Metabolite Profile and Antiproliferative Effects in HaCaT Cells of a Salix reticulata Extract

Planta Medica ◽  
2017 ◽  
Vol 83 (14/15) ◽  
pp. 1149-1158 ◽  
Author(s):  
Elisabetta Corradi ◽  
Nadine Schmidt ◽  
Nathalie Räber ◽  
Maria De Mieri ◽  
Matthias Hamburger ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Migration ◽  
Time Lapse ◽  
Metabolite Profile ◽  
Brdu Incorporation ◽  
Dependent Manner ◽  
Hacat Cells ◽  
Meoh Extract ◽  
Concentration Dependent Manner ◽  
Phenolic Constituents

AbstractPhenolic constituents of Salix reticulata (Salicaceae) and antiproliferative activity of an extract and individual compounds were investigated in immortalized human non-tumorigenic keratinocytes (HaCaT). A MeOH extract from aerial parts afforded several flavonoids, including luteolin and apigenin glycosides (2–5 and 9) and catechin (1), two procyanidin fractions, and the phenolic glucosides picein (6), triandrin (7), and salicortin (8). In an adenosine triphosphate assay, the MeOH extract reduced cell viability by approximately 60 % at a concentration of 100 µg/mL. Cell proliferation was assessed with a BrdU incorporation ELISA assay. The extract inhibited proliferation of HaCaT cells in a concentration-dependent manner, with approximately 50 % inhibition at 100 µg/mL. In time-lapse assays, the extract showed distinct inhibitory effects on cell migration at concentrations of 12.5, 25, and 50 µg/mL. The activity of selected constituents was also determined. Luteolin-7-O-β-glucuronide (3) significantly inhibited cell proliferation at concentrations of 10 and 50 µM. In contrast, luteolin-7-O-β-glucopyranoside (2) and a procyanidin fraction (P1) had only weak effects, while picein (6) and salicortin (8) did not affect cell proliferation. Luteolin-7-O-β-glucuronide (10 µM) and, to a lesser extent, the procyanidin fraction (10 µg/mL) also inhibited cell migration.

Antiproliferative Effects of Pterodon pubescens Extract and Isolated Diterpenes in HaCaT Cells

Planta Medica ◽  
2020 ◽  
Author(s):  
Rosanna Tarkany Basting ◽  
Ilza Maria de Oliveira Sousa ◽  
Veronika Butterweck ◽  
Mary Ann Foglio
Keyword(s):  
Cell Migration ◽  
Methyl Ester ◽  
Local Treatment ◽  
Time Lapse ◽  
Molecular Structures ◽  
Brdu Incorporation ◽  
Hacat Cells ◽  
Antiproliferative Effects ◽  
Anti Inflammatory ◽  
Incorporation Assay

Abstract Pterodon pubescens fruits are popularly used because of their analgesic and anti-inflammatory actions, which are attributed to the isolated compounds with a vouacapan skeleton. This work aimed to evaluate the antiproliferative and anti-inflammatory effects of a P. pubescens fruit dichloromethane extract and the vouacapan diterpene furan isomerʼs mixture (1 : 1) (6α-hydroxy-7β-acetoxy-vouacapan-17β-oate methyl ester and 6α-acetoxy-7β-hydroxy-vouacapan-17β-oate methyl ester isomers) in HaCaT cells using the cell migration and the BrDU incorporation assay. Levels of IL-8 were measured by ELISA after TNF-α stimulation. HPLC/DAD analysis of the extract revealed the expressive presence of vouacapan diterpene furan isomerʼs mixture. P. pubescens extract (1.5625 – 25 µg/mL) and vouacapan diterpene furan isomerʼs mixture (3.125 – 50 µM) inhibited cell proliferation as indicated by a decreased BrdU-incorporation. For the evaluation of cell migration, time-lapse microscopy was used. P. pubescens presented inhibition on cell migration at all concentrations tested (3.125 – 12.5 µg/mL), whereas for the VDFI mixture, the inhibition was only observed at the highest concentrations (12.5 and 25 µM) tested. Furthermore P. pubescens extract and vouacapan diterpene furan isomerʼs mixture significantly decreased IL-8 levels. Our results showed antiproliferative and anti-inflammatory effects on HaCaT cells treated with the extract and the vouacapan isomerʼs mixture, without affecting cell viability. These activities could be attributed to the voucapan molecular structures. In conclusion, topical products developed of P. pubescens extract or the voucapan isomerʼs mixture should be further studied as a potential product for local treatment against hyperproliferative lesions as in psoriasis vulgaris, representing an alternative treatment approach.

