Journal of cellular biochemistry最新文献

The RANK/RANKL/OPG signaling pathway plays a crucial role in breast cancer progression and metastasis. However, its expression patterns and potential implications in breast cancer stem cells remain poorly understood. This study aimed to characterize the expression profile of this pathway in breast cancer stem cells isolated from two distinct breast cancer cell lines: MDA-MB-231 and MCF-7. Mammospheres (MS), representing breast cancer stem cells, were generated using agar-coated 6 well tissue culture plates in suitable mammospheres culture conditions. Flow cytometric analysis showed enrichment of the CD44⁺/CD24⁻ subpopulations in the mammospheres cultures, with MDA-MB-231 exhibiting a higher percentage compared to MCF-7. The isolated MS from both cell lines showed upregulation of stemness markers OCT4 and SOX2, with MS. MDA-MB-231 demonstrating higher expression levels. Analysis of the RANK/RANKL/OPG axis revealed differential expression patterns between the two cell lines. RANK expression was significantly upregulated in MS. MDA-MB-231 but not in MS. MCF-7. Interestingly, while OPG mRNA levels were elevated in mammospheres from both cell lines, secreted OPG protein levels were paradoxically reduced in the mammospheres conditioned media. Additionally, RUNX2, an osteoblastic marker, and a downstream target of RANK signaling, showed a decreased expression in both mammospheres compared to adherent cells. These findings suggest a complex, context-dependent regulation of the RANK/RANKL/OPG pathway in breast cancer stem cells, potentially contributing to the aggressive nature and metastatic propensity of triple-negative breast cancer. This study provides novel insights into the molecular characteristics of breast cancer stem cells and underscores the complexity of OPG/RANK/RANKL axis expression in them; a role yet to be fully elucidated.

This study endeavors to assess the potential protective role of bergapten (BP) in mitigating abdominal aortic aneurysm (AAA) and to decipher the underlying mechanisms and molecular targets. Network pharmacology was utilized to search for potential targets of BP against AAA. Molecular docking and molecular dynamics simulations were utilized to validate the interaction of BP with core targets, and then the therapeutic effect and mechanism of BP on AAA were verified by using an elastase-induced AAA model. Network pharmacology analysis identified five pharmacological targets for BP, including EGFR, SRC, PIK3CA, PIK3CB, and JAK2. Molecular docking and molecular dynamics simulations further prioritized JAK2 as the most promising candidate for the potential treatment of AAA. The results of animal experiments demonstrated that BP significantly reduced the expression of inflammatory cytokines IL-6, TNF-α, and IL-1β in the aortic tissue of AAA mouse model, and inhibited the phosphorylation of JAK2 and STAT3. BP plays an important role in the treatment of AAA, and it may become a promising drug to combat AAA progression. The inhibitory effect of BP on AAA vascular progression and the attenuation of inflammatory cell infiltration may be related to the regulation of JAK2/STAT3 signaling pathway.

Aurora kinases (AKs), a family of serine/threonine kinases, play a vital role in chromosome segregation during the cell cycle (Mountzios et al., 2008). This family includes Aurora Kinase A (AKA), Aurora Kinase B (AKB), and Aurora Kinase C (AKC). AKA and AKB are active during mitosis, while AKC is involved mostly in germ cell as well as somatic cells. Elevated levels of AKC have been found in several cancer cell lines including breast, cervical, thyroid, colorectal, and liver cancers, making it a significant target for cancer therapy (Tang et al., 2017). In cancers such as glioblastoma and prostate cancer, for example, AKC up regulation has been associated with increased tumor aggressiveness, highlighting its potential role in tumor progression and poor prognosis. Our study employs computational methods, including molecular docking and structure-based virtual screening, to explore a data set of 2 65 241 compounds from the National Cancer Institute (NCI) database, focusing on AKC as a potential target for drug discovery. Through docking studies, several promising compounds that interact with the enzyme's ATP binding pocket, particularly with residues Phe54, Lys72, Ala123, Glu121 and Glu127 of AKC, were identified. The stability of these interactions was assessed through 200-ns molecular dynamics (MD) simulations, revealing that the majority of compounds exhibited stable interactions, while a few displayed fluctuations in their trajectories. Most compounds adhered to favorable pharmacokinetic properties. Comprehensive MD simulations and free energy calculations identified three top candidates (90 729, 37 623, and 134 546) with strong potential as potent inhibitors of AKC. Additional in vitro and in vivo studies are required to confirm the therapeutic potential of these candidates.

