Gallbladder cancer (GBC) is a biliary tract cancer with a poor prognosis. Consistent evidence suggests that fasting has extensive antitumor effects in various cancers and influences levels of poly (rC)-binding protein 2...Gallbladder cancer (GBC) is a biliary tract cancer with a poor prognosis. Consistent evidence suggests that fasting has extensive antitumor effects in various cancers and influences levels of poly (rC)-binding protein 2 (PCBP2). However, whether fasting and PCBP2 are involved in GBC remains unknown. We assessed the expression of PCBP2 in GBC tumor tissues and cells. Knockdown and overexpression of PCBP2, combined with in vitro and in vivo assays using fasting mimic medium or diets, were conducted to provide functional significance. The effect of PCBP2 on glycolysis was assessed by glucose uptake, lactate production, oxygen consumption rate, and limiting glycolytic-associated enzymes (PDK1, PKM2, and HK-2). We found that fasting could inhibit glycolysis and cell migration/invasion in GBC cells and that fasting mimic diets could significantly inhibit GBC cell proliferation in a mouse xenograft model. PBCP2 was upregulated in GBC tumor tissues and cells. Moreover, PCBP2 is a key downstream target of fasting, and fasting decreases PCBP2 expression in GBC cells. PCBP2 knockdown inhibits GBC cell proliferation, migration/invasion, and glycolysis, whereas PCBP2 overexpression has the opposite effect. Through co-immunoprecipitation, we identified a physical connection between PCBP2 and the angiopoietin-like protein ANGPTL4. PCBP2 can negatively regulate the expression of ANGPTL4. Hence, fasting inhibits cell proliferation, migration/invasion, and glycolysis through PCBP2/ANGPTL4 signaling. We conclude that PCBP2 is a target of fasting and is involved in cell migration/invasion and glycolysis through the negative regulation of ANGPTL4 in GBC. PCBP2 represents a potential therapeutic target for GBC.
To explore functions and mechanisms in gastric cancer (GC) progression. The mRNA and protein levels of NSUN2 and phosphoglycerate kinase 1 (PGK1) were determined by qRT-PCR and western blot. Cell proliferation, apoptosis...To explore functions and mechanisms in gastric cancer (GC) progression. The mRNA and protein levels of NSUN2 and phosphoglycerate kinase 1 (PGK1) were determined by qRT-PCR and western blot. Cell proliferation, apoptosis, invasion and stemness were examined using CCK8 assay, EdU assay, flow cytometry, transwell assay and sphere formation assay. Cell glycolysis was evaluated by detecting glucose consumption, lactate production and ATP/ADP ratio. The interaction between PGK1 and NSUN2 or YBX1 was evaluated using MeRIP assay or RIP assay. Actinomycin D treatment assay was used to detect the effect of NSUN2 or YBX1 knockdown on PGK1 mRNA stability. The protein levels of p-PI3K/PI3K and p-AKT/AKT were tested by western blot. Animal study was performed to confirm the effect of NSUN2/PGK1 axis on GC tumorigenesis. NSUN2 was confirmed to be upregulated in GC tissues and cells. NSUN2 silencing could repress GC cell growth, invasion, stemness and glycolysis. NSUN2 enhanced PGK1 mRNA stability through promoting its m5C modification, and this modification could be recognized by YBX1. Besides, PGK1 overexpression reversed the inhibitory effect of NSUN2 knockdown on GC cell growth, invasion, stemness and glycolysis. In addition, NSUN2/PGK1 axis increased the activity of PI3K/AKT pathway. Animal study revealed that interference of NSUN2 reduced GC tumorigenesis via inactivating the PGK1/PI3K/AKT pathway. NSUN2/PGK1 axis might play a vital role in GC development.
