Knee osteoarthritis (KOA) is a degenerative joint disorder characterized by articular cartilage degeneration and synovial inflammation. Dysfunction mediated by autophagy-related protein 5 (ATG5) represents a key driver o...Knee osteoarthritis (KOA) is a degenerative joint disorder characterized by articular cartilage degeneration and synovial inflammation. Dysfunction mediated by autophagy-related protein 5 (ATG5) represents a key driver of KOA pathogenesis, while the cGAS-STING-NLRP3 signaling axis is closely associated with inflammation and apoptosis. Therefore, this study aims to explore the regulatory effect of ATG5 on the cGAS-STING-NLRP3 axis and its specific role in KOA. HE staining, Safranin O-fast green staining were used to assess the degree of cartilage degeneration in KOA rats. TUNEL staining were applied to observe the apoptosis of chondrocytes in cartilage tissues. Levels of inflammatory factors in serum of rats and cells were detected by ELISA. Expression of proteins levels was analyzed using western blot and immunofluorescence assay. Flow cytometry was used to investigate the apoptosis. In cartilage tissues of KOA rats, ATG5 was lowly expressed, while cGAS, STING and NLRP3 were overexpressed. Co-localization was observed between ATG5 and STING. Overexpression of ATG5 led to decreased expression of inflammatory factors, cGAS, STING, NLRP3, p62 and Cleaved caspase-1, a reduced Mankin score, and increased Beclin1 expression. However, knockout of ATG5 reversed these changes. Additionally, overexpression of ATG5 decreased the apoptosis rate of chondrocytes, whereas inhibition of ATG5 promoted chondrocyte apoptosis. In conclusion, ATG5 regulates the cGAS-STING-NLRP3 axis, thereby promoting chondrocyte autophagy and inhibiting inflammation, which in turn protects chondrocytes and alleviates KOA. In conclusion, ATG5 modulates the cGAS-STING-NLRP3 axis, thereby enhancing chondrocyte autophagy and suppressing inflammation, which collectively protects chondrocytes and alleviates the progression of KOA.
Bone fracture healing is a complex physiologic process that aims at restoring the damaged bone to its pre-injury state and cellular composition. Exosomes secreted by bone marrow mesenchymal stem cells (BMSCs) are emergin...Bone fracture healing is a complex physiologic process that aims at restoring the damaged bone to its pre-injury state and cellular composition. Exosomes secreted by bone marrow mesenchymal stem cells (BMSCs) are emerging as a promising strategy to promote bone regeneration due to exosomal bioactive cargos. Furthermore, N6-Methyladenosine (m6A) methylation affects osteoblastic differentiation and bone remodeling. This study is designed to clarify the role and mechanism of BMSC-derived exosomal Methyltransferase-like 14 (METTL14) in osteogenesis. METTL14 and bone morphogenetic protein 2 (BMP2) levels were detected by RT-qPCR. METTL14, exosome-specific markers, BMP2, and IGF2BP1 protein levels were determined using Western blot. Cell viability, proliferation, and apoptosis were examined using MTT, EdU, and flow cytometry. The degree of osteogenic differentiation was verified by the Alizarin Red S staining assay and ALP activity assay. The interaction between METTL14 and BMP2 was analyzed using methylated RNA immunoprecipitation (MeRIP)-qPCR and RIP assays. METTL14 and BMP2 levels were decreased in delayed fracture healing (DFH), a common complication after fracture surgery. METTL14 upregulation expedited MC3T3-E1 cell viability, proliferation, and repressed apoptosis. METTL14 promotes osteogenic differentiation of MC3T3-E1 cells by enhancing ALP activity and mineralized formation. After co-culturing BMSC-derived exosomes and MC3T3-E1 cells, BMSC-derived exosomal METTL14 expedited the osteoblast activity. Mechanistically, METTL14 stabilized BMP2 mRNA through the m6A-IGF2BP1-dependent mechanism. These findings indicated that BMSC-derived exosomes encapsulate METTL14 and transport it into MC3T3-E1 cells, and the transported METTL14 could accelerate the osteogenesis by regulating the stability of BMP2 mRNA, which provided a potentially effective therapeutic strategy for bone regeneration.
