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Guanghong Fu, Dan Gong, Jia Wan
Objective To investigate the effect of E5 protein of human papillomavirus type 16 (HPV16 E5) on keratinocyte growth factor-triggered autophagy. Methods Firstly, pCI-neo-E5 recombinant plasmid was constructed. After that, the recombinant plasmid harboring pCI-neo-E5 was transfected into HaCaT cells using Lipofectamine(TM)2000 reagent, and the mRNA transcriptional level of 16 E5 was evaluated by quantitative real-time PCR (qRT-PCR). Then, the expression of LC3BIIwas detected by Western blotting and the localization of LC3B was observed by immunofluorescence in pCI-neo-E5-HA-transfected HaCaT cells triggered with keratinocyte growth factor...
November 2016: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
Waleska K Martins, Andreza B Gomide, Érico T Costa, Helena C Junqueira, Beatriz S Stolf, Rosangela Itri, Maurício S Baptista
BACKGROUND: Cell senescence is a process of central importance to the understanding of aging as well as to the development of new drugs. It is related with genomic instability, which has been shown to occur in the presence of autophagy deficiency. Yet, the mechanism that triggers genomic instability and senescence from a condition of autophagy deficiency remains unknown. By analyzing the consequences of treating human keratinocytes (HaCaT) with the pentacyclic triterpenoid Betulinic Acid (BA) we were able to propose that cell senescence can develop as a response to parallel damage in the membranes of mitochondria and lysosome...
October 20, 2016: Biochimica et Biophysica Acta
Gabor Nagy, Viktoria Baksa, Alexandra Kiss, Melinda Turani, Gaspar Banfalvi
The toxicity of gadolinium is reduced by chelating agents that render this heavy metal into contrast complexes used for medical magnetic resonance imaging. However, the dissociation of gadolinium chelates is known to generate Gd(3+) ions, the cellular toxicity of which has not been tested in details. The cytotoxic effects of Gd(III) ions were evaluated by monitoring the proliferation, measuring the cellular motility and following chromatin changes in various cell lines upon Gd(3+) treatment. Measurements applied long-term scanning microscopy and a perfusion platform that replaced the medium with test solutions, bypassed physical contact with the cell culture during experiments, and provided uninterrupted high time-resolution time-lapse photomicrography for an extended period of time...
October 22, 2016: Apoptosis: An International Journal on Programmed Cell Death
Jennifer Hennen, Brunhilde Blömeke
In vitro approaches address key steps of chemical-induced skin sensitization but there is uncertainty how keratinocytes, which play a crucial role not only regarding xenobiotic metabolism but also skin inflammation, impact on chemicals' potential and potency of dendritic cell activation. We investigated these aspects by coculturing THP-1 cells, as surrogate dendritic cells, with HaCaT keratinocytes. We tested our HaCaT/THP-1 model with a set of 14 sensitizers, containing 7 prohaptens, and 10 non-sensitizers...
October 21, 2016: ALTEX
Dorimar Stiz, Adriana Campos, Ana Lúcia Tasca Gois Ruiz, João Ernesto de Carvalho, Rogério Corrêa, Valdir Cechinel-Filho
This work describes the antiproliferative potential of 14 cyclic imides (methylphtalimides, carboxylic acid phtalimides and itaconimides) against several human cancer cell lines. The antiproliferative effect was evaluated using the sulforhodamine B assay. Although some compounds from methylphtalimide and carboxylic acid phtalimide classes exhibited a selective antiproliferative activity, the itaconimides (11-14) exhibited the best results, especially compound 14, which presented a TGI (concentration that produces total growth inhibition) value of 0...