Colorectal cancer (CRC) remains a significant global health concern, and reliable biomarkers are needed to improve early diagnosis, prognostication, and personalized treatment strategies. This study investigated the expr...Colorectal cancer (CRC) remains a significant global health concern, and reliable biomarkers are needed to improve early diagnosis, prognostication, and personalized treatment strategies. This study investigated the expression of cell surface proteins and serum exosomal miRNAs in CRC patients. Tissue microarrays (TMAs) constructed from primary and metastatic CRC samples were analyzed for five cell surface proteins: EphB1, EphB3, EphA2, cMet, and EphB4. Immunohistochemistry was performed on the TMAs to validate their expression levels. We found that the distribution of expression for all four receptors, except EphA2, was significantly higher ( < 0.01) in CRC samples compared to non-cancerous tissue. High expression of EphB3 was detected in 37% of patient samples, followed by cMet, which was observed in 35%. Exosomes were isolated from the serum of three CRC patients with tumors exhibiting high expression of LGR5 and/or EphB3, four healthy donors and two CRC cell lines. Serum exosomal miRNA analysis identified miR-3168 as significantly upregulated in CRC patients, showing a 3.8-fold increase compared to healthy controls ( < 0.001) and a 2.6-fold increase in CRC cell lines compared to controls ( = 0.02). Ingenuity Pathway Analysis (IPA) suggested that miR-3168 may regulate cMet, EphB3, and EphB4, along with other CRC-associated molecules and pathways. These findings highlight the potential of EphB3 and cMet as biomarkers in CRC, and miR-3168 as a promising minimally-invasive biomarker for monitoring disease progression and therapeutic response. However, further validation in larger cohorts is needed to establish their clinical utility.
Circular RNA (circRNA) is involved in the occurrence of many cancers. Nonetheless, the mechanism of circ_0003998 in non-small cell lung cancer (NSCLC) needs to be studied in depth. Real-time quantitative PCR (RT-qPCR) wa...Circular RNA (circRNA) is involved in the occurrence of many cancers. Nonetheless, the mechanism of circ_0003998 in non-small cell lung cancer (NSCLC) needs to be studied in depth. Real-time quantitative PCR (RT-qPCR) was carried out to check the expression of circ_0003998, microRNA-330-5p (miR-330-5p), chemokine (C-X-C motif) ligand 3 (CXCL3) and methyltransferase-3 (METTL3) in NSCLC tissues and cells. CXCL3, Vimentin and E-cadherin protein levels were measured by western blot. The functions of circ_0003998 in NSCLC cell proliferation, apoptosis, angiogenesis, migration and invasion were tested by clone formation assay, flow cytometry, tube formation assay, wound healing assay, and transwell assay in vitro. The dual-luciferase reporter assay was made to verify the relationship between miR-330-5p and circ_0003998 or CXCL3. Finally, animal experiment was made to further research the function of circ_0003998 on tumor formation in vivo. The interaction between circ_0003998 and METTL3 was analyzed by RNA Immunoprecipitation (RIP) assay, methylated RNA Immunoprecipitation (MeRIP) assay and dual-luciferase reporter assay. In NSCLC tissue and cells, circ_0003998 was markedly overexpressed. Circ_0003998 suppression inhibited NSCLC cell growth, angiogenesis, migration and invasion. Circ_0003998 sponged miR-330-5p, and miR-330-5p inhibitor could reverse the suppression effect of circ_0003998 knockdown on NSCLC cell behaviors. CXCL3 was a downstream target gene of miR-330-5p, and CXCL3 overexpression also reversed the suppressive effect of miR-330-5p on NSCLC cell behaviors. Interference of circ_0003998 reduced NSCLC tumorigenesis by regulating miR-330-5p/CXCL3 axis. Also, METTL3 promoted the expression of circ_0003998 by m6A modification. METTL3-modified circ_0003998 promoted NSCLC cell malignancy through miR-330-5p/CXCL3 axis, suggesting that circ_0003998 might be a new treatment strategy for NSCLC.
Cancer cells frequently undergo stresses like hypoxia, glucose deprivation, and calcium depletion, leading to protein misfolding and accumulation of unfolded proteins in the ER, which trigger ER stress. The unfolded prot...Cancer cells frequently undergo stresses like hypoxia, glucose deprivation, and calcium depletion, leading to protein misfolding and accumulation of unfolded proteins in the ER, which trigger ER stress. The unfolded protein response (UPR) is activated by endoplasmic reticulum (ER) stress to restore protein homeostasis by regulating protein synthesis and degradation. This review explores the multifaceted role of UPR in tumor growth, chemoresistance, and immune evasion in gynecological cancers, particularly ovarian, endometrial, and cervical cancers. UPR-associated genes have been reported to have a potential role as disease biomarkers and therapeutic targets, thus improving early detection and personalized treatment. This review aims to give insights into the role of UPR pathway in gynecological cancers and offers new perspectives for future research and clinical applications.
