast cancer cells by overexpressing folate receptors. To evaluate the efficiency and selectivity of nanoformulations containing methotrexate and its diethyl ester derivative, using two mechanisms of drug incorporation (encapsulation and surface functionalization) in the in vitro cellular uptake and antiproliferative activity in non-tumoral immortalized human keratinocytes (HaCaT) and in human breast carcinoma cells (MCF-7)...
January 4, 2018: Nanomaterials
Jian Zhang, Na Li, Kun Zhang, Ping Pan, Hui Zhang, Ning Li, Qiong Tian, Li Zhang, Wenli Liu, Tianyuan Gao, Xin Mu, Dengchao Feng, Wei Guo, Xiaona Ma
Actinic keratosis (AK), is the most common precancerous skin lesion induced by the excessive ultraviolet B (UVB) and is a significant threat to the public health. UVB exposure causes oxidative DNA damage and is considered to be a significant contributor to AK and subsequent development of skin cancer. Besides, activation of p38 MAPK also plays a significant role in the development of AK. In our clinical findings revealed that p38 MAPK, phospho-MSK1, and γ-H2AX were significantly highly expressed in human AK tissue than the normal healthy skin tissue...
December 29, 2017: Anti-cancer Agents in Medicinal Chemistry
Kuen-Tze Lin, Tsu-Chung Chang, Feng-Yi Lai, Chun-Shu Lin, Hsing-Lung Chao, Shih-Yu Lee
Skin injury is a major complication during radiation therapy and is associated with oxidative damage to skin cells. An effective and safe radioprotectant to prevent this skin damage is still unavailable. The Rhodiola crenulata root extract (RCE) has been reported to be a free radical scavenger and a potent anti-oxidant in both in vitro and in vivo models. In the current study, we investigated the effects of RCE on ionizing radiation-induced skin injury and its underlying mechanisms. HaCaT cells - a non-cancerous skin cell line together with HepG2, Caco2, A549, and OECM cancer cell lines - were pre-treated with RCE for 24[Formula: see text]h followed by exposure to 15 Gy using Caesium-137 as a γ-ray source...
January 3, 2018: American Journal of Chinese Medicine
Patricia Milagros Maquera Huacho, Marianne N Marques Nogueira, Fernanda G Basso, Miguel Jafelicci Junior, Renata S Francisconi, Denise M P Spolidorio
The aim of this study was to evaluate the surface free energy (SFE), wetting and surface properties as well as antimicrobial, adhesion and biocompatibility properties of diamond-like carbon (DLC)-coated surfaces. In addition, the leakage of Escherichia coli through the abutment-dental implant interface was also calculated. SFE was calculated from contact angle values; R a was measured before and after DLC coating. Antimicrobial and adhesion properties against E. coli and cytotoxicity of DLC with human keratinocytes (HaCaT) were evaluated...
May 2017: Brazilian Dental Journal
Yoon Jeong Nam, Chung Soo Lee
Protocatechuic acid has demonstrated to have antioxidant and anti-inflammatory effects. We assessed whether protocatechuic acid may reduce the inflammatory mediator production, which is regulated by the Toll-like receptor-4-dependent Akt, mTOR and NF-κB pathway, and JNK and p38-MAPK in HaCaT cells and primary keratinocytes. Protocatechuic acid, Akt inhibitor, Bay 11-7085 and N-acetylcysteine reduced the lipopolysaccharide-caused production of cytokines and chemokines, expression of cyclooxygenase, increase in the levels and activities of Toll-like receptor-4, p-Akt and mTOR, activation of NF-κB, phosphorylation of the JNK and p38-MAPK, and production of reactive oxygen species in keratinocytes...
December 29, 2017: International Immunopharmacology
Tomasz Stefański, Renata Mikstacka, Rafał Kurczab, Zbigniew Dutkiewicz, Małgorzata Kucińska, Marek Murias, Małgorzata Zielińska-Przyjemska, Michał Cichocki, Anna Teubert, Mariusz Kaczmarek, Adam Hogendorf, Stanisław Sobiak
A series of novel combretastatin A-4 (CA-4) thio derivatives containing different molecular cores, namely α-phenylcinnamic acids (core 1), (Z)-stilbenes (core 2), 4,5-disubstituted oxazoles (core 3), and 4,5-disubstituted N-methylimidazoles (core 4), as cis-restricted analogues were designed and synthesized. They were selected with the use of a parallel virtual screening protocol including the generation of a virtual combinatorial library based on an elaborated synthesis protocol of CA-4 analogues. The selected compounds were evaluated for antiproliferative activity against a panel of six human cancer cell lines (A431, HeLa, MCF7, MDA-MB-231, A549 and SKOV) and two human non-cancer cell lines (HaCaT and CCD39Lu)...