Retinopathy of prematurity (ROP) is an abnormal proliferative disease of retinal blood vessels. This study investigates the role of Robo4 in angiogenesis and its molecular mechanisms in the Oxygen-Induced Retinopathy (OI...Retinopathy of prematurity (ROP) is an abnormal proliferative disease of retinal blood vessels. This study investigates the role of Robo4 in angiogenesis and its molecular mechanisms in the Oxygen-Induced Retinopathy (OIR) model and a hypoxic injury model of human umbilical vein endothelial cells (HUVECs). In the OIR model, Roundabout 4 (Robo4) expression was significantly decreased, while Robo4 overexpression inhibited neovascularization and alleviated retinal tissue damage. Under hypoxic injury-inducing conditions in HUVECs, Robo4 expression was inhibited, but overexpression enhanced cell proliferation and inhibited angiogenesis by negatively regulating the expression of adenosine diphosphate ribosylation factor 6 (ARF6) and vascular endothelial growth factor (VEGF). Molecular analysis revealed that Robo4 regulates VEGF expression through ARF6. Overexpression of Robo4 reduced ARF6 and VEGF expression, while knocking down Robo4 increased ARF6 and VEGF levels, confirming Robo4's role as a key regulator in the ARF6-VEGF axis. Additionally, Robo4 overexpression significantly inhibited the angiogenic capacity of HUVECs, and ARF6 overexpression partially reversed Robo4-mediated inhibition of angiogenesis. In the OIR model, Robo4 overexpression significantly suppressed pathological neovascularization, as evidenced by a significant reduction in the percentage of non-perfused areas. Furthermore, Robo4 overexpression downregulated ARF6 and VEGF expression levels and promoted the normalization and remodeling of retinal vascular structures. These findings suggest that Robo4 inhibits pathological angiogenesis by regulating ARF6 and VEGF, providing insights into its potential as a therapeutic target for retinal diseases like ROP.
Diffuse large B-cell lymphoma (DLBCL) is a malignant tumor and research on its therapeutic targets has received increasing attention. It has been reported that Leucyl-tRNA synthetase (LARS) contributes to the growth and...Diffuse large B-cell lymphoma (DLBCL) is a malignant tumor and research on its therapeutic targets has received increasing attention. It has been reported that Leucyl-tRNA synthetase (LARS) contributes to the growth and migration of non-Hodgkin lymphoma (NHL), yet its effect on DLBCL progression remains to be elucidated. MTT and flow cytometry were carried out to determine the cellular phenotypes of DLBCL cells under LARS overexpression. The differentially expressed proteins (DEPs) were screened by mass spectrometry. ELISA, western blot, and xenograft tumor experiments were implemented to confirm the impact of LARS and the LRPPRC/HIF-1α axis on malignant progression and glycolysis. Dual-luciferase reporter assay and chromatin immunoprecipitation (ChIP) were applied for exploring the underlying mechanism by which LARS was regulated. LARS promoted the malignant phenotypes, such as increasing cell proliferation and inhibiting apoptosis, and enhanced abnormal glycolysis both in vitro and in vivo. Based on mass spectrometry and functional recovery experiments, we found that LARS upregulated LRPPRC expression. More importantly, overexpressing LRPPRC facilitated malignant phenotypes and glycolysis through the elevation of HIF-1α expression, which could be reversed by silenced HIF-1α. Mechanistically, LARS promoted the expression of HIF-1α by activating LRPPRC, which in turn enhanced the transcription of HK2, thereby facilitating malignant progression and abnormal glycolysis. LARS facilitated abnormal glycolysis via the LRPPRC/HIF-1α/HK2 axis, thereby promoting the malignant progression of DLBCL.
Colorectal cancer, a prevalent digestive system malignancy, is characterized by a high incidence, low early detection rate, limited surgical resection opportunities, and high mortality. Palmatine (PAL), an active ingredi...Colorectal cancer, a prevalent digestive system malignancy, is characterized by a high incidence, low early detection rate, limited surgical resection opportunities, and high mortality. Palmatine (PAL), an active ingredient primarily found in Coptidis Rhizoma, exhibits diverse pharmacological effects, including antibacterial, anti-inflammatory, and anti-tumor properties. While PAL has been shown to effectively curb the progression of colorectal cancer, the underlying mechanisms have yet to be fully elucidated. In this study, we demonstrated the inhibitory effect of PAL on colorectal cancer growth via the miR-363-3p/AURKA axis, utilizing an AOM/DSS-induced colorectal cancer model in C57BL/J mice, a subcutaneous tumor xenograft model in nude mice, and in vitro assays using HCT-116 and SW620 cells. PAL upregulates miR-363-3p expression, promotes the interaction between AURKA 3'UTR mRNA and miR-363-3p, impedes AURKA mRNA translation into AURKA protein, thereby inhibiting colorectal cancer cell proliferation and migration, and suppressing the initiation and progression of colorectal cancer. These results expand the understanding of the regulatory mechanisms by which PAL influences colorectal cancer development, and may provide new potential targets for colorectal cancer diagnosis and therapy.