Claudin-1 (CLDN1) and CLDN4 are highly expressed in keratinocytes and may function as a paracellular barrier to water and small molecules. The physiological function of CLDN4 has not been fully understood, whereas dysfunction of CLDN1 is involved in the pathophysiology of allergy and inflammatory diseases. Here, we found that the protein level of CLDN4 in the skin tissues of 36-week-old mice was lower than that in 6-week-old mice. In contrast, there was not much difference in the mRNA levels of CLDN4. Tenovin-1 (Ten-1), a sirtuin-1/2 inhibitor, decreased the protein level of CLDN4 without affecting that of CLDN1 in human keratinocyte-derived HaCaT cells. The decrease in CLDN4 mRNA by Ten-1 was much less than that in protein. Cycloheximide-chase assay showed that the protein stability of CLDN4 was attenuated by Ten-1. The Ten-1-induced decrease in CLDN4 protein was inhibited by clathrin-dependent endocytosis and proteasome inhibitors. The Ten-1 treatment or SIRT2 silencing induced the elevation of acetylated CLDN4 protein, leading to the reduction of CLDN4 protein. In addition, the paracellular barrier function was reduced by Ten-1 treatment or SIRT2 silencing. These results indicate that Ten-1 may enhance the clathrin-dependent endocytosis and proteasome-dependent degradation of CLDN4 protein, resulting in the dysfunction of paracellular barrier. The Ten-1-induced reduction of CLDN4 protein and paracellular barrier function were inhibited by curcumin, a polyphenol contained in Curcuma longa plant. We suggest that the reduction of CLDN4 protein in keratinocytes may be involved in the age-related dysfunction of the skin barrier, which may be rescued by curcumin.

PANoptosis is involved in various pathological processes, but its role in acute myocardial infarction (AMI) remains unclear. This study aimed to explore the mechanism by which PANoptosis is involved in AMI. GSE172270 was used as an internal test set, and GSE159657 served as an external validation set to identify disease targets for AMI. WGCNA was performed to identify potential hub genes associated with AMI. Then, genes related to PANoptosis among the disease targets of AMI were screened. Additionally, human cardiomyocytes AC16 and mouse cardiomyocytes HL-1 were cultured in sugar-free and serum-free medium for 4 h and 12 h, and the expression of key genes regulating PANoptosis was detected by qRT-PCR. Finally, mRNA interference and overexpression experiments were conducted to verify the expression of key mRNAs. A total of 45 upregulated and 125 downregulated differentially expressed genes (DEGs) were identified in the GSE172270 data set. WGCNA identified 891 potential hub genes associated with AMI. In the GSE159657 data set, 695 upregulated and 552 downregulated DEGs were identified. Four genes related to AMI-PANoptosis (PRDX1, MMACHC, BLVRB, and TXNL1) were screened. Through qRT-PCR verification, PRDX1 was identified as the most specific and significant gene. The expression of the PANoptosis positive regulator ZPB1 was upregulated, while the PANoptosis negative regulator TAK1 was downregulated. Additionally, key cell death genes (MLKL, p-MLKL, Caspase-3, Caspase-7, and GSDMD) were upregulated in sugar-free and serum-free culture. However, PRDX1 interference reversed these effects. Our study demonstrated that PRDX1 is a key regulator of PANoptosis in AMI.

