Human Cell最新文献

miR-487a-3p shows the potential of modulating adipose-derived stem cells (ADSCs) differentiation. This study aimed to investigate the mechanism of miR-487a-3p on the osteogenic differentiation of ADSCs. In this work, ADSCs were induced to differentiate into osteoblasts. miR-487a-3p were regulated by miRNA mimics or inhibitors in ADSCs. Wnt family member 5A (WNT5A) siRNA was used to reverse miR-487a-3p inhibitor-induced effects on WNT5A expression in ADSCs. Fat mass and obesity-associated protein (FTO) in ADSCs were altered by shRNAs or overexpression vectors. Calcium nodule, ALP activity, and biomarkers of osteogenic differentiation (OD) were investigated. Rats received ovariectomy (OVX) to construct osteoporosis (OP) model, followed by ADSCs transplantation. Histopathological changes, bone histomorphometry, and detection of OD biomarkers were performed. We found that osteogenesis induced a decrease in miR-487a-3p expression and an increase in FTO expression. miR-487a-3p upregulation inhibited the OD of ADSCs, including decreases in calcium nodule formation, ALP activity, and OD biomarkers. miR-487a-3p downregulation showed the opposite role in OD. miR-487a-3p negatively regulated WNT5A in ADSCs. WNT5A silence reversed the effect of miR-487a-3p downregulation on OD. FTO silence caused the increase in m6A of pri-miR-487a. FTO overexpression inhibited DGCR8 recruitment in pri-miR-487a, and reversed the effect of miR-487a-3p upregulation in OD. ADSCs transplantation improved OP symptom in rats, including improvement of femur tissue, increase in percent bone volume and trabecular number, and upregulation in OD biomarkers. miR-487a-3p downregulation enhanced the therapeutic role of ADSC in rats with OP. Collectively, FTO regulated pri-miR-487a maturation via m6A-dependent pathway, which altered the WNT5A-mediated osteogenesis of ADSCs.

Intestinal failure-associated liver disease (IFALD) is a life-threatening complication of short bowel syndrome (SBS), characterized by cholestasis, hepatic steatosis, and hepatic fibrosis. Fish oil-based lipid emulsion (FO) has been demonstrated to ameliorate IFALD compared to soybean oil lipid emulsion (SO). However, the mechanisms underlying the beneficial effects of FO remain elusive. This study investigated the effects of FO on the activation of hepatic stellate cells (HSCs) that are primarily responsible for liver fibrosis by differentiating into fibroblasts in a transforming growth factor-β1 (TGF-β1) dependent manner. The human HSCs line LX-2 cells were stimulated with TGF-β1 in the presence of FO and SO. FO, but not SO, inhibited the degradation of lipid droplets induced by TGF-β1, suggesting that FO maintains HSCs in a quiescent state. Furthermore, FO suppressed LX-2 cell proliferation and partially abolished the autocrine regulation of TGF-β1 and subsequent activation of HSCs, as evidenced by the reduced expression of alpha-1 type I collagen (Col1a1) mRNA. These effects were specific to LX-2 and were not observed in the human hepatocellular carcinoma cell line HuH-7. The unique fatty acid composition of FO, characterized by high levels of long-chain polyunsaturated fatty acids with carbon chains of 20 or more, may contribute to its antifibrotic properties. These findings suggest that FO suppresses excessive HSCs activation while maintaining physiological functions, providing novel insights into the antifibrotic mechanisms of FO.

Regulatory T cell (Treg) immunome profile and targets in antiprogrammed cell death-1 (PD-1)/programmed cell death-ligand 1 (PD-L1) is a subject of extensive research, but there are still complexities in the area due to the nature of tumor microenvironment (TME). TME of solid tumors contains factors exerting a range of effects on Tregs including development, recruitment, expansion, stability and their immunosuppressive activity. Anti-PD-1 secondarily causes replenishment of intratumoral Tregs, which further intensify tumor immunosuppression. Besides, Treg depletion may also compromise immune checkpoint inhibitor (ICI) efficacy in addition to its other adversarial effects. Thus, a desired ICI booster is to use agents preferentially acting on intratumoral Tregs. Modulation of hypoxia, and regulation of Treg-related cytokines, chemokines, receptors and chromatin modifying factors in tumor TME provide supplementary approaches to anti-PD-(L)1. Factors acting on Tregs have diverse or even dual functions in TME. Treg expansion inhibitory and immunosuppressive tumor-associated macrophage (TAM) recruitment stimulatory effects of stimulator of interferon genes (STING), effector suppressor Treg activating and tumor-specific CD8⁺ T cell stimulatory effects of interleukin type 2 receptor alpha (IL-2Rα, also called CD25), cell type-dependent dual activities of CXCR3 and inducible T cell costimulatory (ICOS), exposure time-dependent dual effects of glucocorticoid-induced TNFR-related protein (GITR) and CD70 on Tregs and T cells, and exposure level-dependent dual activities of IFN-γ on Tregs are examples require consideration in designing Treg-based strategies. The main direction of this review is to provide updated information about targeting modulators of intratumoral Tregs with agents/compounds aiming to expand anti-PD-(L)1 efficacy and durability in solid tumors.