Mersakova S, Marcinek J, Brany D
… +31 more, Chodelkova O, Vorcak M, Cermak M, Poturnajova M, Kozovska Z, Skovierova H, Novakova S, Mitruskova B, Loderer D, Grendar M, Holubekova V, Hornakova A, Melegova J, Brozmanova M, Palkoci B, Vojtko M, Kycina R, Pindura M, Janik J, Mikolajcik P, Gabonova E, Laca L, Nicodemou A, Danisovic L, Plank L, Halasova E, Mraz M, Matuskova M, Lasabova Z, Kalman M, Strnadel J
Metastatic colorectal cancer is one of the most critical causes of cancer-related dead in patients suffering this type of malignancy. As the liver metastases are found in almost 25-50% of all colorectal cancer patients,...Metastatic colorectal cancer is one of the most critical causes of cancer-related dead in patients suffering this type of malignancy. As the liver metastases are found in almost 25-50% of all colorectal cancer patients, there is an urgent need for suitable in vitro and in vivo models. To the best of our knowledge, only a limited number of well-described 2D or 3D organoid/spheroid cell lines originated from human liver metastases was reported for this type of malignancy and submitted to ATCC and other repository centers for general use. Here, we present a novel, well-characterized cell line Coala that was derived directly from liver metastasis of patient with colorectal cancer. Coala cancer cell line was derived by 2D and 3D culture technology and can be, therefore, cultured as standard 2D cell line or as 3D spheroid culture. When transplanted into athymic mice, Coala cell line forms fast growing tumors with histological features similar to original tumor and expressed CDX2, SATB2, and CK20. Whole exome sequencing analysis of Coala cell line showed the presence of pathogenic mutations in C8B, PRKCQ, IRGM, FGFR4, POLR1C, APC and TP53 genes. As verified by antibody array chip analysis, Coala cells express Snail, Pdx-1, FoxA2, and E-cadherin. Our cell line represents a new, well characterized in vitro research tool for cancer research and will be available to researchers in the field via biorepository centers.
The abundance of polyploid giant cancer cells (PGCCs) is correlated with malignant tumor progression. PGCCs divide asymmetrically to generate daughter cells with high invasive and migratory capacities. The objective of t...The abundance of polyploid giant cancer cells (PGCCs) is correlated with malignant tumor progression. PGCCs divide asymmetrically to generate daughter cells with high invasive and migratory capacities. The objective of this study was to investigate the mechanism by which the overexpression of polo-like kinase 1 (PLK1) regulates the migration and invasion of PGCCs and their daughter cells (PDCs). In the present study, PGCC formation was induced by CoCl in Hct116 and LoVo cells. The mRNA expression and protein levels were detected using reverse transcription‒quantitative PCR and western blotting, respectively. The regulatory relationships among related proteins were determined via small molecule inhibitors, transient small interfering RNA (siRNA) transfection, and coimmunoprecipitation. Our findings indicated that CoCl induces PGCC formation and produces progeny cells with increased invasive and metastatic abilities. PLK1 was expressed in PDCs and showed altered subcellular localization. In addition, aurora kinase A (AURKA) activated PLK1 by phosphorylating it at serine 137 and threonine 210. Activated PLK1 first stabilizes β-catenin accumulation in the cytoplasm and subsequently promotes its entry into the nucleus. After entering the nucleus, β-catenin promotes the accumulation of the tyrosine 705 site of signal transducer and activator of transcription 3 (pSTAT3-Tyr705), which promotes the downstream expression of target proteins, such as cyclin D1, matrix metallopeptidase 2 (MMP2) and c-MYC, and EMT-associated proteins as transcription factors. Furthermore, STAT3 and PLK1 form a feedback loop, allowing STAT3 to regulate PLK1 expression. In conclusion, PLK1 phosphorylation at different sites regulates the transcriptional function of pSTAT3-Tyr705 by promoting β-catenin nuclear translocation. Furthermore, AURKA and STAT3 affected PLK1 expression.
