Heterocyclic aromatic compounds from Artemisia judaica (L.) induced caspase-3 mediated apoptosis in metastatic breast cell lines

Extensive uses of calcium titanate nanoparticles (CaTiO3-NPs) and erbium oxide nanoparticles (Er2O3-NPs) increase their release into the environment and human exposure, particularly through skin contact. However, there are almost no studies available on the effect of these nanoparticles on skin integrity. Therefore, this study was undertaken to estimate CaTiO3-NP- or Er2O3-NP-induced cytotoxicity and genotoxicity in normal human skin fibroblast (HSF) cells. Cell viability was measured using sulforhodamine B (SRB) assay, while the level of DNA damage was detected using the alkaline comet assay. The intracellular levels of reactive oxygen species (ROS) as well as the expression level of p53, Bax, and Bcl2 genes were detected. Although the viability of HSF cells was non-markedly changed after 24 h, prolonged treatment with CaTiO3-NPs or Er2O3-NPs for 72 h induced concentration-dependent death of HSF cells. Treatment of normal HSF cells with IC50/72 h of CaTiO3-NPs or Er2O3-NPs did not cause marked changes in the intracellular level of ROS, DNA damage parameters, and expression levels of apoptosis genes compared to their values in the untreated HSF cells. We thus concluded that CaTiO3-NPs or Er2O3-NPs cause time- and concentration-dependent cytotoxicity toward normal HSF cells. However, safe and non-genotoxic effects were demonstrated by the apparent non-significant changes in intracellular ROS level, DNA integrity, and apoptotic genes’ expression after exposure of normal HSF cells to nanoparticles. Thus, it is recommended that further studies be conducted to further understand the toxic and biological effects of CaTiO3-NPs and Er2O3-NPs.

To study the impact of small interference RNA (siRNA) targeting cyclin A2 gene on the growth of MG-63 and HSF cells and to explore whether cyclin A2 siRNAs could become a useful tool in the treatment of osteosarcoma. One pair of siRNA targeting the cyclin A2 mRNA and a pair of nonsense siRNA were designed according the current criteria. SiRNA was chemically synthesized and purified. The siRNA was transfected into osteosarcoma cell line MG-63 and normal human skin fibroblast (HSF) cells via oligofectamine. Cells transfected with nonsense siRNA served as the negative control and those only treated with PBS as the blank control group. Quantitative fluorescence RT-PCR, Western-blot, MTT assay, reverse transcriptase (RT)-PCR, flow cytometry and colony-forming test were employed to evaluate the efficacy of RNA interference. At the same time, the mRNA expression of PCNA and cyclin B1 in siRNA treated MG-63 cells were examined. 1 nmol/L, 10 nmol/L, 50 nmol/L and 100 nmol/L cyclin A2-siRNA can reduced cyclin A2 mRNA and protein expression respectively by 9.43%, 56.35%, 79.17% and 84.30% as compared with that of the blank control group, whereas the negative and blank control groups had similar expression levels. After 48 h treatment with 10 nmol/L siRNA, MG-63 cells were arrested in G0/G1 phase and the proliferation of this tumor cell was suppressed by 39.06% 48 h after transfection. Furthermore, the treated MG-63 cells showed less colony-forming ability. Increasing the siRNA concentration to 50 nmol/L can further inhibit the proliferation of MG-63 cells by 54.94%. In addition, the cyclin A2-depleted MG-63 cells showed decreased levels of PCNA and cyclin B1. In contrast, although cyclin A2 mRNA and protein expression in HSF reduced 58.13% 48 h after treatment by 50 nmol/L siRNA, these cells exhibited only a slight change in cell cycle, and no clear inhibition of proliferation and impaired plate colony-forming ability was observed. Cyclin A2 gene maybe served as a potential target for tumor therapy. RNA interference induces obvious inhibition of cyclin A2 mRNA and protein expression in MG-63 and HSF cells, which consequently downregulate the proliferation of MG-63 cells. There is few inhibitory effect on the proliferation by siRNAs for HSF cells. These results indicate that siRNAs against cyclin A2 could become a potential antiproliferative tool in future antitumor therapy.