Breast cancer is one of the most common types of cancer in women, and metastasis is a leading cause of mortality in patients with this disease. This study investigated the effects of melatonin, a natural hormone, on the migration of cancer cells in two cell lines, MCF-7 and MDA-MB-231. MCF-7 and MDA-MB-231 cells were cultured in their respective media. The effective dose of melatonin in each cell line was determined using the MTT assay. The effects of IC50 melatonin on cell migration were assessed using the wound-healing assay. The expression of the invasion-related genes (MMP2 and MMP9), as well as the melatonin receptors MT1 and MT2, was analyzed using Real-Time RT-PCR. The wound-healing assay results indicated that 48 h of melatonin treatment at doses of 2.5 and 3.5 M significantly reduced migration in MCF-7 and MDA-MB-231 cells. In addition, melatonin treatment decreased the invasion-related markers of both cell lines. Melatonin also increased the expression of MT1 and MT2 receptors in both cell lines, and the expression of MMP2 and MMP9 was significantly reduced by melatonin (p < 0.05). Our results indicate that melatonin, a naturally occurring compound, possesses the potential to inhibit the movement and spread of breast cancer cells by elevating the levels of MT1 and MT2 receptors, resulting in a reduction of matrix metalloproteinases 2 and 9 expression.

Glomerular filtration function and homeostasis are largely due to the cross-talk between podocytes, endothelial cells, and mesangial cells (MCs). Any disturbance in this association causes glomerular diseases (GD). Cell-based therapies are the best option in the treatment of GD. It is contemplated that hematopoietic stem cells (HSCs) are best suited to regenerate these cells; earlier, we have shown the differentiation of HSCs into podocytes. In this study, MCs formation was initiated with retinoic acid (RA), BMP-7, and Activin A, resulting in comma-shaped intermediate mesoderm (IM) cells prominently expressing Osr1. Followed by inducing with EGF, FGF, and BMP-7, which resulted in elongated metanephric mesenchyme (MM) cells conspicuously expressing Pax2, Wt1, Foxd1, and Eya1. Finally, MM cells were induced with platelet-derived growth factor to form polygonal-shaped MCs expressing α-smooth muscle actin, desmin, CD44, and PDGFRβ. The growing MCs showed positivity to periodic acid Schiff's, and ANAE staining with a prominent expression of the Itga8 elucidating phagocytic property of MCs. These MCs showed conspicuous expression of CD133, notch-2, and telomerase, determining the quiescence nature with a 31.2% proliferation rate revealed through Ki-67 staining. The functionality of MCs was assessed by growing MCs in 5.5 and 25 mM glucose concentrations, and noticeable expression of angiotensinogen, angiotensin-I and II, angiotensin-converting enzyme, collagen-4, fibronectin, and TGFβ1 was observed in 25 mM concentration, while lowered expression of these genes was observed in 5.5 mM concentration explaining the role of MCs in regulating the filtration pressure. All these findings demonstrate that HSCs can rejuvenate the insulted glomerulus.

The insulin-like growth factor 1 receptor (IGF1R) is a crucial receptor tyrosine kinase involved in cellular growth, survival, and metabolism. Abnormal overexpression and activation are common in various cancers and contribute to tumor development and resistance to treatment. The STRING database was used to analyze the protein–protein interaction network of IGF1R and was visualized using Cytoscape to identify the key associated proteins. We assessed IGF1R and its associated protein expression levels across pan-cancer types and compared them to healthy controls using a TNMplot and cBioPortal. The objective of this study was to identify novel, low-toxicity inhibitors targeting the IGF1R and its associated proteins (e.g., AKT1 and EGFR) with better pharmacokinetic profiles for effective cancer treatment, including brain cancer. We screened 693 million drug-like compounds and selected the top 400 for toxicity analysis using ProTox-II, which identified 83 nontoxic candidates. These were categorized as either blood–brain barrier (BBB) permeant or impermeant. Molecular docking studies with AutoDock Vina 4.1 were performed on 17 target proteins, including IGF1R, with the top three compounds. Subsequently, molecular dynamics simulations using Desmond were conducted on the two most promising candidates: two BBB permeants and two impermeants. Our study identified six nontoxic IGF1R inhibitors and 16 other target protein inhibitors. Docking and MD simulations confirmed the potential of these compounds in targeted therapies. Notably, both BBB-permeant and -impermeant compounds in complex with the target proteins showed stability over 50 and 400 ns molecular simulation experiments, highlighting their potential in cancer therapy and suggesting the need for further in vitro and in vivo validation.