Human Cell最新文献

The glycosylphosphatidylinositol (GPI)-anchored serine protease prostasin has been reported to have increased expression with tumor-promoting properties in some cancer types, while expression is lost and prostasin displays tumor-suppressing properties in other cancer types. Due to these context-dependent and opposing expression patterns and functions of prostasin, characterization of each cancer type is important. In the present study, we aimed to determine the expression of prostasin in the normal cervix and in cervical squamous cell carcinoma (CSCC), the most common type of cervical cancer. We found that prostasin protein is expressed in both murine and human cervix and is consistently localized on the cell surface in suprabasal layers of squamous cells in healthy cervical epithelia. To assess whether prostasin protein is differentially expressed during cervical carcinogenesis, we performed a comprehensive immunohistochemical analysis of prostasin protein expression levels and localization in tissue arrays of paraffin-embedded human cervical carcinomas compared to the corresponding normal tissue. Prostasin protein is expressed in the well-differentiated cellular strata with expression patterns similar to pan-keratin and E-cadherin, and is lost during the dedifferentiation of epithelial cells, a hallmark of high-grade CSCC. The prostasin expression profile, with differential expression in cancer, provide valuable information that may give clues to the function(s) of this protease in normal epithelial biology and carcinogenesis.

Thyroid transcription factor-1 (TTF-1) is a lineage-specific marker for lung adenocarcinoma (LUAD), whereas the relatively minor subset of TTF-1-negative LUADs shows a poor prognosis and a limited response to therapy. However, its relationship with the tumor immune microenvironment remains poorly defined. How TTF-1 expression affects the immune context in LUAD was investigated, focusing on tumor-associated macrophages (TAMs) and T-cell infiltration. Immunohistochemical (IHC) analysis of 226 LUAD specimens showed that TTF-1-negative tumors were associated with epidermal growth factor receptor wild-type status, advanced stage, and worse progression-free and cancer-specific survivals. Notably, PD-L1 (programmed death-ligand 1) and PD-L2 expression in TAMs, but not in cancer cells, was significantly reduced in TTF-1-negative tumors. Public single-cell RNA sequencing data confirmed downregulation of CD274 (PD-L1) in TAMs from tumors with low expression of TTF-1-related genes. In contrast, PDCD1LG2 (PD-L2) expression showed less consistent patterns. On IHC analysis, infiltration of CD8⁺ and CD4⁺ T cells was modestly lower in TTF-1-negative tumors, accompanied by decreased HLA class I and II expressions. Transcriptomic analysis of The Cancer Genome Atlas LUAD cohort further showed lower interferon gamma (IFN-γ) signaling and decreased T cell-inflamed gene signatures in the low TTF-1-negative tumors. These findings suggest that TTF-1-negative LUAD exhibits more immune-suppressive features, with a relatively reduced antitumor immune response characterized by decreased T-cell infiltration and INF-γ signaling, which are related to PD-L1 and PD-L2 expressions in TAMs.

Lung adenocarcinoma (LUAD) continues to be a major contributor to cancer-related deaths due to its aggressive nature and resistance to current therapies, highlighting the need for novel molecular insights and therapeutic targets. This study investigated the function of exosomal lncRNA FGD5-AS1 in lung adenocarcinoma (LUAD) and its interaction with miR-1179 and CDH3. We discovered that FGD5-AS1 was substantially overexpressed in LUAD cells and exosomes under hypoxic conditions, while miR-1179, a tumor suppressor, directly targeted and downregulated CDH3. By sponging miR-1179, FGD5-AS1 serves as a competing endogenous RNA (ceRNA) to prevent the suppression of CDH3, thereby promoting LUAD cell growth, movement, and infiltration. It was demonstrated that knockdown of FGD5-AS1 or overexpression of miR-1179 significantly reduced tumor growth in vivo. These results demonstrate a novel exosome-mediated regulatory axis, suggesting that targeting the FGD5-AS1/miR-1179/CDH3 pathway could offer new therapeutic strategies for LUAD.

Cholangiocarcinoma (CCA) is a highly heterogeneous primary malignant tumor of the biliary tract. Intrahepatic and extrahepatic cholangiocytes originate from different sources, resulting in significant clinical, epidemiological, molecular, and genetic heterogeneity. Globally, the incidence and mortality rates of CCA are generally increasing, highlighting the need for more foundational research to support advances in clinical diagnosis and treatment. Tumor cell lines remain a crucial tool for unraveling the molecular mechanisms underlying tumor development and for the development of novel therapeutic strategies. The existing CCA cell lines are insufficient to meet research demands, underscoring the urgent need to establish new CCA cell lines. In this study, we successfully established a novel human CCA cell line, designated EBC-X1, derived from a Chinese patient. It exhibits robust proliferative capacity and was successfully passaged for more than 40 generations. STR analysis confirmed that EBC-X1 is a distinct human-derived CCA cell line. The population doubling time was 64.5 h. Karyotypic analysis revealed that EBC-X1 cells exhibit complex karyotypes, with 91% being sub-diploid and 9% being sub-triploid. The representative karyotype is 35, X, der(4), del(5)(q35), der(8), inv(9), der(11), rob(13;15). Upon inoculation into NXG mice, subcutaneous transplant tumors were efficiently formed. EBC-X1 is resistant to paclitaxel, fluorouracil, and oxaliplatin but sensitive to gemcitabine. This model holds significant potential for advancing our understanding of the biological characteristics and molecular mechanisms of distal CCA, as well as for facilitating drug development efforts.

The DNA repair enzyme 8-oxoguanine DNA glycosylase-1 (OGG1) plays a crucial role in the initiation of DNA base excision repair pathway by recognizing and excising the oxidative base lesions including 7,8-dihydro-8-oxoguanine (8-oxoG). Beyond its canonical function in DNA repair, OGG1 has been implicated in regulating inflammation-related genes, growth factor expression, and various cell death pathways, including apoptosis, parthanatos, and autophagy. These mechanisms are often involved in obstetric and gynecological disorders, which are frequently characterized by inflammation, endothelial dysfunction, and dysregulated cell death. As such, OGG1 emerges as a potential therapeutic target for these conditions. However, comprehensive reviews detailing OGG1's mechanistic roles in reproductive diseases remain scarce. This review aims to synthesize current knowledge primarily on non-canonical functions of OGG1, with a focus on its potential involvement in disorders such as endometriosis, polycystic ovary syndrome, uterine fibroids, and malignancies, and to highlight its promise as a therapeutic target.

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 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 (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 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 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.