This study was designed to assess the effect of brentuximab vedotin on several breast cancer cell lines in terms of promoting apoptosis and managing cancer progression. Additionally, the study investigated the potential of repurposing this drug for new therapeutic reasons, beyond its original indications. The study evaluates the cytotoxic effects of Brentuximab vedotin across five cell lines: normal human skin fibroblasts (HSF), three breast cancer cell lines (MCF-7, MDA-MB-231, and T-47D), and histiocytic lymphoma (U-937). Brentuximab treatment was administered at four time points (0, 24, 48, and 72 h), with cell viability assessed at each interval. HSF cells, serving as controls, exhibited minimal viability loss (above 70%), indicating limited toxicity in normal fibroblasts. In contrast, MCF-7 and MDA-MB-231 cells demonstrated time-dependent reductions in viability, with a pronounced decline by 72 h, suggesting Brentuximab’s efficacy in both ER-positive and triple-negative breast cancer. T-47D cells also showed decreased viability, though at a slower rate. U-937 cells exhibited the most substantial reduction, highlighting Brentuximab’s potent activity against hematologic malignancies. Wound healing assays further revealed that Brentuximab significantly impaired the migration and healing capacity of cancer cells compared to untreated controls. Additionally, cell cycle analysis indicated G2/M phase arrest in cancer cells, particularly in MCF-7 and MDA-MB-231, while HSF cells remained largely unaffected. Apoptosis detection confirmed Brentuximab-induced cell death, with significant increases in late apoptosis in cancer lines, especially by 72 h. Gene expression analysis revealed upregulation of pro-apoptotic genes (BAX, Caspase 3, and Caspase 9) in cancer cells, alongside a decrease in anti-apoptotic BCL-2 expression. These findings suggest Brentuximab’s selective cytotoxicity against cancer cells and its potential as an effective therapeutic agent, particularly in breast cancer and histiocytic lymphoma.

Background: A wide use of Silver Nanoparticles (AgNPs) in consumer products, industry, pharmaceuticals, and medical sciences, including dermatology, has always caused concerns related to its adverse effects on the environment and human safety. Nano-sized particles can be uptaken by the body through various routes, especially the skin, which is the widest organ and the first contact surface for different nanomaterials and other foreign agents. Objectives: The aim of this investigation was to assess the viability of a vulnerable normal skin cell line to cytotoxicity of AgNPs with the same effective size and concentrations used for various microbial agents, such as Candida species. Methods: Synthesized AgNPs were characterized using UV-visible absorption spectroscopy, X-ray diffraction, Dynamic Light Scattering (DLS), and Transmission Electron Microscopy (TEM). Different concentrations of the aqueous colloidal AgNPs (2.17 to 69.5 µg/mL) were tested on normal Human Skin Fibroblast (HSF) cell line using MTT assay after 24 and 48 hours of exposure. Results: The analysis demonstrated that the synthesized AgNPs were mainly spherical with an average size of 6.03 nm. The MTT findings showed a significant concentration- and time-dependent manner in the reduction of cell viability. The calculated IC50 values were 30.64 and 14.98 µg/mL for 24 and 48 hours of incubation, respectively. Conclusions: The cytotoxic effect of AgNPs was higher for Human Skin Fibroblast cells after 48 hours of exposure compared with 24 hours. According to the observed results, small nano-sized silver particles could potentially have cytotoxic effects on normal cells, especially at high concentrations and prolonged exposure times.

Dose-dependent proliferative and cytotoxic effects of melatonin on human epidermoid carcinoma and normal skin fibroblast cells

Formononetin alleviates thermal injury-induced skin fibroblast apoptosis and promotes cell proliferation and migration

Adenoviral vectors with E1A regulated by tumor-specific promoters are selectively cytolytic for breast cancer and melanoma.

