Proliferation of SH-SY5Y neuroblastoma cells on confined spaces.

BACKGROUND: Microfluidics offers precise drug delivery and continuous monitoring of cell functions, which is crucial for studying the effects of toxins and drugs. Ensuring proper cell growth in these space-constrained systems is essential for obtaining consistent results comparable to standard Petri dishes. NEW METHOD: We investigated the proliferation of SH-SY5Y cells on circular polycarbonate chambers with varying surface areas. SH-SY5Y cells were chosen for their relevance in neurodegenerative disease research. RESULTS: Our study demonstrates a correlation between the chamber surface area and SH-SY5Y cell growth rates. Cells cultured in chambers larger than 10 mm in diameter exhibited growth comparable to standard 60-mm dishes. In contrast, smaller chambers significantly impeded growth, even at identical seeding densities. Similar patterns were observed for HeLaGFP cells, while 16HBE14σ cells proliferated efficiently regardless of chamber size. Additionally, SH-SY5Y cells were studied in a 12-mm diameter sealed chamber to assess growth under restricted gas exchange conditions. COMPARISON WITH EXISTING METHODS: Our findings underscore the limitations of small chamber sizes in microfluidic systems for SH-SY5Y cells, an issue not typically addressed by conventional methods. CONCLUSIONS: SH-SY5Y cell growth is highly sensitive to spatial constraints, with markedly reduced proliferation in chambers smaller than 10 mm. This highlights the need to carefully consider chamber size in microfluidic experiments to achieve cell growth rates comparable to standard culture dishes. The study also shows that while SH-SY5Y and HeLaGFP cells are affected by chamber size, 16HBE14σ cells are not. These insights are vital for designing effective microfluidic systems for bioengineering research.

Deciphering the Role of miR-30a-5p and DLGAP1 Gene in Non-small Cell Lung Cancer.

BACKGROUND/AIM: Non-small cell lung cancer (NSCLC) is the deadliest form of cancer worldwide. Understanding the mechanisms of lung cancer development is vital for targeted therapy advancements. This article explores the little-known role of the guanylate kinase-associated protein (GKAP), encoded by the Disks large-associated protein 1 (DLGAP1) gene, in NSCLC along with assessing microRNA-30a-5p's influence on DLGAP1 gene expression in the A549 cell line. MATERIALS AND METHODS: Experiments were conducted on A549 cells transfected with synthetic oligonucleotides. The luciferase assay was employed to confirm the binding site of miR-30a-5p to the 3'UTR of DLGAP1 mRNA. The role of miRNA-30a-5p mimic in regulating potential target gene expression at the protein and mRNA levels was studied by performing RT-qPCR and western blot analyses. The effects of DLGAP1 knockdown and miRNA-30a-5p mimic on cell viability and the cell cycle were evaluated using the MTT test and flow cytometry with annexin/iodide cell staining. RESULTS: The luciferase assay indicated that miR-30a-5p has the ability to bind to the 3'UTR of DLGAP1 mRNA. RT-qPCR revealed that the overexpression of miR-30a-5p down-regulates DLGAP1 mRNA. Western blot analysis indicated that miR-30a-5p slightly reduces the level of the GKAP protein. Knockdown of DLGAP1 with synthetic oligonucleotides, as well as transfection with a miR-30a-5p mimic, significantly attenuates cell proliferation and increases the number of cells in the early and late stages of apoptosis. CONCLUSION: Our findings reveal the antiproliferative effect of miR-30a-5p and DLGAP1 gene knockdown on A549 cancer cells, implying that these elements could be considered as therapeutic targets for personalized medicine in NSCLC patients.

Mitochondrial Dynamics in Neurodegenerative Diseases: Unraveling the Role of Fusion and Fission Processes.

Neurodegenerative diseases (NDs) are a diverse group of disorders characterized by the progressive degeneration and death of neurons, leading to a range of neurological symptoms. Despite the heterogeneity of these conditions, a common denominator is the implication of mitochondrial dysfunction in their pathogenesis. Mitochondria play a crucial role in creating biomolecules, providing energy through adenosine triphosphate (ATP) generated by oxidative phosphorylation (OXPHOS), and producing reactive oxygen species (ROS). When they're not functioning correctly, becoming fragmented and losing their membrane potential, they contribute to these diseases. In this review, we explore how mitochondria fuse and undergo fission, especially in the context of NDs. We discuss the genetic and protein mutations linked to these diseases and how they impact mitochondrial dynamics. We also look at the key regulatory proteins in fusion (MFN1, MFN2, and OPA1) and fission (DRP1 and FIS1), including their post-translational modifications. Furthermore, we highlight potential drugs that can influence mitochondrial dynamics. By unpacking these complex processes, we aim to direct research towards treatments that can improve life quality for people with these challenging conditions.

