Human Cell最新文献

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.

Endothelin-1 (ET-1) plays a critical role in diabetic vasculopathy. Although clinical trials have shown promise for ET-1 receptor antagonists in treating diabetic nephropathy, their clinical use remains limited by adverse effects. MiR-454-3p targets ET-1. This study aimed to investigate the role of miR-454-3p in modulating ET-1 expression and related molecular changes in endothelial cells (ECs) under high glucose conditions using both bioinformatics and experimental approaches. Bioinformatics analysis identified 10 miR-454-3p target genes expressed in ECs previously implicated in diabetic vascular complications: ET-1, GJA1, IRF1, PIK3CB, TRPC3, SLMAP, ESR1, ITGB8, MAPK1, and PPARG. With the exception of PPARG, which protects ECs from hyperglycemia-induced damage, all have been reported to exacerbate endothelial dysfunction. Western blotting showed that high glucose increased ET-1 expression in human umbilical vein ECs (HUVECs) and human dermal microvascular endothelial cells (HDMECs), while miR-454-3p overexpression significantly suppressed this effect in both cell types. Conditioned medium (CM) from HUVECs transfected with miR-454-3p mimics enhanced eNOS expression in recipient cells, compared to control CM. Pre-treatment of HUVEC control CM with an anti-ET-1 antibody also increased eNOS expression, supporting that miR-454-3p promotes NOS production partly via ET-1 suppression. MiR-454-3p overexpression in HUVECs did not affect PPARG expression or cell proliferation. In conclusion, miR-454-3p overexpression inhibits high glucose-induced ET-1 expression in HUVECs and HDMECs, and promotes eNOS production without affecting PPARG expression in HUVECs. Our findings suggest that miR-454-3p modulates ET-1 expression under hyperglycemic conditions in vitro, which may provide a foundation for future studies exploring its potential application in managing diabetic vasculopathy.

Chronic plaque psoriasis (CPS) and lichen planus (LP) are two chronic, immune-mediated inflammatory skin conditions that often spontaneously heal, even if recurrence is common. In both diseases, the immune system mistakenly targets skin cells, leading to inflammation and visible cutaneous lesions. Although the pathogenesis of LP is not fully understood, it is thought to involve an alteration of epidermal self-antigens, which activates T cells lymphocytes causing a persistent chronic inflammation at the dermal-epidermal junction. In psoriasis (PSO), an involvement of not properly considered immune cells, such as dendritic cells, monocytes, macrophages, neutrophils, keratinocytes and natural killer (NK) cells is present. Trophoblast surface antigen 2 (Trop2) is a type I transmembrane glycoprotein belonging to the epithelial cell adhesion molecule (EpCAM) family involved in cell adhesion. We conducted a retrospective study on 30 patients with the following diagnoses: 10 cases of Lichen planus (LP), 10 cases of chronic plaque psoriasis (CPS) and 10 healthy skin samples (HS). In all these samples, we performed an immunohistochemical analysis of Trop2 expression. LP and CPS samples showed a significant decrease in Trop2 expression compared to HS. Contrarily to LP, basal and supra-basal layers of CPS samples were completely negative, while the medium layers were highly positive for Trop2. The sub-corneal layers were negative for Trop2 in both LP and CPS. Our findings suggest that Trop2 may play a protective or regulatory role in maintaining skin homeostasis, particularly in the setting of immune-mediated inflammation.