scholarly journals ALK Inhibitors-Induced M Phase Delay Contributes to the Suppression of Cell Proliferation

Cancers ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 1054
Author(s):  
Sirajam Munira ◽  
Ryuzaburo Yuki ◽  
Youhei Saito ◽  
Yuji Nakayama
Keyword(s):  
Cell Proliferation ◽  
Spindle Assembly Checkpoint ◽  
Time Lapse ◽  
Phase Delay ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Alk Inhibitors ◽  
Anaplastic Lymphoma ◽  
Monopolar Spindle ◽  
M Phase

Anaplastic lymphoma kinase (ALK), a receptor-type tyrosine kinase, is involved in the pathogenesis of several cancers. ALK has been targeted with small molecule inhibitors for the treatment of different cancers, but absolute success remains elusive. In the present study, the effects of ALK inhibitors on M phase progression were evaluated. Crizotinib, ceritinib, and TAE684 suppressed proliferation of neuroblastoma SH-SY5Y cells in a concentration-dependent manner. At approximate IC50 concentrations, these inhibitors caused misorientation of spindles, misalignment of chromosomes and reduction in autophosphorylation. Similarly, knockdown of ALK caused M phase delay, which was rescued by re-expression of ALK. Time-lapse imaging revealed that anaphase onset was delayed. The monopolar spindle 1 (MPS1) inhibitor, AZ3146, and MAD2 knockdown led to a release from inhibitor-induced M phase delay, suggesting that spindle assembly checkpoint may be activated in ALK-inhibited cells. H2228 human lung carcinoma cells that express EML4-ALK fusion showed M phase delay in the presence of TAE684 at about IC50 concentrations. These results suggest that ALK plays a role in M phase regulation and ALK inhibition may contribute to the suppression of cell proliferation in ALK-expressing cancer cells.

Tripterine inhibits proliferation, migration and invasion of breast cancer MDA-MB-231 cells by up-regulating microRNA-15a

2019 ◽  
Vol 400 (8) ◽  
pp. 1069-1078 ◽  
Author(s):  
Anjun Zuo ◽  
Peng Zhao ◽  
Yu Zheng ◽  
Hui Hua ◽  
Xingang Wang
Keyword(s):  
Breast Cancer ◽  
Cell Proliferation ◽  
Annexin V ◽  
Cell Counting ◽  
Brdu Incorporation ◽  
Migration And Invasion ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Apoptosis Detection ◽  
Incorporation Assay

Abstract Breast cancer is the most commonly diagnosed cancer in women worldwide. Tripterine is an important active component isolated from Triperygium wilfordii Hook F. This study investigated the effects of tripterine on breast cancer cell proliferation, migration, invasion and apoptosis, as well as microRNA-15a (miR-15a) expression. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed to measure the expression of miR-15a. Cell transfection was conducted to change the expression of miR-15a. Viability, proliferation, migration, invasion and apoptosis of MDA-MB-231 cells were assessed using the cell counting kit-8 (CCK-8) assay, BrdU incorporation assay, Annexin V-FITC/PI apoptosis detection kit and two-chamber Transwell assay, respectively. Expression of key factors involving in cell proliferation, migration, invasion and apoptosis, as well as the PI3K/AKT and JNK pathways, were evaluated using Western blotting. We found that tripterine inhibited MDA-MB-231 cell viability, proliferation, migration and invasion, but induced cell apoptosis. Moreover, tripterine up-regulated the expression of miR-15a in a concentration-dependent manner and miR-15a participated in the effects of tripterine on MDA-MB-231 cell proliferation, migration, invasion and apoptosis. In addition, tripterine inactivated PI3K/AKT and JNK pathways in MDA-MB-231 cells by up-regulating miR-15a. In conclusion, tripterine inhibited proliferation, migration and invasion of breast cancer MDA-MB-231 cells by up-regulating miR-15a and inactivating PI3K/AKT and JNK pathways.

