Journal of Cell and Molecular Research

Abstract TGF-β group member myostatin (MSTN) inhibits the growth and differentiation of skeletal muscle. It is unknown how total inhibition of MSTN restricts the hyperproliferation of muscle cells, though. This study aimed to determine the differentially expressed genes and biological pathways associated with myostatin in mouse C2C12 myogenic cells and skeletal muscle tissue, utilizing high-throughput mRNA expression data. In both tissue and cell line, two experimental groups were compared, including wild-type vs. MSTN-Knockout. Transcriptome data were extracted from GEO and analyzed using the LIMMA package in the R environment and GEO2R. Significantly differentially expressed genes were considered as adjusted p-value < 0.05 with log fold change > |0.5|. Lastly, the possible biological pathways were examined with the KEGG database, and the protein-protein interaction (PPI) network was created using Cytoscape software. To find key genes in the PPI network, a topological analysis was conducted using the Network Analyzer tool in Cytoscape software. The total number of differentially expressed genes was 14549 for the C2C12 cell line and 45267 for mouse skeletal muscle tissue; among them, 235 and 1425 transcripts were significant with an adjusted p-value <0.05. The comparison between these two DEG lists showed that 49 genes were common between the myogenic C2C12 cell and skeletal muscle tissue. Additionally, three biological pathways between the C2C12 cell line and the skeletal muscle tissue were in common, and they were associated with the decrease of MSTN gene expression, including: hypertrophic cardiomyopathy, Axon guidance, and dilated cardiomyopathy. These pathways were all related to muscle tissue. Despite the large number of DEGs, only 49 were in common in tissue and cell line; this could indicate that comparing a tissue with its relevant cell line at the transcriptome level might not be precise enough to draw a solid conclusion. This is due to the nature of cell heterogeneity of most tissues, including skeletal muscle.

Abstract Myeloid Zinc Finger 1 (MZF1) is a member of the SCAN domain-zinc finger (SCAN-ZFP) transcription factor family. With a structure comprising five distinct domains, it regulates various transcriptional processes. Numerous studies suggest that MZF1 has bi-functional roles, acting either as an oncogene or a tumor suppressor. However, its overexpression has also been associated with negligible changes in cellular functions, an aspect that remains underexplored. Additionally, although the HEK293T cell line is widely used to investigate molecular mechanisms, the role of MZF1 in these cells has not been reported. In this study, we assessed the effects of MZF1 overexpression on some cellular functions in these cells. Using transposon-mediated gene transfer, we generated stably expressing cell lines and assessed their cellular functions, including cell proliferation, cell viability, and cell stemness.  Our findings show that following stable expression of MZF1, the number of cells compared to controls did not reveal a significant difference during 6 time-points over 72 hours. Also, the MTT read absorbance, and as a consequence, cell viability did not change in the MZF1 overexpressing cell line compared to control cells (0.49 against 0.48). By performing the colony formation assay, we did not find any significant changes in the number of colonies or the number of cells in each colony. Also, our soft agar assay did not reveal statistically significant changes in the size and number of colonies between the two groups of MZF1-overexpressing cells and control cells. Taken together, our data provide evidence that the stable overexpression of MZF1 does not alter cellular functions of proliferation, viability, and stemness in HEK293T cells.

Abstract Treatment of cartilage injuries remains a crucial clinical challenge due to the intrinsically limited capacity of this tissue for regeneration. Common treatment approaches are often unable to restore the natural structure and the long-term function of articular cartilage. Therefore, tissue engineering emerges as a promising approach that enhances cartilage regeneration using cell integration, scaffolds, and bioactive molecules. Chitosan stands out among various types of bioactive materials owing to its biocompatibility, biodegradability, anti-inflammatory effects, and structural similarity to cartilage extracellular matrix, glycosaminoglycans. This review investigates recent developments in the design and application of chitosan scaffolds in cartilage tissue engineering. Various scaffold formats, including hydrogels, porous and nanofibrous structures, as well as three‑dimensional printed constructs, have been shown to support the proliferation, adhesion, and differentiation of chondrocytes. On the other hand, chitosan carriers have been developed for the controlled release of growth factors, anti-inflammatory drugs, and nucleic acids, enhancing the tissue regeneration results. Engineering strategies such as chemical modification, combination with natural or synthetic polymers, and incorporation of bioactive molecules lead to improved mechanical strength, bioactivity, and immunomodulatory properties of chitosan-based scaffolds. Preclinical studies in animal models followed by initial clinical trials and subsequent production of commercial products present promising evidence of the clinical translatability of chitosan scaffolds. Recent innovations, including responsive smart scaffolds and 4D bioprinting, show that scaffolds are transitioning from static biomaterials toward dynamic structures with compatibility in physiologic environments. Overall, chitosan provides a multipurpose and promising biomaterial for cartilage regeneration. The continuation of interdisciplinary research, along with advancements in customized modeling, can accelerate the development of new-generation scaffolds, ultimately leading to improved long-term clinical outcomes.
 