Effect of low intensity laser interaction with human skin fibroblast cells using fiber-optic nano-probes

Infected wound is one of the most common and serious problem in wound management. Cold atmospheric plasma (CAP) is considered to have a good effect in wound healing as a new type medicine. However, there is a key issue that has not been addressed in the treatment of infected wounds by plasma. Bacteria are always found in the deep region of the wound. When plasma is used to treat wounds, it also acts on normal tissue cells while decontaminating. What is the difference between the same dose of plasma acting on bacteria and normal cells? In this study, the most common bacteria (S. aureus, P. aeruginosa, and E. coli) in infected wound and two kinds of normal skin cells (human keratinocyte and human skin fibroblasts (HSF)) were selected to study the difference of the effects of the same dose of plasma on bacteria and cells. The results reveal that three kinds of 106 CFU mL bacteria could be effectively inactivated by 5 order after plasma treatment 3 min, and P. aeruginosa was more sensitive to plasma (could be inactivated 5 order after 2 min treatment). The 104 mL keratinocyte and HSF were treated with the same dose of plasma; keratinocyte can maintain over 90% of the activity and HSF cells can maintain over 70% of the activity. Moreover, the level of collagen I secreted by HSF increased. Therefore, cells can remain a high activity when a plasma dose capable of inactivating bacteria is applied to them.

Differential DNA replication origin activities in human normal skin fibroblast and HeLa cell lines

Nucleoside-catabolizing enzyme activities in primary rabbit kidney cells and human skin fibroblasts

PHLPP2 (PH domain leucine-rich repeat protein phosphatase 2) terminates Akt and protein kinase C (PKC) activity by specifically dephosphorylating these kinases at a key regulatory site, the hydrophobic motif (Ser-473 in Akt1). Here we identify a polymorphism that results in an amino acid change from a Leu to Ser at codon 1016 in the phosphatase domain of PHLPP2, which reduces phosphatase activity toward Akt both in vitro and in cells, in turn resulting in reduced apoptosis. Depletion of endogenous PHLPP2 variants in breast cancer cells revealed the Ser-1016 variant is less functional toward both Akt and PKC. In pair-matched high grade breast cancer samples we observed retention of only the Ser allele from heterozygous patients (identical results were observed in a pair-matched normal and tumor cell line). Thus, we have identified a functional polymorphism that impairs the activity of PHLPP2 and correlates with elevated Akt phosphorylation and increased PKC levels.

BackgroundPan-drug resistance (PDR) is a ticking time bomb, as it causes high human hospitalizations and mortality rates. For instance, Pseudomonas aeruginosa is associated with significant rates of urinary tract infections (UTIs) due to several reasons including antibiotic resistance, biofilm formation and the presence of various virulence factors. Consequently, there is an urgent need for safe and effective alternative antibacterials. Phage therapy is a promising alternative that uses naturally occurring bacteriophages (phages). Therefore, our present study investigated the isolation and characterization of a novel virulent phage (vB_Ps_ZCPS13) against the PDR Pseudomonas aeruginosa strain (Ps13).MethodsPhage vB_Ps_ZCPS13 was isolated from raw sewage water in Egypt during the springtime. The isolated phage was purified and amplified, followed by estimating its purity and genome size using pulsed-field gel electrophoresis (PFGE), morphology using transmission electron microscopy (TEM), antibacterial activity against other P. aeruginosa hosts, physiochemical stability studies, whole genome sequencing, antibiofilm activity on urinary catheters using scanning electron microscopy (SEM), and cytotoxicity assays against normal human skin fibroblast (HSF) cell lines.ResultsBased on vB_Ps_ZCPS13 morphology under TEM, the phage has been classified as a myovirus. In consistent with the PFGE results, DNA sequencing revealed a phage genome size of 92,443 bp, with lytic-associated genes and no antimicrobial resistance or virulence factors. Phage vB_Ps_ZCPS13 presented a wide host range of over 93% of tested clinical isolates having different multiple antibiotic resistance (MAR) indices. Furthermore, phage vB_Ps_ZCPS13 exhibited high efficiency in plaque formation (EOP ≥ 1) against 13% of the strains and exhibited low frequencies of bacteriophage insensitive mutants (BIM). The physical stability test against harsh environmental conditions revealed phage stability within a pH range of 3.0–11.0 and stable at temperatures below 70 °C. Phage vB_Ps_ZCPS13 also exposed a significant antibacterial activity in vitro across different MOIs, with the highest reduction in bacterial growth observed at lower MOIs. Furthermore, vB_Ps_ZCPS13 demonstrated potent biofilm inhibition and clearance capabilities, effectively eradicating P. aeruginosa from the urinary catheter surface. Moreover, the phage presented no cytotoxicity against normal human skin fibroblast (HSF) cell lines at high titer.ConclusionsOur study offers an effective phage as a therapeutic candidate against PDR Gram-negative P. aeruginosa infections, including catheter-associated urinary tract infections.

