Human Cell最新文献

The prognosis of gastric cancer with peritoneal dissemination is poor because of its resistance to chemotherapy. To investigate the mechanism of drug resistance in peritoneal metastasis, cancer organoids were established from the ascites of a patient with peritoneal metastases of gastric cancer. The histological characteristics of the tumors were preserved in the organoids. A co-culture system was established by overlaying human-derived mesothelial cells on gastric cancer organoids embedded in type IA collagen, mimicking peritoneal dissemination foci. When co-cultured with mesothelial cells, the proliferation of ascites-derived gastric cancer organoids and other primary gastric cancer organoids was suppressed. Soluble factors derived from mesothelial cells were involved in suppressing cell proliferation. Organoids in co-culture showed reduced sensitivity to paclitaxel. This co-culture model may provide a useful platform for studying drug resistance mechanisms in the microenvironment of gastric cancer peritoneal metastases.

The malignant progression of colorectal cancer (CRC) is closely related to cell stemness, but its regulatory mechanism has not been fully elucidated. This study found that ADAMTS14 was significantly highly expressed in CRC tissues and cell lines and was associated with poor prognosis in patients. Functional experiments have confirmed that ADAMTS14 enhances the stemness characteristics (such as upregulation of ALDH1A1, ALDH1A3, and CD133 expression) and spheroidization ability of CRC cells by activating the Wnt signaling pathway. Further mechanism studies have shown that the transcription factor ONECUT2 is also highly expressed in CRC and indicates a poor prognosis, and it can directly activate its transcription by binding to the ADAMTS14 promoter region. In conclusion, this study has revealed a novel mechanism by which the ONECUT2/ADAMTS14/Wnt axis regulates the stemness of CRC cells, providing a potential molecular target for targeted intervention.

The choroid plexus (ChP) is a key brain structure responsible for cerebrospinal fluid (CSF) production and forms a selective barrier that regulates brain homeostasis and immune surveillance. In vitro models of ChP are essential for studying CSF dynamics, viral entry, neuroinflammation, and CNS drug transport; yet current organoid protocols remain complex, slow, and difficult to reproduce. Here, we report a quick and robust method for the generation of human iPSC-derived ChP organoids that is xeno-free and serum-free, scalable, and reproducible. Early GSK3β inhibition and transient WNT modulation guide organoids toward cystic ChP-enriched structures, confirmed by ventricle-like morphology, and expression of canonical markers (TTR, ZO-1). This minimal workflow enables rapid production of ChP-like organoids that recapitulate ChP morphology and marker expression, providing a potential platform for studies of cerebrospinal fluid physiology, barrier modelling, and translational neuroscience.

TEX14 is essential for the formation and maintenance of the intercellular bridge (ICB) which is one of the major cellular junctions required for spermatogenesis. Previously, TEX14 was shown to block germ cell abscission prior to cytokinesis through the GPPX3Y (Gly-Pro-Pro-X-X-X-Tyr) motif. Although TEX14 could have the potential to inhibit cell proliferation, it is difficult for full-length TEX14 to be applied as anti-tumour agents because of its molecular size. In addition, the involvement of amino acids surrounding the GPPX3Y motif in the ICB formation as well as cell proliferation remains to be investigated for anti-tumour treatment. In this study, we show that partial peptides of TEX14, which include the GPPX3Y motif, are sufficient to inhibit the proliferation of a variety of cancer cells and efficiently induce apoptosis. In addition, the length and variation of amino acids surrounding the GPPX3Y motif might attenuate the efficiency of partial TEX14 peptides to inhibit cell proliferation as well as to induce apoptosis. Thus, our findings suggest that these TEX14 short peptides could be useful to suppress cell division in continuously proliferating cells such as cancer cells without affecting germ cell differentiation and have the potential as anti-tumour agents.