Globally, gastric cancer (GC) continues to be the primary cause of death due to cancer. This study aimed to investigate the role of in GC and assess the potential synergistic effects of Baicalin and knockdown on GC cel...Globally, gastric cancer (GC) continues to be the primary cause of death due to cancer. This study aimed to investigate the role of in GC and assess the potential synergistic effects of Baicalin and knockdown on GC cells. Differential expression analysis of GC-related datasets was conducted, and a protein-protein interaction (PPI) network was established. Key targets were screened, and prognostic genes were identified using a Least Absolute Shrinkage and Selection Operator (LASSO) Cox regression model. Functional assays assessed the effects of knockdown and Baicalin treatment on GC cell behavior and pyroptosis. was highly expressed in GC cells, and its knockdown reduced cell behavior, inducing G1 arrest and apoptosis. Combined with Baicalin, these effects were enhanced, synergistically inhibiting GC cell behavior. Detection kits showed that knockdown of or baicalin treatment increased lactate dehydrogenase (LDH) release and reactive oxygen species (ROS) levels, while their combination further exacerbated oxidative stress and cell damage. Western blot (WB) analysis revealed that baicalin combined with knockdown synergistically promoted pyroptosis by activating the NLRP3 inflammasome and regulating the NF-κB-NLRP3-Caspase-1 axis. xenograft models demonstrated that knockdown or combined treatment with baicalin significantly inhibited GC progression. These results suggest that knockdown combined with baicalin can inhibit GC progression by regulating cell behavior, cell cycle, pyroptosis, and the NF-κB-NLRP3-Caspase-1 axis in GC cells. This mechanism is expected to become a new target for GC treatment.
Small cell lung cancer (SCLC) accounts for approximately 15% of primary lung carcinomas and has the poorest outcome in all subtypes of lung cancer. The major hurdle for SCLC treatment failure is resistance to platinum-ba...Small cell lung cancer (SCLC) accounts for approximately 15% of primary lung carcinomas and has the poorest outcome in all subtypes of lung cancer. The major hurdle for SCLC treatment failure is resistance to platinum-based chemotherapy. Therefore, an unmet need is to discover new targets that promote SCLC progression and chemoresistance. Based on the signature of ubiquitination-related genes (URGs), differentially expressed genes between cisplatin-resistant and cisplatin-sensitive SCLC cell lines were identified using the Genomics of Drug Sensitivity in Cancer (GDSC) database. The URGs associated with the prognosis were further screened by Cox and LASSO regression analyses, as well as a Kaplan-Meier survival analysis. The E3 ligase NEDD4 was identified to be associated with cisplatin resistance, poor prognosis and tumor metastasis in SCLC. The functional enrichment analysis indicated that the functions and pathways regulated by NEDD4 were enriched in cell proliferation, cell invasion, as well as ubiquitination and PI3K-AKT pathways in SCLC. The knockdown and overexpression of NEDD4 demonstrated that NEDD4 induced the phosphorylation of AKT in SCLC cells. Cell viability, wound healing and transwell invasion assays demonstrated that NEDD4 promoted the proliferation, chemoresistance and invasion of SCLC cells. These results suggest that NEDD4 is a biomarker of a poor prognosis for SCLC, and that it promotes AKT activation, SCLC progression and chemoresistance.
This study aimed to explore key regulatory molecules involved in metabolic alterations clarify the heterogeneity of glioblastoma and develop novel therapeutic strategies. The microarray dataset GSE45117 was retrieved fro...This study aimed to explore key regulatory molecules involved in metabolic alterations clarify the heterogeneity of glioblastoma and develop novel therapeutic strategies. The microarray dataset GSE45117 was retrieved from the Gene Expression Omnibus database to analyze differentially expressed genes (DEGs) glioma stem cell (GSC) populations were enriched via microsphere suspension culture and ALDH+ cell sorting in vitro with the expression of the uridine-cytidine kinase 2 (UCK2) gene compared between stemness and non-stemness populations the UCK2 gene was stably knocked down or overexpressed in GSCs to assess cell invasion migration glucose uptake lactate production and ATP levels. Database analysis revealed high UCK2 expression in cancer stem cells (CSCs) manipulating UCK2 levels affected stemness factors and cell behaviors including proliferation migration invasion and tumor growth UCK2 was more abundant in hypoxic central tumor regions promoting increased glucose uptake and energy production knocking down UCK2 reduced glycolysis and stem cell properties under hypoxia mechanistically UCK2 stabilizes PI3K protein through deubiquitination thereby activating the Akt/HIF-1α pathway. UCK2 plays a pivotal role as a metabolic regulator in glucose metabolism by stabilizing PI3K protein expression via deubiquitination which in turn activates the Akt/HIF-1α signaling pathway.