Multiple myeloma (MM) is a hematologic malignancy characterized by abnormal clonal plasma cells in the bone marrow. This study aims to investigate the mechanism by which the long non-coding RNA FEZF1 antisense RNA 1 (FEZF1-AS1) regulates ferroptosis in MM cells through KIAA1429-mediated N6-methyladenosine (m6A) modification, and to identify novel therapeutic targets for MM therapy. The expression levels of FEZF1-AS1, Vir-like m6A methyltransferase associated protein (KIAA1429, also known as also known as VIRMA), and OTU deubiquitinase, ubiquitin aldehyde-binding 1 (OTUB1) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analysis. Cellular viability, reactive oxygen species (ROS) accumulation, glutathione (GSH) levels, ferrous iron (Fe²⁺) concentration, malondialdehyde (MDA) content, and the protein levels of solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4), and acyl-CoA synthetase long-chain family member 4 (ACSL4) were assessed. The interaction between FEZF1-AS1 and insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3), as well as the interaction between IGF2BP3 and KIAA1429, was validated using RNA pull-down and RNA immunoprecipitation (RIP) assays. m6A and YTH N6-methyladenosine RNA-binding protein 1 (YTHDF1) on OTUB1 messenger RNA (mRNA) was analyzed. The stability of KIAA1429 mRNA and OTUB1 mRNA was also evaluated. In addition, the binding of OTUB1 to SLC7A11 and the ubiquitination status of SLC7A11 were determined by co-immunoprecipitation assays. The results showed that FEZF1-AS1, KIAA1429, and OTUB1 were highly expressed in MM cells. Knockdown of FEZF1-AS1 reduced cell viability and promoted ferroptosis. Mechanistically, FEZF1-AS1 bound to IGF2BP3, which enhanced the stability and expression of KIAA1429 mRNA. KIAA1429 facilitated m6A modification on OTUB1 mRNA, thereby promoting OTUB1 expression through YTHDF1. OTUB1 in turn stabilized SLC7A11 expression by deubiquitination. Overexpression of either KIAA1429 or OTUB1 partially reversed the pro-ferroptotic effect induced by FEZF1-AS1 inhibition in MM cells.

The occurrence of tumor metastasis is associated with the phenotype of tumor-associated macrophages (TAMs). Within the tumor microenvironment, TAMs undergo metabolic reprogramming. Aerobic glycolysis contributes to TAM polarization into an M2-like phenotype. Cedrol is a component extracted from herbal medicines. Anti-cancer activities of cedrol have been reported, however, its effect on the metabolic reprogramming and pro-metastatic ability of TAMs remains unclear. Here, we found that cedrol reduced THP-1-derived macrophage polarization into M2-like phenotypes. Cedrol impaired the promoting effects of TAMs on the migration and invasion of lung cancer cells. Tumor metastasis in C57BL/6 mice was reduced following cedrol treatment, accompanied by decreases in M2-like TAMs in the lung tissues. Cedrol treatment decreased glucose consumption and lactate production and downregulated glycolysis-associated gene expression. A glycolytic inhibitor (2-DG) was utilized to confirm that suppression of glycolysis in TAMs limited the metastasis of lung cancer cells. MYC was identified as a downregulated gene in cedrol-treated TAMs based on the mRNA sequencing analysis. MYC overexpression could reverse the effects of cedrol on TAMs. Cedrol treatment reduced MYC expression at least partly via the PI3K-Akt pathway. These findings suggest that cedrol exerts anti-tumor effects by inhibiting TAM polarization into an M2-like phenotype by modulating MYC-mediated aerobic glycolysis, indicating cedrol as a potential drug for lung cancer treatment.

Deoxycholic acid (DCA), a microbial-derived secondary bile acid, plays a multifunctional role in gastrointestinal (GI) carcinogenic through various molecular and cellular mechanisms. Mechanistically, DCA causes disruption of epithelial barrier function by occludin, downregulation of claudin-5, and disruption of ERK signaling, increasing permeability and inflammation. DCA initiates DNA damage by reactive oxygen species (ROS), production of hydroxyl radicals, and degradation of p53, triggering Poly (ADP-ribose) polymerase (PARP)-mediated DNA repair signals. DCA triggers pro-oncogenic signaling such as β-catenin, M3 muscarinic receptor (M3R) transactivation of Epidermal Growth Factor Receptor (EGFR), and Nuclear factor kappa B (NF-κB), promoting cell proliferation, synthesis of Mucin 2 (MUC2), and pro-inflammatory cytokine release (e.g., Interleukin-8 (IL-8), Interferon gamma (IFN-γ)). DCA also inhibits antitumor immunity by blocking Ca²⁺-Nuclear factor of activated T-cell (NFAT) 2 signaling in CD8⁺ T cells, thus disrupting cytotoxicity. DCA causes intestinal metaplasia and trans-differentiation in gastric and esophageal epithelial cells via KLF Transcription Factor 5 (KLF5)-caudal-related homeobox transcription factor 2 (CDX2) signaling. While acute levels of DCA induce apoptosis by mitochondrial membrane depolarization and caspase-9 activation, chronic accumulation leads to tumorigenesis through chronic inflammation, disruption of barrier function, and immune escape. DCA-heparin conjugates are antiangiogenic and chemo-sensitizing and offer new therapeutic windows. Taken together, these data provide evidence for the dualistic action of DCA and its central position as a microbial metabolite linking diet, barrier function, immunity, and GI carcinogenesis.