Nicotinamide N-methyltransferase (NNMT) is an S-adenosyl-l-methionine (SAM)-dependent cytosolic enzyme, and a growing body of evidence suggest that it plays an essential role in cancer progression. Recently, NNMT has a role in methylation metabolism and tumorigenesis and was associated with a poor prognosis against numerous cancers. In addition, it has been reported that NNMT has been overexpressed in the stroma of advanced high-grade serous carcinoma and may contribute to decreased survival. This study aimed to identify novel biomarkers to predict resistance and investigate their clinicopathologic significance in paclitaxel-resistant advanced or recurrent ovarian clear cell carcinoma (OCCC). Fluorescence-labeled two-dimensional differential gel electrophoresis (2D-DIGE), immunohistochemical, and MASCOT analyses allowed us to identify the cytoplasmic metabolic enzyme NNMT. In cultured cell studies, NNMT protein expression was higher in paclitaxel-resistant OVMANA and OVTOKO cells than in paclitaxel-sensitive KK and ES-2 cells. Furthermore, although analysis of clinical tissue samples showed no association with poor prognosis in 7 individuals with low NNMT expression in the cytoplasm of OCCC cells, high expression of NNMT in the cytoplasm of OCCC cells may be associated with low sensitivity to paclitaxel in OCCC and may have prognostic implications. Therefore, targeting therapy to reduce cytoplasmic NNMT expression levels may increase the sensitivity of OCCC to paclitaxel.

Most drug discovery studies use 2D cell cultures and animal models for screening new chemical entities (NCEs), which often leads to suboptimal results due to genetic variations, species differences, or lack of most physiological preclinical models. This is one of the most important reasons behind high rate of failure of drug candidate in the clinic, especially in oncology drug development projects. To address this issue, we developed a 3D pancreatic tumor spheroid model that better mimics the parental tumor architecture. We observed similar drug effects on cellular viability in both 2D cultures and 3D spheroids. However, cellular viability alone is insufficient to predict the translation of efficacy into clinical studies. A 3D multicellular tumor model is essential to comprehensively evaluate drug effects on the tumor microenvironment (TME), angiogenesis, and tumor biomarkers. Our model includes 3D monocellular and multicellular spheroids, which demonstrated a more relevant platform for potency evaluation. We used pancreatic ductal adenocarcinoma cells PANC-1 and PANC04.03 to conduct a comprehensive drug screening and assessed spheroid shrinkage and pre-vascularization. We also evaluated RT-qPCR analysis for gene expression of CSC markers (CD44, SOX2, KRT18), EMT markers (αSMA, vimentin) and the apoptotic marker (Annexin A1) under various conditions. Our findings highlighted the significant differences between 2D and 3D cultures, underscoring the importance of 3D multicellular models for predicting therapeutic markers and enabling comprehensive drug evaluation. In this study, MRTX1133 (a Phase I candidate of KRAS-G12D inhibitor) was used for testing our hypothesis. Treating the spheroids with MRTX1133 revealed enhanced drug response profiles compared to 2D cultures. This study underscores the critical importance of 3D multicellular model in preclinical drug screening and their potential to bridge the gap between in vitro studies and clinical outcomes.

The expression of collagen receptors by cancer cells serves a vital function in the regulation of cell behavior. These receptors are capable of sensing the signals generated by alterations in the collagen state, thereby contributing to the maintenance of cellular homeostasis. The discoidin domain receptor (DDR)1 functions as a critical sensor of collagen fiber state and composition, regulating tumor cell growth, response to therapy, and patient survival. We evaluated the role of collagen alpha-5(IV) chain (COL4A5) in nasopharyngeal carcinoma (NPC) and the detailed mechanism. GSE118719 and GSE68799 datasets were included to identify COL4A5 as a hub gene in NPC. Transcriptional activation of COL4A5 by nuclear factor 1/C (NFIC) mediated DDR1/Akt signaling activation, promoted NPC cell proliferation, migration, invasion, and curbed apoptosis in vitro, and exacerbated malignant progression of subcutaneous tumors in nude mice. NFIC and COL4A5 were significantly overexpressed in the tumors of NPC patients, and their expressions were significantly positively correlated. The overexpression of NFIC and COL4A5 was closely related to the tumor, node, metastases stage of NPC patients. Collectively, our results suggest that NFIC transcriptionally activates COL4A5 and upregulates its expression, which mediates DDR1/Akt signaling and promotes the malignant behavior of NPC cells, leading to NPC progression.

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.