NRF2, a crucial antioxidant transcription factor in ovarian cancer (OC), is closely associated with CEBPB activation. However, the regulatory mechanism of NRF2 by CEBPB in OC remains poorly understood. In this study, we...NRF2, a crucial antioxidant transcription factor in ovarian cancer (OC), is closely associated with CEBPB activation. However, the regulatory mechanism of NRF2 by CEBPB in OC remains poorly understood. In this study, we systematically evaluated the malignant behavior of SKOV3 and A2780 cells through comprehensive approaches, including CCK-8 kits, clone formation assays, and flow cytometry analysis. Cellular antioxidant capacity was quantitatively assessed using the DCFH-DA and total-antioxidant capacity (T-AOC) assays. Molecular mechanisms were investigated through multiple experimental approaches: the interaction between NRF2 and the DUSP1 promoter was examined using dual fluorescence reporter assays, while the activation status of CEBPB, NRF2, DUSP1, antioxidant proteins, and MAPK pathway components was analyzed via immunofluorescence and western blotting. Our findings demonstrate that CEBPB overexpression significantly enhanced cellular proliferation, clone formation, cell cycle progression, and antioxidant capacity, while simultaneously reducing apoptosis rates and reactive oxygen species (ROS) levels. Conversely, CEBPB knockdown or NRF2 inhibition produced opposing effects. These results establish that CEBPB-mediated NRF2 activation promotes OC cell proliferation and antioxidant defense mechanisms. Mechanistically, we identified that NRF2 directly binds to the DUSP1 promoter, as confirmed by dual-luciferase reporter assays. NRF2 activation led to upregulation of DUSP1 and phosphorylated ERK1/2 levels, while downregulating JNK and p38 phosphorylation. These findings were further validated in vivo, confirming that CEBPB activates NRF2 to regulate the MAPK pathway through DUSP1, thereby promoting OC cell proliferation and antioxidant capacity. In conclusion, our study reveals a novel regulatory axis in which CEBPB activates NRF2 to regulate the MAPK pathway via DUSP1, driving malignant progression and enhancing antioxidant activity in OC.
Growth Hormone-Releasing Hormone and Somatostatin exert opposing activities in Growth Hormone (GH) regulation. Herein - and based on recent findings - we provide our insights on the potential therapeutic role of GH suppr...Growth Hormone-Releasing Hormone and Somatostatin exert opposing activities in Growth Hormone (GH) regulation. Herein - and based on recent findings - we provide our insights on the potential therapeutic role of GH suppression on endothelium-dependent disorders.
FAM3 metabolism-regulating signaling molecule A (FAM3A) is a mitochondrial protein belonging to the gene family with the potential for the treatment of ischemic diseases. FAM3A promotes adenosine triphosphate (ATP) prod...FAM3 metabolism-regulating signaling molecule A (FAM3A) is a mitochondrial protein belonging to the gene family with the potential for the treatment of ischemic diseases. FAM3A promotes adenosine triphosphate (ATP) production and improves mitochondrial function by increasing ATP synthase activity and activating the protein kinase B-cyclic AMP-responsive element binding protein-forkhead box D3-ATP synthase regulatory loop, thereby reducing reactive oxygen species production and inhibiting oxidative stress-induced cell death. Additionally, FAM3A activates the nuclear factor erythroid 2-related factor 2 signaling pathway and upregulates the expression of antioxidant proteins, further enhancing the cellular oxidative defense capacity. During angiogenesis, FAM3A positively regulates vascular endothelial growth factor A and promotes endothelial cell migration, proliferation, and tube formation. FAM3A is closely related to atherosclerosis, ischemic encephalopathy, liver ischemia - reperfusion injury, myocardial ischemia, and acute kidney injury. FAM3A plays a role in the course of these diseases via multiple mechanisms, including the phosphatidylinositol 3-kinase/protein kinase B signaling pathway, effectively reducing the inflammatory response and oxidative stress, and influencing disease development. This review comprehensively examines the role of FAM3A in the pathophysiological processes of ischemic diseases across various organs.