Abstract Breast cancer is the second most common cancer worldwide and the most prevalent cancer among women, causing a large number of deaths annually. This fact underscores the importance of studying the risk factors, diagnostic methods, and treatments for this disease. Single-nucleotide polymorphisms are the most common type of genetic variation in eukaryotic genomes and are found in many genes associated with various cancers. Depending on their location within different regions of a gene, these genetic variants can differently affect gene expression and cancer susceptibility. In this case-control study, we investigated the single-nucleotide polymorphisms of the GPR30/GPER-1 gene, including rs3808350, rs3808351, and rs11544331, and their association with breast cancer risk. The study was conducted on 70 breast cancer patients and 70 healthy women from the female population of Markazi Province, Iran. Blood samples were collected from all participants, and DNA was extracted. The target polymorphisms were genotyped using the tetra-primer amplification refractory mutation system PCR (tetra-ARMS PCR) method. Data analysis was performed using SPSS Statistics 26 and SNP Analyzer 2 software. The results showed a significant association between the AG genotype (co-dominant model) and the combined GG and AG genotypes (dominant model) of rs3808350, indicating an increased risk of breast cancer. For rs11544331, the TT genotype (recessive model) and the combined TT and CT genotypes (dominant model) were also significantly associated with higher risk. No significant association was observed for rs3808351. In conclusion, rs3808350 and rs11544331 polymorphisms may serve as potential biomarkers for early detection and risk assessment of breast cancer. Identifying such genetic markers could enhance diagnostic accuracy and support the development of personalized prevention and treatment strategies.

Abstract Helicobacter pylori is a Gram-negative bacterium that persistently colonizes the gastric mucosa, leading to various gastrointestinal disorders, including gastritis, peptic ulcers, and gastric cancer. Chronic H. pylori infection triggers inflammatory responses and modulates host signaling pathways, particularly receptor tyrosine kinases (RTKs), which regulate essential cellular processes such as adhesion, migration, proliferation, and survival. Among these, EphA2 is a key RTK involved in epithelial cell signaling and maintaining structural integrity, while E-cadherin plays a critical role in cell-cell adhesion and tumor suppression. Disruptions in these signaling molecules can contribute to gastric epithelial dysfunction and cancer progression. This study aimed to evaluate the effects of an Iranian H. pylori vacA d1/-i1 strain on the expression of EPHA2 and E-cadherin in AGS gastric epithelial cells. Following co-culture with H. pylori, total RNA was extracted, and quantitative real-time PCR (qRT-PCR) was performed to analyze gene expression levels. Hematoxylin and eosin (H&E) staining was used to assess morphological changes, revealing structural alterations indicative of cellular stress and epithelial disruption after H. pylori infection. Gene expression analysis demonstrated a significant downregulation of EPHA2 in H. pylori-infected cells compared to controls (p < 0.0001), suggesting bacterial interference with EPHA2-mediated signaling pathways. In contrast, no significant change in E-cadherin expression was observed, indicating that H. pylori may not strongly impact cell adhesion under these conditions. The selective suppression of EPHA2, along with observed morphological changes, suggests a novel mechanism by which H. pylori contribute to gastric epithelial dysfunction, potentially promoting tumorigenesis. Further research is needed to elucidate the molecular pathways involved and their implications for gastric disease pathogenesis and targeted therapeutic strategies.

Abstract Autophagy can play an inhibitory role in the early stages of cancer, so studying autophagy genes can aid in developing therapeutic strategies.  This study investigated the effect of lactoferrin (Lf) and nano-encapsulated Lf (NLf) on the expression levels of autophagy-inducing genes Beclin 1, p53 and LC3I. We also examined the effect of Lf and NLf on the growth rate of breast cancer cell and normal cell lines (MCF7 and Vero). The results showed that the survival percentage of MCF7 cells in NLf treatment was lower than Lf with the same concentrations. The survival of Vero cells treated with NLf and Lf was higher than that of MCF-7 cancer cells, and cell death in this normal cell line was minimal.The level of the Beclin 1gene expression in concentrations of 200 and 300 µg of NLf and Lf increased significantly compared to the control in cancer cells. The expression levels of p53 and LC3I genes were significantly increased in all concentrations of NLf and Lf compared to the control in cancer cells.  The expression levels of all three genes in different concentrations of NLf in normal cells showed a significant increase. The level of gene expression and cell death by NLf was slightly higher than that of Lf in cancer cells, which suggests that the intracellular penetration of NLf was greater than Lf. Lf causes cell survival and protects cells during stress by inducing autophagy in normal cells and removing damaged organelles, and causes cell death in cancer cells in the early stages.