(1) Background: The purpose of this study was to investigate the effects of low-dose non-thermal atmospheric plasma (NTP) on the proliferation and migration of human immortalized keratinocytes (HaCaT cells) and its molecular mechanisms. (2) Methods: The effects of NTP on HaCaT cells were detected by cell viability, wound healing assay, cell cycle, mitochondrial membrane potential detection, and western blot, and the role of reactive oxygen species (ROS) content in low-dose NTP was explored. (3) Results: Results showed that the proportion of cells in G1-S phase transition, cell migration ability, and the expression of Cyclin D1 and STAT3 reached the peak at 10 s treatment group, while the cell viability and the expression of PI3K, AKT, mTOR, ERK, WNT, and β-catenin proteins reached the peak at 15 s treatment group. (4) Conclusions: These results manifested that ROS produced by low-dose NTP promoted the proliferation of HaCaT cells by activating the PI3K/AKT/mTOR signaling pathway and also promoted the migration by activating the WNT/β-catenin signaling pathway. Therefore, these results will be useful for the application of low-dose NTP in the treatment of wound healing.
Low-temperature plasma (LTP) is an emerging biomedical technique that has been proposed as a potential approach for cancer therapy. Meanwhile, berberine (BER), an active ingredient extracted from various medical herbs, such as Coptischinesis, has been proven antitumor effects in a broad spectrum of cancer cells. In this study, we seek to develop a novel dual cancer therapeutic method by integrating pre-administration of BER and LTP exposure and evaluating its comprehensive antitumor effect on the human non-small-cell lung cancer (NSCLC) cell lines (A549 and H1299) in vitro. Cell viability, cell cycle, cell apoptosis, and intracellular and extracellular ROS were investigated. The results showed that cotreatment of BER and LTP significantly decreased the cell viability, arrested the cell cycle in the S phase, promoted cell apoptosis, and increased intracellular and extracellular ROS. Additionally, RNA Sequencing (RNA-Seq) technology was used to explore potential mechanisms. The differentially expressed genes among different treatment groups of NSCLC cells were analyzed and were mainly enriched in the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) signaling pathway. Moreover, cotreatment of BER and LTP notably depressed the total protein expression level of PI3K and AKT with immunoblotting. In conclusion, BER and LTP have a synergistic inhibitory effect on NSCLC cells via the PI3K-AKT signaling pathway, which could provide a promising strategy for supplementary therapy in the anti-NSCLC battle.
Lung cancer is the most common type of malignant tumor worldwide. Plasma-activated medium (PAM) is an innovative cancer treatment method that has received considerable scientific attention. The objective of this study is to evaluate the effects of PAM on the anti-tumor characteristics of non-small cell lung cancer (NSCLC) cells in two-dimensional (2D) and three-dimensional (3D) cultures. The effects of PAM treatment on the proliferative and migratory capabilities of A549 cells in 2D and 3D cultures were assessed using MTT, migration, invasion assays, and cell cycle, respectively. The study also investigated the impact of PAM treatment on the changes in the content of intracellular and extracellular reactive species and analyzed protein expression using the Western Blot method. PAM treatment inhibited the viability, migration, and invasion abilities of A549 cells in both 2D and 3D cultures, suppressed the epithelial–mesenchymal transition (EMT) process, and downregulated the expression of the RAS/ERK signaling pathway, which effectively inhibited tumor spheroid formation. Additionally, the effect of PAM on A549 cells was mediated through ROS-induced oxidative reactions, and PAM treatment exhibited greater cytotoxicity in 2D culture compared to 3D culture. As compared to 2D, the 3D cell culture model provides a viable in vitro cell model for studying the mechanisms of PAM treatment in lung cancer. PAM represents an effective new treatment for NSCLC.
The effective growth and maturation of porcine oocytes in vitro are critical for advancing reproductive biotechnologies. In this study, we explored low-temperature plasma (LTP) treatment as a redox modulation strategy to enhance the survival and maturation of denuded porcine oocytes during in vitro primary culture in order to improve animal cellular health through innovative interventions. Freshly isolated oocytes were exposed to plasma treatment for different lengths of time and subsequently cultured under established in vitro conditions. Morphological and redox-related analyses showed that LTP treatment was associated with increased oocyte diameter, a higher first polar body extrusion rate, mitochondrial membrane potential changes, and altered intracellular and extracellular redox-related parameters. These beneficial effects exhibited a distinct time-dependent dose–response pattern. Furthermore, Western blot analysis showed altered expression of the EGFR/ERK signaling cascade and proteins such as Nrf2, suggesting that LTP treatment might participate in the regulation of maturation-related responses in porcine oocytes cultured in vitro by inducing redox-associated changes, along with alterations in EGFR/ERK-related signaling, Nrf2 expression, and molecules involved in maturation and apoptosis. Collectively, these findings highlight the positive role of LTP in supporting porcine oocyte maturation during in vitro primary culture and provide a promising approach for optimizing the in vitro primary culture of porcine oocytes.