SKP2, an E3 ubiquitin ligase component of the SCF complex, plays a critical role in cell cycle regulation by targeting key inhibitors like p27, p21, and p57 for degradation, thereby promoting G1-S transition. Its overexp...SKP2, an E3 ubiquitin ligase component of the SCF complex, plays a critical role in cell cycle regulation by targeting key inhibitors like p27, p21, and p57 for degradation, thereby promoting G1-S transition. Its overexpression is strongly associated with urological malignancies, including prostate, bladder, and kidney cancers, where it correlates with aggressive disease and poor prognosis. SKP2 drives tumor progression, via enhancing cancer cell proliferation, invasion, and metastasis. Targeting SKP2 through small molecule inhibitors or combination therapies holds promise for cancer treatment. However, challenges remain, including understanding its role in cancer stem cells, metastasis, and treatment resistance. Continued research is essential to harness SKP2's potential as a therapeutic target and biomarker for personalized medicine in urological cancers.
Ultraviolet radiation is a major factor in causing skin aging. Compared to younger individuals, older adults exhibit a significant imbalance in the M1/M2 macrophage ratio, with an elevated proportion of M1 macrophages, b...Ultraviolet radiation is a major factor in causing skin aging. Compared to younger individuals, older adults exhibit a significant imbalance in the M1/M2 macrophage ratio, with an elevated proportion of M1 macrophages, but little is known about the role of macrophages in skin aging. Here, we report the critical role of M2 macrophages and PKM2 in preventing fibroblast photoaging. UVB-treated photoaged fibroblasts showed a reduction in PKM2. Compared to M1 macrophages, treatment with M2 macrophage significantly alleviated this photoaging and enhanced PKM2 synthesis in fibroblasts. Mechanistically, this is due to the secretion of CCL1 by M2 macrophages, which acts on the CCR8 receptor on the cell surface, promoting PKM2 production in photoaged fibroblasts. This further activates the TGF-β1/Smad2 pathway, thereby reducing cellular aging. This provides a potential strategy for the treatment of skin photoaging.
Patients frequently experience physical, mental, and even financial distress because of acute or chronic skin wounds. In severe situations, scarring on the skin can be quite noticeable, cause persistent discomfort, restr...Patients frequently experience physical, mental, and even financial distress because of acute or chronic skin wounds. In severe situations, scarring on the skin can be quite noticeable, cause persistent discomfort, restrict joint motion, or be mentally taxing. Hair-follicle-associated pluripotent (HAP) stem cells were discovered by our laboratory, in the bulge area of the hair follicle and can differentiate to neurons, glia, beating cardiomyocytes, keratinocytes and nascent vessels. In the present study, HAP stem cell sheets were formed by culturing the upper part of hair follicles and implanting into mice with skin ulcers. The HAP stem cell sheets contained keratinocytes, endothelial cells and neurons. Autologous HAP stem cell sheet implantation to the dorsal wound in C57BL/6J mice significantly accelerated wound closure compared with non-implanted control mice. HAP-stem-cell sheets expressing green fluorescent protein (GFP) implanted into nude mice differentiated into keratinocytes in the epidermis, and neurons and endothelial cells in the dermis. The thicknesses of the epidermis and dermis and M2 macrophage and myofibroblast infiltration into the wound were significantly decreased in HAP-stem cell-implanted mice compared with non-implanted control mice. Expression levels of TGF-β1, COL1A2 and COL3A1 mRNA in the wound were significantly decreased in HAP stem cell-implanted mice compared with non-implanted control mice. These results suggest that implanting HAP stem cell sheets accelerates cutaneous wound closure and suppresses scar formation. The HAP stem cells used in the present study thus have potential as a future clinical strategy for accelerating wound healing.