The lysyl oxidase family is a group of copper-containing amine oxidases involved in the remodeling of the extracellular matrix, and regulates cell signaling, metabolism, organ development, and immunity by modifying the cellular matrix. Among them, LOXL3 had been shown to regulate the proliferation, migration, and invasion of breast, colorectal, lung, melanoma, and gastric cancers. Differential expression of LOXL3 in tumor affects tumor progression by modulating immune microenvironment, where LOXL3 plays a role in regulating immune cell infiltration, immune checkpoints, and the establishment of tumor drug resistance. Here, we summarize the structure, mechanism, and functional role of LOXL3 in tumor and tumor immune microenvironment. Further, we explored strategies to improve the immunotherapy efficacy by targeting LOXL3 in tumor. Based on its properties in immunity, we explored strategies to improve the immune efficacy of LOXL3 in immunotherapy.

Relapsing and remitting mucosal inflammation can potentially spread throughout the colon in cases of ulcerative colitis (UC). N6-methyladenosine (m⁶A) is the most prevalent epigenetic change in RNA, and it is essential to numerous biological processes and diseases, including UC. This study aims to investigate the roles and mechanisms of Wilms tumor 1-associated protein (WTAP) and protein tyrosine phosphatase non-receptor type 1 (PTPN1) in the etiology of UC and as potential therapeutic targets. PTP1B (gene name PTPN1) was significantly upregulated in UC samples and lipopolysaccharide (LPS)-stimulated NCM460 cells, influencing cell viability, DNA synthesis, and apoptosis. WTAP was identified as an upregulated methyltransferase in UC and was found to enhance PTPN1 expression through m⁶A modification, thereby exacerbating LPS-induced inflammation and cellular dysfunction. The use of the m⁶A modification inhibitor Cycloleucine and PTPN1 knockdown attenuated these effects, highlighting the effect of WTAP-mediated m⁶A modification of PTPN1 on UC pathophysiology. Our findings indicate the significant role of the WTAP-mediated m⁶A modification of PTPN1 in exacerbating UC-related inflammation and cellular dysfunction. Targeting the WTAP-PTPN1 axis could provide a novel therapeutic approach for managing UC, thereby addressing the unmet need for effective treatments in patients who are unresponsive to current treatment regimens.

This study aims to screen for differentially expressed mRNAs (DEmRNAs) and lncRNAs (DELncRNAs) related to autophagy in bone nonunion through transcriptome sequencing, and explore their molecular mechanisms at the cellular level. In this study, 5 pairs of bone nonunion and bone healing tissue samples were collected for RNA sequencing to obtain DEmRNAs and DElncRNAs. The protein-protein interaction (PPI) network of autophagy-related differential genes was constructed. In addition, an lncRNAs-mRNAs interaction network was constructed. qRT-PCR was used to verify key autophagy-related genes. shBNIP3 was transfected into Wistar rat bone marrow mesenchymal stem cells (BMSCs), and 14 days after osteoinduction, cell migration and osteoblast differentiation were detected by transwell, alizarin red staining, and ALP staining. A total of 1108 DEmRNAs and 46 DElncRNAs were identified, and further screening resulted in 15 autophagy-related DEmRNAs. GO and KEGG analysis showed that autophagy-related DEmRNAs are mainly involved in autophagy-related pathways. In addition, the lncRNAs network included 356 relationship pairs, and 5 central autophagy-related mRNAs (BNIP3, DDIT3, SIRT2, PINK1, and BAG3) were identified. QRT-PCR verified that the expressions of BNIP3, SIRT2, PINK1, and BAG3 were up-regulated, while DDIT3 was down-regulated in patients with bone nonunion compared with patients with normal bone union. Besides, sh2BNIP3 inhibited BMSCs cell migration and osteogenic differentiation compared with Mock and BNIP3-NC groups in vitro. In this study, central autophagy-related DEmRNA may play an important role in bone nonunion, especially BNIP3, which will help us to further understand the pathogenesis of bone nonunion.