Role of the glucosinolates in cancer epigenetics / Rola glukozynolanów w epigenetyce nowotworów.

It has been known for years that diet impacts human health, including the risk of cancer development. Food components can both increase and reduce the risk of carcinogenesis. Thereby, a wisely composed diet can extend life span and improve life quality. The favourable effect on health exert glucosinolates (GSLs), a group of secondary plant metabolites found in vegetables of the Brassicaceae family, such as broccoli, cauliflower, cabbage, and kohlrabi. Hydrolysis of GSLs results in the formation of compounds, like sulforaphane (SFN), phenylethyl isothiocyanate (PEITC) and 3,3'-Diindolylmethane (DIM), which are known for versatile anti-cancer activity. This review highlights advances on the role of the chosen GSLs on selected epigenetic mechanisms, i.e. DNA methylation, histone acetylation and microRNAs expression in cancer treatment.

The Role of Bcl-xL Protein in Viral Infections.

Bcl-xL represents a family of proteins responsible for the regulation of the intrinsic apoptosis pathway. Due to its anti-apoptotic activity, Bcl-xL co-determines the viability of various virally infected cells. Their survival may determine the effectiveness of viral replication and spread, dynamics of systemic infection, and viral pathogenesis. In this paper, we have reviewed the role of Bcl-xL in the context of host infection by eight different RNA and DNA viruses: hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus (HIV), influenza A virus (IAV), Epstein-Barr virus (EBV), human T-lymphotropic virus type-1 (HTLV-1), Maraba virus (MRBV), Schmallenberg virus (SBV) and coronavirus (CoV). We have described an influence of viral infection on the intracellular level of Bcl-xL and discussed the impact of Bcl-xL-dependent cell survival control on infection-accompanying pathogenic events such as tissue damage or oncogenesis. We have also presented anti-viral treatment strategies based on the pharmacological regulation of Bcl-xL expression or activity.

Serum miR-210-3p as a Potential Noninvasive Biomarker of Lung Adenocarcinoma: A Preliminary Study.

Background: Development of noninvasive biomarkers could potentially contribute to extending the 5-year overall survival rate of nonsmall cell lung cancer (NSCLC) patients. Circulating microRNAs (miRNAs), due to their high stability, have the potential to become valuable cancer biomarkers. Methods: Using reverse transcription-quantitative polymerase chain reaction and testing three methods for data normalization, the expression levels of six miRNAs were evaluated in serum samples obtained from 50 NSCLC patients. Subsequently, the clinical significance of the tested miRNAs was determined. Results: Significant downregulation of miR-21-5p, miR-30a-5p, and miR-126-3p and upregulation of miR-210-3p and miR-486-5p in serum samples of NSCLC patients were identified in comparison to healthy controls. miR-205-5p appeared to be undetectable in all tested samples. Furthermore, miR-210-3p was differentially expressed between two subtypes of NSCLC. Receiver operating characteristic analysis for miR-210-3p revealed the area under the curve of 0.842 (95% confidence interval, 0.72-0.96; p = 0.0003) and demonstrated that miR-210-3p displayed considerable accuracy in discriminating between lung adenocarcinoma (AC) patients and healthy controls. Conclusions: Findings from this preliminary study indicate that five of six tested miRNAs were deregulated in the serum of NSCLC patients. Moreover, miR-210-3p appears to be a promising biomarker for diagnosis of lung AC.

Forms of diagnostic material as sources of miRNA biomarkers in hepatocellular carcinoma: a preliminary study.

Aim: To assess the diagnostic value of selected miRNAs from various material collected from hepatocellular carcinoma (HCC) patients. Patients & methods: Tissue, serum, urine and fecal samples from HCC patients and healthy individuals were screened for associated miRNAs using microarray analysis; the selected miRNAs were then validated by real time-quantitative PCR on 65 patients. Results: Serum miR-122, a combination of serum miR-155 with miR-885-5p, a combination of urinary miR-532-3p with miR-765, and fecal miR-320a displayed 100% efficiency in discriminating patients from controls. A combination of urinary miR-532-3p and miR-765 allowed patients with neoplastic grade G3 to be distinguished from those with G1 and G2. Conclusion: Additionally to serum, urine and feces also appeared to be valuable source of potential HCC noninvasive miRNA biomarkers.

miR-30a-5p together with miR-210-3p as a promising biomarker for non-small cell lung cancer: A preliminary study.