Anaplastic large-cell lymphoma (ALCL) accounts for 15% of all peripheral T-cell lymphomas globally and can be further divided into subcategories, of which patients with ALK-negative ALCL have dismal prognosis and overall...Anaplastic large-cell lymphoma (ALCL) accounts for 15% of all peripheral T-cell lymphomas globally and can be further divided into subcategories, of which patients with ALK-negative ALCL have dismal prognosis and overall survival. We established a patient-derived xenograft (PDX) and in vitro model (designated PTCL-S1) of TP63-rearranged ALK-negative ALCL from the primary tumour site of a 55-year old Chinese woman. Whole genome sequencing of the patient's tumour identified various mutations including AKT1 and NOTCH1, as well as the TP63-TBL1XR1 gene fusion. RNA sequencing followed by Sanger sequencing confirmed the gene rearrangement in original tumour, PDX and PTCL-S1 cell line. Immunohistochemistry profiling of the PDX model and cell-line were consistent with the patient's primary tumour sample (CD3 + /CD30 + /CD79a-). Cytotoxic agents (doxorubicin, etoposide and gemcitabine) commonly used in ALCL treatment exhibited potent anti-proliferative activity in the cell-line. In conclusion, the established PTCL-S1 cell line can be a useful tool for further investigation of the understanding of TP63-rearranged ALK-negative ALCL.
Pleomorphic rhabdomyosarcoma (pRMS) is a rare and highly malignant mesenchymal tumor. Complete resection is the only curative treatment available, owing to the limited efficacy of chemotherapy and radiotherapy. Therefore...Pleomorphic rhabdomyosarcoma (pRMS) is a rare and highly malignant mesenchymal tumor. Complete resection is the only curative treatment available, owing to the limited efficacy of chemotherapy and radiotherapy. Therefore, developing novel therapies for pRMS is important for improving clinical outcomes. Herein, a novel pRMS cell line, SMU-pRMS, was established for a detailed understanding of the biological characteristics of pRMS, thereby developing new therapies. A tissue sample from a surgically resected tumor of a 69-year-old patient was subjected to primary culture. The cell line was established and authenticated by evaluating the short tandem repeats of DNA microsatellites. Monolayer cultures of SMU-pRMS cells exhibited constant growth, spheroid formation, and invasiveness. These cells exhibited high chemosensitivity to eribulin. In addition, mice inoculated with SMU-pRMS cells developed tumors after 4 weeks. Therefore, the SMU-pRMS cell line is a useful tool for investigating pRMS development and evaluating novel therapeutic agents.
Functional exhaustion of T lymphocytes is considered an important factor in the failure of tumor treatment. Accumulating evidence has shown that tumor cells cultivate their immune microenvironment through secreting exoso...Functional exhaustion of T lymphocytes is considered an important factor in the failure of tumor treatment. Accumulating evidence has shown that tumor cells cultivate their immune microenvironment through secreting exosomes. However, the mechanism through which tumor cell-derived exosomes participate in the regulation of lymphocyte function remains unclear. In this study, we found that exosomes derived from prostate cancer (PCa) cells were able to upregulate the expression of PD-1 and TIM-3 in CD8 T cells, inducing the secretion of cytokines related to T cell exhaustion and significantly decreasing the ability to kill PCa cells. Importantly, our data indicated that treatment with GW4869 could reverse the effects of PCa-derived exosomes on CD8 T cells and further inhibit the growth of PCa cells in vivo and in vitro by blocking the generation of exosomes. Our findings support the notion that exosomes derived from PCa cells can induce T cell exhaustion and promote PCa progression, while treatment of GW4869 effectively rejuvenates CD8 T cells and reverses the effect of PCa exosomes. These findings indicate that GW4869 has potential in the treatment of PCa.