Histiocytic sarcoma is an extremely rare and aggressive malignant neoplasm characterized by immunophenotypic features of mature histiocytes. The mechanisms underlying its malignant transformation remain poorly understood; consequently, the development of effective therapies remains limited. Resected histiocytic sarcoma specimens were cultured using a modified air-liquid interface organoid method, serially passaged, and xenografted into NOD-scid IL2Rgnull mice. Tumors formed by xenografted organoids retained histological and genetic similarities with the original tumor. Genomic analysis revealed the activation of the Sonic Hedgehog signaling pathway and amplification of Yes-associated protein 1, a key effector of the Hippo pathway. Accordingly, we evaluated the sensitivity of the organoids to the Sonic Hedgehog inhibitor vismodegib and Yes-associated protein 1 inhibitor verteporfin, both of which demonstrated potent in vitro antitumor activity in organoid cultures. This model offers a valuable preclinical platform for investigating the molecular pathology of this rare malignancy and accelerating the development of targeted therapies.

This study analyzed diabetic nephropathy (DN)-related single-cell RNA sequencing (scRNA-seq) data from public databases and dissected the mechanism by which the sirtuin 1 (SIRT1)/autophagy-related 5 (ATG5) axis mediates high glucose (HG)-induced human renal glomerular endothelial cell (HRGEC) injury. The endothelium cluster was analyzed with DN-related scRNA-seq data (GSE131882 and GSE264268). HG-induced HRGEC injury was assessed by detecting cell viability, LDH release, apoptosis, EMT, and autophagy. SRT1720 was used to activate SIRT1 in cell models and STZ-induced mouse models. Renal dysfunction and pathological injury were assessed by detecting urinary albumin, serum creatinine, and BUN levels and performing histopathological staining (H&E, PAS, Masson, and TUNEL). Analysis of the endothelium cluster discovered that the autophagy pathway in the endothelial cluster was suppressed in early-stage DN patients and mice. Moreover, HG induced cell apoptosis and EMT in HRGECs, along with elevated acetylated levels of ATG5 and decreased protein levels of ATG5. SRT1720 decreased apoptosis, EMT, and elevated autophagic flux in HG-induced HRGECs, as well as improved renal function and histopathological changes, reduced EMT, and elevated autophagy in DN mouse models. However, Atg5 silencing reversed SRT1720-mediated alterations in these parameters. The SIRT1/ATG5 axis-dependent HRGEC autophagy restoration exerts a protective effect on the kidney during DN, offering a scientific ground for developing therapeutic strategies for DN based on autophagy regulation.

Ferroptosis, an iron-dependent type of regulated cell death driven by excessive lipid peroxidation, plays an important role in natural tumor suppression. In this study, we identified 23 ferroptosis-related genes associated with prognosis in kidney cancer datasets. Based on the expression profiles of these genes, we classified kidney cancer into four distinct subtypes and constructed a 9-gene risk score to predict the prognosis of patients. Our analysis revealed that patients classified into group III and those in the low-risk group demonstrated significantly better survival probability. Moreover, the risk score exhibited strong predictive accuracy for the prognosis of kidney renal clear-cell carcinoma (KIRC) patients. Among the identified genes, PEBP1 showed elevated expression in both subtype III and the low-risk group, suggesting that it may act as a critical tumor suppressor. To further evaluate this, we examined PEBP1 expression patterns and their clinical correlations using TCGA-KIRC and KIRP cohorts. The results indicated that PEBP1 deletion was strongly associated with poor prognosis, while reduced PEBP1 expression correlated with advanced disease progression in both KIRC and KIRP patients. Functional enrichment analysis suggested that PEBP1 may be involved in pathways related to fatty acid metabolism and oxidative phosphorylation. Experimental validation supported these findings, showing that PEBP1 overexpression suppressed the proliferation and migration of kidney cancer cells. Additionally, PEBP1 promoted the accumulation of lipid reactive oxygen species (ROS), an effect that was reversed by a ferroptosis inhibitor. Conversely, silencing PEBP1 counteracted the lipid ROS induced by RSL4, a ferroptosis activator. In summary, our results demonstrate that PEBP1 functions as a potential tumor suppressor in kidney cancer and may serve as a promising prognostic biomarker and therapeutic target.