Saffron Flower Extract Promotes Scratch Wound Closure of Keratinocytes and Enhances VEGF Production

Planta Medica ◽  
2017 ◽  
Vol 83 (14/15) ◽  
pp. 1176-1183 ◽  
Author(s):  
Sheela Verjee ◽  
Eliane Garo ◽  
Sarah Pelaez ◽  
Orlando Fertig ◽  
Matthias Hamburger ◽  
...  
Keyword(s):  
Cell Migration ◽  
Wound Closure ◽  
Value Added ◽  
Acetone Extract ◽  
Crocus Sativus ◽  
Brdu Incorporation ◽  
Dependent Manner ◽  
Hacat Cells ◽  
Tnf Α ◽  
Scratch Wound

AbstractDuring saffron (Crocus sativus) spice production, large amounts of floral biowaste are generated. It was the aim of this study to develop a value-added product from saffron floral biowaste to be used as a natural cosmetic ingredient. HPLC-PDA-MS analysis of saffron flower extracts revealed the presence of flavonols with the highest amounts in the acetone extract. Kaempferol-3-O-sophoroside was identified as the main flavonoid in the acetone extract (saffron flower acetone extract). Saffron flower acetone extract and kaempferol-3-O-sophoroside were tested in HaCaT cells for potential effects on cell migration, proliferation, and for anti-inflammatory properties. Saffron flower acetone extract concentration dependently (50–200 µg/mL) augmented cell proliferation, as indicated by an increased BrdU-incorporation, while kaempferol-3-O-sophoroside (1–50 µM) had no effect. Furthermore, treatment of HaCaT cells with saffron flower acetone extract, but not with kaempferol-3-O-sophoroside, concentration-dependently increased vascular endothelial growth factor secretion (control 49.72 pg/mL vs. saffron flower acetone extract at 200 µg/mL 218.60 pg/mL). Cell migration was determined using time-lapse microscopy and a modification of the scratch-wound assay in which saffron flower acetone extract significantly improved wound closure compared to the untreated control. Overproduction of the proinflammatory cytokines interleukin-8 and interleukin-6 in HaCaT cells was induced by TNF-α. Kaempferol-3-O-sophoroside (10–50 µM), but not saffron flower acetone extract, inhibited TNF-α-induced IL-8 secretion. The effect was comparable to 10 µM hydrocortisone (positive control). Interestingly, saffron flower acetone extract further increased IL-6 levels in TNF-α-treated HaCaT cells in a concentration-dependent manner. In summary, the pronounced wound healing properties of saffron flower acetone extract present a promising application for the cosmetic industry.

scholarly journals Complex effects of kinase localization revealed by compartment-specific regulation of protein kinase A activity

2021 ◽  
Author(s):  
Rebecca LaCroix ◽  
Benjamin Lin ◽  
Andre Levchenko
Keyword(s):  
Cell Migration ◽  
Protein Kinase ◽  
Protein Kinase A ◽  
Kinase Activity ◽  
Single Cells ◽  
Regulatory Subunit ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Specific Regulation ◽  
Kinase A

SummaryKinase activity in signaling networks frequently depends on regulatory subunits that can both inhibit activity by interacting with the catalytic subunits and target the kinase to distinct molecular partners and subcellular compartments. Here, using a new synthetic molecular interaction system, we show that translocation of a regulatory subunit of the protein kinase A (PKA-R) to the plasma membrane has a paradoxical effect on the membrane kinase activity. It can both enhance it at lower translocation levels, even in the absence of signaling inputs, and inhibit it at higher translocation levels, suggesting its role as a linker that can both couple and decouple signaling processes in a concentration-dependent manner. We further demonstrate that superposition of gradients of PKA-R abundance across single cells can control the directionality of cell migration, reversing it at high enough input levels. Thus complex in vivo patterns of PKA-R localization can drive complex phenotypes, including cell migration.

scholarly journals 1289. Pravibismane is a Potent, Broad Spectrum Anti-Infective Small Molecule that Rapidly Disrupts Bacterial Bioenergetics and Halts Bacterial Growth

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S659-S660
Author(s):  
Brett Baker
Keyword(s):  
Membrane Potential ◽  
Cell Growth ◽  
Protein Expression ◽  
Broad Spectrum ◽  
Time Lapse ◽  
Luciferase Assay ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Time Lapse Microscopy ◽  
Atp Levels