Mitochondrial dysfunction is a key contributor to septic cardiomyopathy, driving myocardial inflammation and apoptosis. This study found that Sushi-repeat containing protein X-linked 2 (Srpx2) is associated with mitochondrial damage and is downregulated in the myocytes of lipopolysaccharide (LPS)-treated rats. Sprague Dawley rats were intraperitoneally injected with LPS (10 mg/kg) to establish a septic cardiomyopathy model. Adeno-associated viruses (8.5 × 10¹¹ vg/mL) containing Srpx2-overexpressing plasmids were injected into rats through their tail vein. Srpx2 overexpression improved hemodynamics and decreased myocardial damage in LPS-treated rats. H9C2 cells were treated with LPS (10 μg/mL) to establish an in vitro septic model. The cells were then transfected with Srpx2-overexpressing plasmids. Srpx2 overexpression ameliorated mitochondrial damage, which was evidenced by restoring mitochondrial morphology, enhancing the complex activities, and elevating ATP and mitochondrial membrane potential levels in cardiomyocytes. Srpx2 overexpression reduced mitochondrial reactive oxygen species levels and superoxide generation in cardiomyocytes. Srpx2 overexpression decreased cleaved caspase-3 and 9 protein levels. Tumor necrosis factor-α and interleukin-1 beta levels were also reduced in cardiomyocytes with Srpx2 overexpression, suggesting reversal of inflammation. The RNA-sequencing data indicated that Srpx2 might regulate the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) pathway to protect cardiomyocytes. Srpx2 overexpression increased the p-PI3K and p-AKT protein levels. The treatment of H9C2 cells with 10 μM LY294002, a PI3K/AKT pathway inhibitor, reversed the protective effects of Srpx2 against mitochondrial damage and apoptosis. In conclusion, Srpx2 alleviates mitochondrial damage by activating the PI3K/AKT pathway, thereby reducing apoptosis and inflammation in septic cardiomyopathy.

Asthma is a chronic inflammatory disease characterized by airway hyperresponsiveness, airway remodeling, and persistent inflammation, with neutrophilic phenotypes often associated with severe and steroid-resistant cases. SerpinB1, a serine protease inhibitor, has been shown to modulate inflammatory processes through its interaction with neutrophil elastase (Elane); however, its specific role in asthma pathogenesis has not been fully clarified. In the present study, we investigated the function of SerpinB1 in an ovalbumin (OVA)-induced mouse model of asthma as well as in lipopolysaccharide (LPS)-stimulated BEAS-2B cells. Expression analysis revealed that SerpinB1 was downregulated in asthmatic mice. Overexpression of SerpinB1 markedly alleviated neutrophil-driven airway inflammation, reduced structural remodeling of the airways, and suppressed pyroptosis, as demonstrated by decreased expression of caspase-1, GSDMD, IL-1β, and NLRP3. Co-immunoprecipitation and immunofluorescence assays further confirmed that SerpinB1 directly interacts with Elane. Importantly, knockdown of Elane abolished the protective effects conferred by SerpinB1, indicating that the regulation of asthma-related pathology by SerpinB1 is mediated through Elane inhibition. In conclusion, SerpinB1 may mitigate airway inflammation and remodeling in asthma by targeting Elane and suppressing pyroptosis, supporting it as a potential therapeutic target for neutrophil-dominant asthma.

A 62-year-old woman with EGFR exon 19-mutant lung adenocarcinoma developed resistance to osimertinib after 13 months of treatment. At progression, genetic analysis revealed persistence of the EGFR mutation and acquisition of a TPM3-NTRK1 fusion. Cells from the pleural effusion at this stage were cultured and established as a continuous cell line, LUNK1. LUNK1 cells displayed epithelial characteristics, with a population doubling time of 57.66 h in vitro and 100% tumorigenicity in vivo. Xenograft histopathology showed poorly differentiated lung adenocarcinoma exhibiting solid, acinar, and micropapillary patterns. Immunohistochemistry confirmed positivity for CK7, TTF-1, Napsin A, EGFR, and pan-TRK. EGFR exon 19 deletion and TPM3-NTRK1 fusion were confirmed by whole-exome and RNA sequencing, respectively, and further validated by PCR and Sanger sequencing. Drug sensitivity assays revealed an IC₅₀ of 64.0 nM for entrectinib and 806.8 nM for osimertinib, indicating reduced sensitivity to EGFR inhibition. NTRK1 knockdown significantly increased osimertinib sensitivity (124.5-264.5 fold) and reduced entrectinib sensitivity (8.7-19.0 fold), suggesting NTRK1 as a key regulator of drug response. We provided experimental evidence for that TPM3-NTRK1 fusions can mediate acquired resistance to osimertinib in a new lung adenocarcinoma cell line. LUNK1 represents a useful preclinical model for investigating resistance mechanisms and assessing dual-targeted treatment strategies.