Abstracts Allergic inflammation is thought to be a Th2 cell‐dominant immune response during which tissue‐resident fibroblasts produce chemokines which contribute to the recruitment of migratory leukocytes to sites of tissue injury. Thymus and activation‐regulated chemokine (TARC; CCL17) is a potent member of the CC chemokine family and a selective chemoattractant for Th2 cells. In order to study the regulatory profiles of TARC production by TNF‐α, IFN‐γ, and IL‐4 in human normal skin fibroblast, CCD‐986sk cell line was used. The expression of TARC protein was measured using ELISA, and mRNA level was detected by RT‐PCR. The combination of TNF‐α and IL‐4 induced a time‐ and dose‐dependent synergistic increase in the expression of TARC at both protein and mRNA levels in the cultured human skin fibroblasts. Exposure of the cells to single cytokine had no effect on TARC expression. The high concentration (100 ng/ml) and long incubation time (72 h) of IFN‐γ further enhanced the TARC production induced by TNF‐α/IL‐4 in the skin fibroblast. This synergistic effect of Th1 and Th2 type cytokines on TARC production by skin fibroblasts may contribute to the inflammatory cell infiltration and tissue damage with allergic inflammation. Keywords: Allergic inflammationskin fibroblast cellthymus and activation‐regulated chemokine (TARC;CCL17)CCD‐986sk cell Notes To whom correspondence should be addressed. Tel: +82–42–280–2432; Fax: +82–42–285–2432 E‐mail address: chyun@dju.ac.kr

This study is an investigation into the inhibitory effect of seawater pearl hydrolysate (SPH) on the UVA-induced photoaging of human skin fibroblast (HSF) cells, and the mechanism thereof. HSF cells were cultured and irradiated with a UVA 0–50 J·cm−2 dose gradient. The cell inhibition rate was detected using the CCK8 method, and the half-inhibitory dose was determined. Based on this, the dose of UVA irradiation for the follow-up experiment was selected to establish a photoaging model of the HSF cells. The cells were divided into a normal (N) group, UVA-irradiated (UVA) group, SPH low dose (SPHL) group, SPH medium dose (SPHM) group, and SPH high dose (SPHH) group. The photoaging model of HSF cells was established by UVA irradiation in the UVA, SPHL, SPHM, and SPHH groups; the SPHL, SPHM, and SPHH groups were treated with SPH at concentrations of 50, 100, and 200 mg·L−1, respectively, at the same time. After 24 and 48 h of culture, the reactive oxygen species (ROS) level of the HSF cells was detected by flow cytometry, and the required culture time of the HSF cells for the follow-up experiment was selected. The malondialdehyde and glutathione contents, as well as the activities of the superoxide dismutase, catalase, and glutathione peroxidase in the HSF cells, were detected by biochemical methods. The levels of expression of MMP-1 and collagen I protein in HSF cells were detected by the western blot test, the extent of aging of HSF cells was detected by β-galactosidase staining, and the apoptosis level of HSF cells was detected by flow cytometry. The results show that SPH inhibits the UVA-induced photoaging of HSF cells in a dose-dependent manner within a certain concentration range, and the effect of a concentration of 200 mg·L–1 was the most significant. The mechanism is related to improving the antioxidant activity of photoaging HSF cells to eliminate excessive ROS. It can inhibit apoptosis, reduce the protein expression of MMP-1, and effectively control the degradation of collagen I protein in photoaging HSF cells. Therefore, SPH offers potential for use in sunscreen cosmetics.