October 21, 2016: Zeitschrift Für Naturforschung. C, A Journal of Biosciences
Ulvi K Gursoy, Mervi Gursoy, Eija Könönen, Herman O Sintim, Veli-Jukka Uitto, Stina Syrjänen
In construction of epithelial cells as multilayers, the cells are grown submerged to confluence on fibroblast-embedded collagen gels and, then, lifted to air to promote their stratification. We recently demonstrated that gingival epithelial cells form uniform monolayers on semi-permeable nitrocellulose membranes, supported with a semi-solid growth medium, which allows the cells to grow at an air-liquid-solid interface from the beginning of the culturing protocol. In this study, the aim was to further develop our previous model to form a multilayered gingival epithelial culture model...
October 17, 2016: Cytotechnology
Tianhui Wu, Xin Chen, Rui Peng, Handeng Liu, Pin Yin, Huimin Peng, Yujian Zhou, Yan Sun, Li Wen, Hong Yi, Ailing Li, Zheng Zhang
Cervical cancer is the second most commonly diagnosed type of cancer among women after breast cancer. Recent research has addressed the role of microRNAs in cervical cancer. In the present study, we aimed to determine the effect of let‑7a on the regulation of the cell proliferation of cervical cancer and the related signaling pathway. Real‑time RT‑PCR was used to detect the expression of let‑7a in the blood of cervical cancer patients and normal controls. The expression of let‑7a was also assessed in cervical cancer cell lines: HeLa, SiHa and normal human immortalized keratinocyes HaCaT...
October 11, 2016: Oncology Reports
Zheng Jun Li, Dae-Kyoung Choi, Kyung-Cheol Sohn, Seul Ki Lim, Myung Im, Young Lee, Young-Joon Seo, Chang Deok Kim, Jeung-Hoon Lee
BACKGROUND: Interleukin-17A (IL-17A) is mainly secreted from Th17 cells that are activated by various stimuli including CpG oligodeoxynucleotide, a Toll-like receptor 9 (TLR9) ligand. Recently, it has been demonstrated that keratinocytes play an important role in the pathogenesis of psoriasis. OBJECTIVE: To investigate the potential role of keratinocytes, we examined whether TLR9 ligand CpG induces IL-17A expression in keratinocytes. METHODS: We used HaCaT keratinocytes as a model system, and determined CpG-induced IL-17A using enzyme-linked immunosorbent assay and Western blot...
October 2016: Annals of Dermatology
Dimple Chouhan, Bijayshree Chakraborty, Samit K Nandi, Biman B Mandal
Bombyx mori silk fibroin (BMSF) as biopolymer has been extensively explored in wound healing applications. However, limited study is available on the potential of silk fibroin (SF) from non-mulberry (Antheraea assama and Philosamia ricini) silk variety. Herein, we have developed non-mulberry SF (NMSF) based electrospun mats functionalized with epidermal growth factor (EGF) and ciprofloxacin HCl as potential wound dressing. The NMSF based mats exhibited essential properties of wound dressing like biocompatibility, high water retention capacity (440%), water vapour transmission rate (∼2330 gm(-2)day(-1)), high elasticity (∼2...
October 13, 2016: Acta Biomaterialia
Eun-Ju Choi, Trishna Debnath, Yujiao Tang, Young-Bae Ryu, Sang-Ho Moon, Eun-Kyung Kim
BACKGROUND: Moringa oleifera (M. oleifera) is widely cultivated in tropical and subtropical regions and has been used as a vegetable and in traditional medicine. In this study, the anti-atopic dermatitis activity of the ethanol extract of M. oleifera leaf was investigated in vitro and in vivo. METHODS: For the in vitro study, HaCaT human keratinocytes were used for cytokines and MAPKinase assay. In the in vivo study, M. oleifera leaf ethanolic extract (MO) was topically applied to BALB/c mice with Dermatophagoides farinae extract (DFE; house dust mite extract)- and 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis (AD)...