December 12, 2017: European Journal of Medicinal Chemistry
Dalia H Abdelkader, Mohamed A Osman, Sanaa A El-Gizawy, Suzan J Hawthorne, Ahmed M Faheem, Paul A McCarron
We describe the development of a nanoparticulate system, with variation of poly(ethylene glycol) (PEG) content, capable of releasing therapeutic levels of bioactive insulin for extended periods of time. Recombinant human insulin was encapsulated in poly(d,l-lactide-co-glycolide) nanoparticles, manufactured with variation in poly(ethylene glycol) content, and shown to be stable for 6days using SDS-PAGE, western blot and MALDI MS. To determine if insulin released from this sustained release matrix could stimulate migration of cell types normally active in dermal repair, a model wound was simulated by scratching confluent cultures of human keratinocytes (HaCaT) and fibroblasts (Hs27)...
December 26, 2017: European Journal of Pharmaceutical Sciences
Xin Zhang, Xia Liu, Shuxia Kang, Caiyun Liu, Yuqin Hao
AIMS AND BACKGROUND: Squamous cell carcinoma (SCC) is one of the most common skin cancers. Photodynamic therapy (PDT) is a non-invasive treatment for SCC, but it is usually effective only on tumors just under the skin. Resveratrol (Res) is a polyphenolic compound, which is capable of promoting apoptosis of a variety of cancer cells. Res administration is non-invasive and effective on SCC, thus it may be used as an adjuvant for PDT. So far, there is no published study investigating the combination use of PDT with Res to improve clinical outcome of SCC...
December 23, 2017: Cancer Biomarkers: Section A of Disease Markers
Wenming Wang, Xiaoling Yu, Chao Wu, Hongzhong Jin
The predominant role of Wnt family member 5A (Wnt5a) is to induce non-canonical Wnt signalling pathways, including the Wnt‑Ca2+ and Wnt‑planar cell polarity pathways. Enhanced Wnt5a expression is involved in the formation of psoriatic plaques; however, its mechanistic role remains to be determined. In the present study, the effects of Wnt5a expression on HaCaT keratinocytes were investigated. HaCaT cells were cultured in medium supplemented with 0, 40 or 80 ng/ml Wnt5a for 24 h. Cell proliferation, the cell cycle, gene expression and inflammatory responses were investigated using Cell‑Counting Kit‑8 assays, flow cytometry analyses, reverse transcription‑quantitative polymerase chain reaction analyses and enzyme‑linked immunosorbent assays, respectively...
December 27, 2017: Molecular Medicine Reports
Jinghong Yu, Zhicai Xiao, Ruizhi Zhao, Chuanjian Lu, Yuemei Zhang
Astilbin has anti-inflammatory and immunoregulatory effects, and is frequently used in prescriptions treating psoriasis; however, the mechanism remains to be fully elucidated. In the present study, the effect of an astilbin microemulsion on a psoriasis‑like model in guinea pigs was examined, and the underlying mechanism was investigated. The levels of interkeukin (IL)‑6, IL‑17A and IL‑22 were determined using fluorescent reverse transcription‑quantitative polymerase chain reaction analysis and enzyme‑linked immunosorbent assays...
December 22, 2017: Molecular Medicine Reports
Yue Deng, Weiyuan Ma
Substantial preclinical evidence has indicated out a direct anti‑proliferation effect of metformin on various solid tumors; however, further and more detailed exploration into its molecular mechanism remains to be performed. The present study aimed to investigate the effect of metformin on cell viability and its underlying mechanism, in the cultured human skin keratinocyte cell line, HaCaT. In addition, it aimed to clarify the role of the microRNA-21(miR-21)/phosphatase and tensin homolog (PTEN)/AKT serine/threonine kinase 1 (Akt) signaling pathway, which has been hypothesized to be involved in the molecular mechanism of this drug...
December 27, 2017: Molecular Medicine Reports
Jangho Lee, Su Jeong Ha, Joon Park, Yong Ho Kim, Nam Hyouck Lee, Young Eon Kim, Yoonsook Kim, Kyung-Mo Song, Sung Keun Jung
1,8-cineole is a natural monoterpene cyclic ether present in Eucalyptus, and has been reported to exhibit anti-inflammatory and antioxidant effects. However, the preventive effect of 1,8-cineole on skin carcinogenesis and the molecular mechanism of action responsible remains unknown. In the present study, we investigated the effect of 1,8-cineole on UVB-induced skin carcinogenesis. 1,8-cineole inhibited UVB-induced cyclooxygenase-2 (COX-2) protein and mRNA expression and prostaglandin E2 (PGE2) generation in HaCaT cells...
December 1, 2017: Oncotarget
Rui Wang, Zigang Zhao, Liqiang Zheng, Xiaojing Xing, Wei Ba, Junfen Zhang, Min Huang, Wenwei Zhu, Bing Liu, Xianfu Meng, Jia Bai, Chengxin Li, Hengjin Li
Psoriasis is a chronic inflammatory disease of the skin for which an effective treatment strategy remains to be developed. Characteristics of psoriasis include an altered differentiation of keratinocytes and hyperplasia of the skin. The present study aimed to investigate the role served by miR-520a in psoriasis. The results demonstrated that miR-520a inhibited the proliferation of HaCaT cells. miR-520a directly regulated the mRNA and protein expression of its target gene, protein kinase B (AKT). The siRNA silencing of AKT expression in these cells was also evaluated...
December 2017: Experimental and Therapeutic Medicine
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