Periodontal ligament stem cells (PDLSCs) constitute a promising source for successful periodontal regeneration. This study aims to explore roles of metformin, krüppel-like factor 2 (KLF2), and miR-181a-5p in mediating os...Periodontal ligament stem cells (PDLSCs) constitute a promising source for successful periodontal regeneration. This study aims to explore roles of metformin, krüppel-like factor 2 (KLF2), and miR-181a-5p in mediating osteogenic differentiation of human PDLSCs (hPDLSCs) following lipopolysaccharide (LPS) stimulation. The osteogenic differentiation potential of hPDLSCs isolated from human premolar root samples were examined by alkaline phosphatase (ALP) staining, ALP activity assay, Alizarin red S staining, and Western blotting of osteogenic markers. Metformin pretreatment at dose of 100 μM significantly resulted in increased ALP activity, elevated protein expressions of osteogenic markers, and more generated mineralized matrix in hPDLSCs with LPS stimulation. KLF2 and miR-181a-5p were found to be increased by metformin pretreatment at dose of 100 μM in hPDLSCs with stimulation but not in hPDLSCs without LPS stimulation. The interaction between the KLF2 and the promoter of miR-181a-5p was noted by the dual-luciferase reporter assay. KLF2 knockdown or miR-181a-5p inhibition notably abrogated the improvements of osteogenic differentiation by metformin pretreatment in LPS-stimulated hPDLSCs. The findings of the study indicate metformin protects hPDLSCs against impaired osteogenic differentiation of hPDLSCs after LPS stimulation by KLF2-mediated activation of miR-181a-5p under inflammation conditions.
Severe sepsis can escalate to acute lung injury (ALI), a critical condition with limited treatment options. Emerging research highlights the role of non-coding RNAs, including microRNAs (miRNAs) and circular RNAs (circRN...Severe sepsis can escalate to acute lung injury (ALI), a critical condition with limited treatment options. Emerging research highlights the role of non-coding RNAs, including microRNAs (miRNAs) and circular RNAs (circRNAs), in regulating epithelial and immune cell responses in sepsis-induced ALI. This study delves into the circAGFG1/miR-195-5p/PD-L1 axis to elucidate its potential in this context. The conducted experiments using Th17 cells differentiated from CD4+ T cells isolated from human umbilical cord blood, co-cultured with transfected Calu-3 cells. The assessed cell viability, RNA, and protein expression levels under inflammatory stimuli. In addition, we collected bronchoalveolar lavage fluid and lung tissues from a cecal ligation and puncture mouse model. Quantification of RNA and protein expression of inflammatory and survival markers provided insights into the regulatory mechanisms involved. Peripheral blood samples from sepsis/ALI patients and healthy controls revealed that circAGFG1 expression is significantly downregulated, while miR-195-5p expression is upregulated in patients compared to healthy individuals. In both Calu-3 cells and induced Th17 cells, as well as in sepsis-induced ALI mouse models, the circAGFG1/miR-195-5p/PD-L1 axis was shown to regulate the expression of inflammatory cytokines, Th17 cell differentiation, and markers of epithelial cell survival. Results show the circAGFG1/miR-195-5p/PD-L1 axis plays a critical role in the pathogenesis of sepsis-induced ALI. Targeting this axis could present a novel therapeutic strategy for preventing ALI in sepsis and other conditions characterized by heightened lung inflammation. This research paves the way for new treatment approaches, offering hope for improved outcomes in patients suffering from severe sepsis and ALI.
Giant cell tumor of bone (GCTB) is locally aggressive and rarely metastasizing mesenchymal tumor, molecularly characterized by the presence of H3-3A mutation. The management of GCTB is problematic because of local recurr...Giant cell tumor of bone (GCTB) is locally aggressive and rarely metastasizing mesenchymal tumor, molecularly characterized by the presence of H3-3A mutation. The management of GCTB is problematic because of local recurrence after curative surgical treatment, and complex biological and molecular backgrounds of etiology and disease progression. Development of multidisciplinary therapy has been required in GCTB, and targeting treatments against nuclear factor kappa-B ligand and epidermal growth factor receptor to neoplastic stromal cells were applied to the clinical practice. To promote the translational research of GCTB, we developed two cell lines from primary tumor tissues. The established cell lines exhibited H3-3A gene mutations: NCC-GCTB11-C1 (p.Gly35Leu), and NCC-GCTB12-C1 (p.Gly35Trp). The p.Gly35Leu (G35L) is a rare variant, and NCC-GCTB11-C1 is the first GCTB cell line holding it. The two cell lines showed constant proliferation, spheroid formation, and invasion, making them suitable for in vitro studies. We demonstrated that the two cell lines were useful for drug screening using 221 anti-cancer agents. NCC-GCTB11-C1 and NCC-GCTB12-C1 will be critical resources for the development of novel targeted treatments by their application for drug screening.