BACKGROUND: Although an immense effort has been made to develop novel diagnostic methods and treatment strategies for non-small cell lung cancer (NSCLC), the survival rate of this disease has remained virtually unchanged. Small non-coding RNAs called microRNAs (miRNAs) have appeared to be very promising biomarkers of cancer including NSCLC. OBJECTIVE: We investigated the expression level of six miRNAs, and subsequently we evaluated their diagnostic ability and their clinical significance. METHODS: We performed an analysis in 50 paired cancer and non-cancerous lung tissue samples collected from NSCLC patients. The RT-qPCR technique was used to investigate the expression profile. RESULTS: Obtained results indicate that miR-30a-5p, miR-126-3p and miR-486-5p are downregulated, while miR-205-5p and miR-210-3p are upregulated in NSCLC tissue. Moreover, performed stepwise discriminant analysis determined the model including miR-30a-5p and miR-210-3p which tested on the test set (n= 30) revealed an AUC of 0.969 and provided 100% sensitivity and 80% specificity in discriminating NSCLC tissue from non-cancerous lung tissue. CONCLUSIONS: The present preliminary study demonstrated that five tested miRNAs were deregulated in cancer tissue. Moreover, miR-30a-5p together with miR-210-3p with excellent sensitivity and acceptable specificity may distinguish cancer tissue form non-cancerous tissue and thus may become a potential diagnostic biomarker for NSCLC.

Circulating miRNAs as non-invasive biomarkers for non-small cell lung cancer diagnosis, prognosis and prediction of treatment response.

The discovery of a new class of molecules, the 22-24 nucleotides long RNA, has initiated a new chapter in genetic information regulation studies. These molecules have a significant impact on the functioning of cells by means of negative regulation of expression of targeted mRNA. Special attention is given to exogenous, circulating miRNAs, whose levels of expression in different body fluids varies and can be deregulated by pathological, as well as physiological conditions. Extensive studies on the diagnostic potential of miRNAs are currently conducted. Attempts are made to determine specific profiles of miRNAs that will correlate with disease entity. The preliminary data gives hope that in the near future miRNAs will be applied as a diagnostic and prognostic biomarkers in many diseases. One of their most significant applications can be in the diagnosis of lung cancer - the most deadly form of cancer, due to its too late diagnosis. Abundant in miRNA molecules, blood and sputum constitute excellent diagnostic material for patients with lung cancer, especially those with non-small-cell lung cancer NSCLC. Easy procedures for obtaining this diagnostic material from patients and relatively easy analysis of miRNA expression, make these molecules promising in their use in diagnosis, predicting of therapy effects and prognosis for patients with NSCLC. This work presents information related to miRNA and their specific attributes. Moreover level of deregulated circulating miRNAs occurring in body fluids of patients with NSCLC is also discussed.

Treatment with platelet-derived growth factor (PDGF) and rock inhibitors is related to declined nerve growth factor (NGF) signaling in an experimental model of rat pulmonary hypertension.

BACKGROUND: Recent studies reveal that nerve growth factor (NGF) plays a critical role in the pathobiology of pulmonary hypertension (PH). The aim of the present study is to clarify the relationship between NGF signaling and treatment with PDGF or ROCK inhibitors in an animal model of PH. METHODS: Lung tissues were obtained from animals with monocrotaline (MCT)-challenged PH which had been administered long term imatinib, fasudil or statin. Reversal of disease was indicated by decreases in right ventricle pressure (RVP) and hypertrophy. NGF expression was examined at the mRNA and protein levels using quantitative real-time PCR reaction and ELISA. RESULTS: MCT significantly increased NGF mRNA and protein content in lung tissue. ROCK inhibitor (fasudil) and PDGF inhibitor (imatinib) caused significant decreases in NGF mRNA and protein content when administered alone, with no further effects noted when used in combination. CONCLUSION: The beneficial reversal of MCT-mediated effects in PH caused by PDGF or ROCK inhibition may be also partially mediated by decreased NGF signaling.