The cell-cycle regulated methyltransferase SET8 is the sole enzyme responsible for the mono-methylation of histone H4 at lysine 20 (H4K20) that is the substrate for di- and trimethylation mainly by SUV4-20Hs enzymes. Bot...The cell-cycle regulated methyltransferase SET8 is the sole enzyme responsible for the mono-methylation of histone H4 at lysine 20 (H4K20) that is the substrate for di- and trimethylation mainly by SUV4-20Hs enzymes. Both SET8 and SUV4-20Hs have been implicated in regulating DNA repair pathway choice through the inverse affinities of BRCA1-BARD1 and 53BP1 complexes for disparate methylation states of H4K20. However, the precise and respective functions of each H4K20 methyltransferase in DNA repair pathways remain to be clarified. Here, we show that SET8 acts as a potent chromatin inhibitor of homologous recombination and that its timely degradation during DNA replication is essential for the spontaneous nuclear focal accumulation of BRCA1 and RAD51 complexes during the S phase. Strikingly, the anti-recombinogenic function of SET8 is independent of SUV4-20H activity but requires the subsequent recruitment of the ubiquitin ligase RNF168. Moreover, we show that SET8-induced BRCA1 inhibition is not necessarily related to the loss of BARD1 binding to unmethylated histone H4K20. Instead, it is largely caused by the accumulation of 53BP1 in a manner depending on the concerted activities of SET8 and RNF168 on chromatin. Conversely, the lack of SET8 and H4K20 mono-methylation on newly assembly chromatin after DNA replication led to the untimely accumulation of BRCA1 on chromatin at the subsequent G1 phase. Altogether, these results establish the activity of SET8 on chromatin as a primordial epigenetic lock of the BRCA1-mediated HR pathway during the cell cycle.
Elevated succinate accumulation has been demonstrated to be associated with metabolic and inflammatory disorders. Our previous study revealed that adipose-derived stem cells (ADSC) from obese individuals exhibit high suc...Elevated succinate accumulation has been demonstrated to be associated with metabolic and inflammatory disorders. Our previous study revealed that adipose-derived stem cells (ADSC) from obese individuals exhibit high succinate, reduced biological activity, and mitochondrial dysfunction. However, the precise role of succinate in these processes remains unclear. Here, we investigated the effects of excess succinate on cellular biological activity, immunomodulatory capacity, and mitochondrial function of ADSC. We found that elevated succinate levels in ADSC decreased proliferation and differentiation potential, while promoting M1 macrophage polarization. Furthermore, succinate accumulation impaired mitochondrial biogenesis and metabolism, increasing in reactive oxygen species (ROS) production and inflammatory responses. Transcriptome sequencing analysis further confirmed that succinate upregulated inflammatory pathways, suppressed mitochondrial biogenesis and metabolism, and enhanced cellular apoptosis and senescence, accompanied by reduced DNA replication and repair. Overall, these findings imply that succinate accumulation in ADSC triggers inflammatory response and mitochondrial dysfunction, potentially contributing to a decline of cellular biological activity. Targeting succinate may offer therapeutic potential for metabolic disorders.
Microtubules are polymers of α/β tubulin dimers that build the mitotic spindle, which segregates duplicated chromosomes during cell division. Microtubule function is governed by dynamic instability, whereby cycles of gro...Microtubules are polymers of α/β tubulin dimers that build the mitotic spindle, which segregates duplicated chromosomes during cell division. Microtubule function is governed by dynamic instability, whereby cycles of growth and shrinkage contribute to the forces necessary for chromosome movement. Regulation of microtubule growth velocity requires cell cycle-dependent changes in expression, localization and activity of microtubule-associated proteins (MAPs) as well as tubulin post-translational modifications that modulate microtubule dynamics. It has become clear that optimal microtubule growth velocities are required for proper chromosome segregation and ploidy maintenance. Suboptimal microtubule growth rates can result from altered activity of MAPs and could lead to aneuploidy, possibly by disrupting the establishment of microtubule bundles at kinetochores and altering the mechanical forces required for sister chromatid segregation. Future work using high-resolution, low-phototoxicity microscopy and novel fluorescent markers will be invaluable in obtaining deeper mechanistic insights into how microtubule processes contribute to chromosome segregation.