Dental pulp stem cells (DPSCs) are a population of adult stem cells with self-renewal capacity and multilineage differentiation potential, widely utilized in tissue engineering and regenerative medicine. However, with in...Dental pulp stem cells (DPSCs) are a population of adult stem cells with self-renewal capacity and multilineage differentiation potential, widely utilized in tissue engineering and regenerative medicine. However, with increasing donor age or prolonged in vitro expansion, DPSCs gradually exhibit senescence-associated phenotypes, including reduced proliferative capacity and impaired differentiation potential. This review comprehensively summarizes current advances in the study of DPSCs senescence. First, it explores how the aging dental pulp microenvironment influences the biological characteristics of DPSCs. It then focuses on the interplay between DPSCs senescence and mitochondrial dysfunction, epigenetic alterations, as well as key signaling pathways such as p53/p21/p16. On this basis, recent molecular strategies for delaying DPSCs senescence are discussed, such as melatonin can alleviate DPSCs senescence by inhibiting the expression of matrix metalloproteinase, and metformin could interfere DPSCs senescence by AMPK/mTOR signaling pathway, etc. Lastly, a comparative analysis of senescence characteristics among DPSCs, bone marrow and adipose derived mesenchymal stem cells, is conducted to contextualize the unique and shared features of these cell types. Collectively, this review could provide a theoretical foundation and practical insights for advancing anti-senescence strategies in DPSC-based regenerative applications.
Monkeypox is a viral zoonotic infection that has emerged as a significant public health threat recently. The world has experienced a global outbreak of the Monkeypox virus (MPXV), with 124,753 confirmed cases in 128 coun...Monkeypox is a viral zoonotic infection that has emerged as a significant public health threat recently. The world has experienced a global outbreak of the Monkeypox virus (MPXV), with 124,753 confirmed cases in 128 countries and a total of 272 fatalities as of February 13, 2025. This alarming increase in cases demands immediate attention to the underlying causes of the surge. The World Health Organization (WHO) has endorsed the use of the MVA-BN vaccine, which was previously approved for smallpox prevention. However, there are currently no antiviral treatments approved by the FDA for MPXV. In addition, the emergence of mutations in MPXV strains poses challenges for the development and effectiveness of viable vaccines. Developing conventional vaccines is typically expensive, time-consuming, and requires extensive validation processes. Moreover, these vaccines often demonstrate suboptimal efficacy against emerging viral strains. As a response to these challenges, multi-epitope vaccines have gained significant interest for their potential to activate B-cell and T-cell responses, providing prolonged immunological activity against pathogens. These vaccines feature a targeted approach, offering chemical stability, non-infectious properties, rapid and cost-effective production, enhanced safety, and the potential to elicit a robust immune response. Several studies employing immunoinformatics have confirmed the efficacy of multi-epitope vaccines, designed to provide comprehensive immune protection against MPXV. These vaccines promise to deliver a strong immune response, better coverage against various viral strains and variants, and improved stability and effectiveness. In addition, they are expected to be non-allergenic, non-toxic, and highly antigenic. While promising results have been reported regarding the use of these vaccines for monkeypox, further research on their efficacy, delivery systems, and additional preclinical and clinical trials is crucial to maximize their benefits for humanity.
Primary ovarian squamous cell carcinoma (POSCC) represents an exceedingly rare subtype of epithelial ovarian cancer (EOC) characterized by a cryptic etiology and insidious onset. The rarity and high mortality associated...Primary ovarian squamous cell carcinoma (POSCC) represents an exceedingly rare subtype of epithelial ovarian cancer (EOC) characterized by a cryptic etiology and insidious onset. The rarity and high mortality associated with pure primary ovarian squamous cell carcinoma (SCC) make it challenging to conduct large randomized controlled studies. Furthermore, there are currently no commercially available ovarian SCC cell lines for research purposes, necessitating the urgent establishment of novel lines. To our knowledge, this study reports the first preclinical model of primary ovarian squamous cell carcinoma (POSCC), denoted as ZOC254. We have detailed the establishment and characterization of ZOC254, derived from a 64-year-old female patient, which preserves the primary tumor's original traits across various levels during prolonged in vitro expansion. Whole-exome sequencing (WES) of both the primary tumor and derived cell line revealed homologous recombination deficiency (HRD) and high tumor mutational burden (TMB). ZOC254 also exhibited the PIK3CA: p.E542K mutation associated with targeted therapy. The effectiveness of olaparib, everolimus, and conventional chemotherapeutic agents for ovarian cancer was preliminarily assessed on the growth of patient-derived cells (PDC). The POSCC cell line and derived xenograft transplantation model reported in this study serve the purpose of broadening the resources accessible for preclinical investigations into ovarian squamous carcinoma.