October 12, 2016: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
Chen Tu, Shuang Wang, Xiao Hu, Wenju Wang, Yingying Dong, Shengxiang Xiao, Xiaopeng Wang
Bacterial infection is an important factor that can trigger and exacerbate psoriasis. The protein triggering receptor expressed on myeloid cells type-1 (TREM-1) is overexpressed in psoriasis and decreased after a successful treatment. Hypoxia inducible factor-1α (HIF-1α), subunit of the transcription factor HIF-1, has participated in angiogenesis and inflammation in psoriasis. Increased expressions of TREM-1 and HIF-1α are associated with the infection of microbial pathogens. However, the association between TREM-1 and HIF-1α still need to be elucidated...
October 15, 2016: Cell Biology International
Yojana R Chikate, Vishal V Dawkar, Ranjit S Barbole, Priyadarshini V Tilak, Vidya S Gupta, Ashok P Giri
Helicoverpa armigera is one of the major crop pests and is less amenable to current pest control approaches. RNA interference (RNAi) is emerging as a potent arsenal for the insect pest control over current methods. Here, we examined the effect on growth and development in H. armigera by targeting various enzymes/proteins such as proteases like trypsins (HaTry2, 3, 4 and 6), chymotrypsin (HaChy4) and cysteine protease like cathepsin (HaCATHL); glutathione S-transferases (HaGST1a, 6 and 8); esterases (HaAce4, HaJHE); catalase (HaCAT); super-oxide-dismutase (HaCu/ZnSOD); fatty acid binding protein (HaFabp) and chitin deacetylase (HaCda5b) through dsRNA approach...
October 2016: Pesticide Biochemistry and Physiology
Abderrakib Zahid, Julie Despres, Magalie Benard, Eric Nguema-Ona, Jerome Leprince, David Vaudry, Christophe Rihouey, Maité Vicré-Gibouin, Azeddine Driouich, Marie-Laure Follet-Gueye
Plant derived arabinogalactan proteins (AGP) were repeatedly confirmed as immunologically as well as dermatologically active compounds. However little is currently known regarding their potential activity towards skin innate immunity. Here, we extracted and purified AGP from acacia (Acacia senegal) and baobab (Adansonia digitata) seeds to investigate their biological effects on the HaCaT keratinocyte cell line in an in vitro system. While AGP from both sources did not exhibit any cytotoxic effect, AGP from acacia seeds enhanced cell viability Moreover, real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis showed that AGP extracted from both species induced a substantial overexpression of hBD-2, TLR-5, and IL1-α genes...
October 13, 2016: Journal of Cellular Physiology
J Wang, Y Qiu, N-W Shi, J-N Zhao, Y-C Wang, H Jiang, H-B Qian
OBJECTIVE: The aim of this study was to investigate the molecular mechanism into the keratinocyte migration, which is promoted by Transforming growth factor-β1 (TGF-β1) during wound healing. MATERIALS AND METHODS: In the present study, we investigated the regulation by TGF-β1 on phosphatase and tensin homolog (PTEN) expression, and microRNA-21 (miR-21) level with real-time quantitative PCR or/and Western blotting, and then examined the regulatory role of miR-21 on the PTEN expression and the mesenchymal transition, with real-time quantitative PCR, western blotting and luciferase reporter assay, and the migration of keratinocytic HaCaT cells with scratch assay...
September 2016: European Review for Medical and Pharmacological Sciences
Josefa P Alameda, Manuel Navarro, Ángel Ramírez, Angustias Page, Cristian Suárez-Cabrera, Rodolfo Moreno-Maldonado, Jesús M Paramio, M Carmen Fariña, Marcela Del Río, M Jesús Fernández-Aceñero, Ana Bravo, M Llanos Casanova
IKKα plays a mandatory role in keratinocyte differentiation and exerts an important task in non-melanoma skin cancer development. However, it is not fully understood how IKKα exerts these functions. To analyze in detail the role of IKKα in epidermal stratification and differentiation, we have generated tridimensional (3D) cultures of human HaCaT keratinocytes and fibroblasts in fibrin gels, obtaining human skin equivalents that comprise an epidermal and a dermal compartments that resembles both the structure and differentiation of normal human skin...