Abstract Background The rise in resistance to existing antimicrobials has prompted a need for the development of novel antibiotics. Microbion has identified a novel compound, pravibismane, with potent broad spectrum anti-infective and anti-biofilm activity. Methods Here we used a variety of assays, including Bacterial Cytological Profiling (BCP), to analyze pravibismane in E.coli to gain insight into its likely mechanism of action (MOA). The BCP profile of pravibismane suggested it rapidly shut down cell growth, potentially by turning off cellular gene or protein expression. This was confirmed using a plasmid based GFP induction assay in E.coli tolC that showed pravibismane strongly reduced expression of GFP. The kinetics, reversibility and MOA of pravibismane was further characterized by using time-lapse microscopy, wash out experiments and measurements of both membrane potential and relative intracellular ATP levels. Results We found that pravibismane acts rapidly (within 30 mins) to completely halt cell growth rather than causing immediate cell lysis such as that observed with non-specific cell damaging agents bleach or detergent. Inhibitor wash out experiments in which cells were exposed to pravibismane for 2 hours, washed to remove the compound, and then observed using time-lapse microscopy revealed that the effect of pravibismane is reversible and that cells recovered 8-12 hrs after removing the compound. Wash out experiments with an E.coli tolC strain carrying a plasmid with an IPTG inducible GFP demonstrated that transcription and translation ultimately resumed in most cells after washout. The bioenergetics of the membrane was measured using DiBAC 4(5), a membrane potential sensitive dye which can enter depolarized cells, which revealed that pravibismane caused depolarization of the membrane within 30 mins of exposure in a concentration dependent manner. Finally, a luciferase assay determined pravibismane reduced ATP levels (resulting in decreased luminescence) within 15 mins of exposure in a concentration dependent manner unlike antibiotic controls that had modest or no effect on luminescence. Conclusion Our results suggest that pravibismane acts rapidly to disrupt cellular bioenergetics, resulting in the immediate cessation of cell growth and protein expression. Disclosures Brett Baker, M.Sc., D.C., Microbion Corporation (Board Member, Employee)

scholarly journals Betulinic Acid Suppresses Ovarian Cancer Cell Proliferation through Induction of Apoptosis

Biomolecules ◽  
2019 ◽  
Vol 9 (7) ◽  
pp. 257 ◽  
Author(s):  
Dahae Lee ◽  
Seoung Rak Lee ◽  
Ki Sung Kang ◽  
Yuri Ko ◽  
Changhyun Pang ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Cancer Cell ◽  
Betulinic Acid ◽  
Molecular Mechanisms ◽  
Underlying Mechanism ◽  
Nuclear Staining ◽  
Dependent Manner ◽  
Meoh Extract ◽  
Concentration Dependent Manner ◽  
Induction Of Apoptosis

Ovarian cancer is one of the leading causes of cancer deaths worldwide in women, and the most malignant cancer among the different gynecological cancers. In this study, we explored potentially anticancer compounds from Cornus walteri (Cornaceae), the MeOH extract of which has been reported to show considerable cytotoxicity against several cancer cell lines. Phytochemical investigations of the MeOH extract of the stem and stem bark of C. walteri by extensive application of chromatographic techniques resulted in the isolation of 14 compounds (1–14). The isolated compounds were evaluated for inhibitory effects on the viability of A2780 human ovarian carcinoma cells and the underlying molecular mechanisms were investigated. An 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was employed to assess the anticancer effects of compounds 1–14 on A2780 cells, which showed that compound 11 (betulinic acid) reduced the viability of these cells in a concentration-dependent manner and had an half maximal (50%) inhibitory concentration (IC50) of 44.47 μM at 24 h. Nuclear staining and image-based cytometric assay were carried out to detect the induction of apoptosis by betulinic acid. Betulinic acid significantly increased the condensation of nuclei and the percentage of apoptotic cells in a concentration-dependent manner in A2780 cells. Western blot analysis was performed to investigate the underlying mechanism of apoptosis. The results indicated that the expression levels of cleaved caspase-8, -3, -9, and Bax were increased in A2780 cells treated with betulinic acid, whereas those of Bcl-2 were decreased. Thus, we provide the experimental evidence that betulinic acid can induce apoptosis in A2780 cells through both mitochondria-dependent and -independent pathways and suggest the potential use of betulinic acid in the development of novel chemotherapeutics for ovarian cancer therapy.

scholarly journals TRB3 is elevated in psoriasis vulgaris lesions and mediates HaCaT cells proliferation in vitro

2017 ◽  
Vol 65 (7) ◽  
pp. 1084-1088 ◽  
Author(s):  
Xiao-Jing Yu ◽  
Tie-Jun Song ◽  
Lu-Wei Zhang ◽  
Ying Su ◽  
Ke-Yu Wang ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Messenger Rna ◽  
Psoriasis Vulgaris ◽  
Metabolic Diseases ◽  
Brdu Incorporation ◽  
Cell Cycle Distribution ◽  
Hacat Cells ◽  
Keratinocyte Proliferation