Human umbilical cord mesenchymal stromal cells (hucMSCs) have emerged as a promising therapeutic option for acute liver failure (ALF). However, the detailed mechanism by which hucMSCs modulate hepatocyte pyroptosis in AL...Human umbilical cord mesenchymal stromal cells (hucMSCs) have emerged as a promising therapeutic option for acute liver failure (ALF). However, the detailed mechanism by which hucMSCs modulate hepatocyte pyroptosis in ALF remains unclear. In this study, we induced ALF in mice using acetaminophen (APAP). Mice were intravenously injected with 1 × 10 hucMSCs/ hucMSCs-Exo via the tail vein 6 h after APAP administration. Liver pathological changes were assessed by hematoxylin and eosin (H&E) staining. Subsequently, an in vitro model of liver cell failure and pyroptosis model was established using LO2 cells treated with lipopolysaccharide (LPS) and adenosine 5'-triphosphate (ATP). The levels of cell pyroptosis marker caspase-1 were detected by flow cytometry analysis, RT-qPCR assay, and western blot assay. The study employed a comprehensive approach, including flow cytometry analysis, RT-qPCR assay, miRNA sequencing, and luciferase reporter gene experiments. hucMSCs and hucMSCs- exosomes (MSCs-Exo) inhibited cell inflammation to improve ALF in vivo model of ALF and inhibited hepatocyte pyroptosis in LO2 cells induced by ATP and LPS. MiR-423-5p emerged as a potential mediator of the anti-pyroptotic effects of hucMSCs-Exo, with ZBP1 identified as one of its downstream targets. Subsequent validation confirmed that miR-423-5p targets ZBP1 to regulate pyroptosis. These findings highlight the role of hucMSCs-derived exosomal miR-423-5p in inhibiting hepatocyte pyroptosis in LO2 cells induced by ATP and LPS. miR-423-5p serves as a crucial mediator in this process, targeting ZBP1, a protein significantly involved in the pyroptotic pathway. Our findings may provide basics for further research on mechanism of hucMSCs and present a promising cell-free strategy treating ALF.
Dysregulation of circRNAs has been found to engage in the progression of many hematologic malignancies including acute myeloid leukemia (AML). In this study, we identified significantly downregulated circTADA2A, derived...Dysregulation of circRNAs has been found to engage in the progression of many hematologic malignancies including acute myeloid leukemia (AML). In this study, we identified significantly downregulated circTADA2A, derived from linear transcriptional adaptor 2A (TADA2A) gene, in AML associated circRNAs microarrays using GEO2R tool. We aimed to elucidate the roles of circTADA2A in AML and the mechanisms involved. Quantitative reverse transcription PCR was used for verification of circTADA2A levels in AML specimens, and its diagnostic value and clinical significance were assessed. The effects of circTADA2A on proliferation and ferroptosis on THP-1 and HL-60 cells were carried out using cell counting kit-8, 5'-ethynyl-2'-deoxyuridine, Fe, lipid reactive oxygen species (ROS) and malondialdehyde (MDA) assays. Luciferase reporter, fluorescence in situ hybridization, RNA immunoprecipitation, and RNA pull-down assays were implemented to investigate the potential miRNAs that mediated circTADA2A functioning. We confirmed that circTADA2A levels were lowly expressed in plasma and bone marrow of AML patients, and associated with bone marrow blasts and cytogenetic risk. Plasma circTADA2A had a high sensitivity and specificity with an area under the curve value of 0.793 in differentiating AML patients from healthy individuals. THP-1 and HL-60 cells stably overexpressing circTADA2A exhibited reduced cell proliferation, and sensitized cell to ferroptosis by a ferroptosis inducer RSL3. Moreover, circTADA2A could counteracted Ferrostatin-1-induced inhibition of ferroptosis. Mechanistically, circTADA2A act as a sponge for miR-638, and upregulation of miR-638 expression could restore cellular phenotypes induced by circTADA2A. Our findings demonstrated that circTADA2A suppresses cell proliferation and promotes ferroptosis by sponging miR-638 during AML progression.