October 8, 2016: Oncotarget
Lizhao Guan, Yan Song, Jian Gao, Jianjun Gao, KeWei Wang
ANO1, a calcium-activated chloride channel, has been reported to be amplified or overexpressed in tissues of several cancers. However, reports on its roles in tumor progression obtained from cancer cell lines are inconsistent, suggesting that the role of ANO1 in tumorigenesis is likely dependent on either its expression level or cell-type expressing ANO1. To investigate the biological roles of ANO1 in different tumor cells, we, in this study, selected several cancer cell lines and a normal HaCaT cell line with high expression levels of ANO1, and examined the function of ANO1 in these cells using approaches of lentiviral knockdown and pharmacological inhibition...
October 8, 2016: Oncotarget
Ritesh K Srivastava, Changzhao Li, Zhiping Weng, Anupam Agarwal, Craig A Elmets, Farrukh Afaq, Mohammad Athar
Arsenicals are painful, inflammatory and blistering causing agents developed as chemical weapons in World War I/II. However, their large stockpiles still exist posing threat to public health. Phenylarsine oxide (PAO), a strong oxidant and a prototype arsenical is tested for its suitability to defining molecular mechanisms underlying arsenicals-mediated tissue injury. Topically applied PAO induces cutaneous erythema, edema and micro-blisters. These gross inflammatory responses were accompanied by the enhanced production of pro-inflammatory cytokines, ROS and unfolded protein response (UPR) signaling activation...
October 11, 2016: Scientific Reports
Jun Yeon Park, Jin Ho Kwak, Ki Sung Kang, Eun Bee Jung, Dong-Soo Lee, Sanghyun Lee, Yujung Jung, Ki Hyun Kim, Gwi Seo Hwang, Hye Lim Lee, Noriko Yamabe, Su-Nam Kim
ETHNOPHARMACOLOGICAL RELEVANCE: Jawoongo is a traditional drug ointment (with a traditional botanic formula) used for the treatment of burns and wounds in Korea. One of the components of Jawoongo is Lithospermi Radix (LR, the dried root of Lithospermum erythrorhizon Siebold & Zucc., also known as Zicao or Gromwell), which contains deoxyshikonin and its derivatives. OBJECTIVE: The aim of the present study was to investigate the effects of deoxyshikonin on wound healing...
October 7, 2016: Journal of Ethnopharmacology
Juthika Kundu, In Gyeong Chae, Kyung-Soo Chun
BACKGROUND: Fraxetin (7,8-dihydroxy-6-methoxy coumarin), a coumarin derivative, has been reported to possess antioxidative, anti-inflammatory and neuroprotective effects. A number of recent observations suggest that the induction of heme oxygenase-1 (HO-1) inhibits inflammation and tumorigenesis. In the present study, we determined the effect of fraxetin on HO-1 expression in HaCaT human keratinocytes and investigated its underlying molecular mechanisms. METHODS: Reverse transcriptase-PCR and Western blot analysis were performed to detect HO-1 mRNA and protein expression, respectively...
September 2016: Journal of Cancer Prevention
Martin Conda-Sheridan, Venkatareddy Udumula, Jennifer L Endres, Caleb N Harper, Lee Jaramillo, Haizhen A Zhong, Kenneth W Bayles
Community-associated methicillin resistant Staphylococcus aureus (CA-MRSA) has become a severe health concern because of its treatment difficulties. Herein, we report the synthesis and biological evaluation of two phenazine natural products and a series of phenazines that show promising activities against MRSA with MIC values in the low micromolar range. Basic studies revealed that these compounds are bacteriostatic agents. The most active compound also displayed promising IC50 values against HaCat cells. Finally, a QSAR model was developed to understand the key structural features of the molecules...
September 28, 2016: European Journal of Medicinal Chemistry
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