Psoriasis is a chronic skin disease characterized by abnormal keratinocyte proliferation and differentiation, inflammation, and angiogenesis. Overexpression of tribbles homolog3 (TRB3), which belongs to the tribbles family of pseudokinases, has been found in several human tumors and metabolic diseases, but its role in psoriasis has not been fully clarified. The aim of this study is to investigate the expression of TRB3 in psoriasis and explore its roles in the proliferation of keratinocytes. Twenty-four patients with psoriasis vulgaris were recruited for the study. Diagnosis of psoriasis was based on clinical and histologic examinations. Immunohistochemistry and real-time reverse transcription PCR (RT-PCR) were performed to determine protein and messenger RNA (mRNA) expression of TRB3 in psoriasis lesions. 5-Bromo-2-deoxyUridine (BrdU) incorporation assay were performed for cell proliferation. Cell cycle distribution was assessed by flow cytometry analysis. The levels of TRB3 is elevated in psoriatic lesions compared with psoriatic non-lesions. The HaCat cells expressed the TRB3 gene. We found TRB3 silencing to significantly inhibit HaCat cell proliferation. Furthermore, the specific knockdown of TRB3 slowed down the cell cycle at the gap 0/first gap phase. In conclusion, our data suggest that TRB3 is overexpressed in lesions of patients with psoriasis and may be involved in the abnormal proliferation of keratinocytes. Therefore, TRB3 may be a potential therapeutic target for psoriasis.

scholarly journals Metformin Suppressed CXCL8 Expression and Cell Migration in HEK293/TLR4 Cell Line

2017 ◽  
Vol 2017 ◽  
pp. 1-11 ◽  
Author(s):  
Zhihui Xiao ◽  
Wenjun Wu ◽  
Vladimir Poltoratsky
Keyword(s):  
Cell Proliferation ◽  
Cell Migration ◽  
Tumor Cell ◽  
Nuclear Translocation ◽  
High Dose ◽  
Dependent Manner ◽  
Tumor Cell Proliferation ◽  
Cell Proliferation And Migration ◽  
And Migration ◽  
Proliferation And Migration

Chronic inflammation is associated with cancer. CXCL8 promotes tumor microenvironment construction through recruiting leukocytes and endothelial progenitor cells that are involved in angiogenesis. It also enhances tumor cell proliferation and migration. Metformin, type II diabetes medication, demonstrates anticancer properties via suppressing inflammation, tumor cell proliferation, angiogenesis, and metastasis. This study intended to address the role of metformin in regulation of CXCL8 expression and cell proliferation and migration. Our data indicated that metformin suppressed LPS-induced CXCL8 expression in a dose-dependent manner through inhibiting NF-κB, but not AP-1 and C/EBP, activities under the conditions we used. This inhibitory effect of metformin is achieved through dampening LPS-induced NF-κB nuclear translocation. Cell migration was inhibited by metformin under high dose (10 mM), but not cell proliferation.

scholarly journals Evaluation of Anti-Inflammatory Activities of Qingre-Qushi Recipe (QRQS) against Atopic Dermatitis: Potential Mechanism of Inhibition of IL-33/ST2 Signal Transduction

2017 ◽  
Vol 2017 ◽  
pp. 1-11 ◽  
Author(s):  
Mengjiao Chen ◽  
Peijun Ding ◽  
Lili Yang ◽  
Xufeng He ◽  
Chunjie Gao ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Molecular Mechanisms ◽  
Histological Study ◽  
High Dose ◽  
Dependent Manner ◽  
Hacat Cells ◽  
Concentration Dependent Manner ◽  
Dermal Thickness ◽  
Significant Difference ◽  
Anti Inflammatory

To evaluate the anti-inflammatory activities of QRQS against AD and the inhibitory molecular mechanisms of IL-33/ST2 signal transduction, BALB/c mice were divided into six groups (normal control, OVA control, low-dose of QRQS, middle-dose of QRQS, high-dose of QRQS, and cetirizine) and epicutaneously exposed to ovalbumin or PBS for 3 weeks and treated with QRQS for 2 weeks. Skin biopsies and blood samples were obtained for histological study, antibody analysis, and RNA isolation. HaCaT cells, stimulated by TNF-α and IFN-γ, were treated with QRQS to evaluate mRNA and protein expression by RT-PCR and ELISA. QRQS decreased both epidermal and dermal thickness, alleviated dermatitis, and reduced IL-33 and ST2 positive cell numbers. The concentration of specific IgE, IgG, IgG1, and IgG2a antibodies in serum and the expression of IL-33, ST2, IL-1RAcP, IL-4, and IL-13 mRNA in the skin were suppressed. No significant difference exists in TNF-α or IFN-γ. QRQS decreased IL-33 mRNA and protein secretion in HaCaT cells exposed to TNF-α and IFN-γ in a time- and concentration-dependent manner. QRQS regulates related molecule expression of ovalbumin-induced dermatitis involved in the IL-33/ST2 signaling axis in the treatment of acute AD.

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