Human Cell最新文献

The objective of this research was to investigate the effects of proven antioxidant compounds identified in Nitraria retusa leaf extracts (Nr-extracts) on glioblastoma (GBM) cells. The antioxidant capacity was evaluated in nonenzymatic, enzymatic, and cellular systems. The antitumoral effect was demonstrated first by studying the inhibition of glioblastoma cell growth, then by examining the inhibition of cell adhesion to various purified extracellular matrix (ECM) proteins, and finally by assessing the ability of Nr-extracts to abolish the invasion of collagen gel by human glioblastoma cell spheroids. The methanolic extract (Nr-MeOH), through its isorhamnetin and carbohydrate derivatives, showed the highest antioxidant capacity. The cytotoxicity study of Nr-extracts, in the absence and presence of TEMODAL (TPZ), revealed that the chloroform extract (Nr-Chl), through its β-sitosterol and in combination with TEMODAL, exerted a more significant apoptotic effect on glioblastoma cells than TEMODAL alone. However, only the Nr-Chl and Nr-MeOH extracts inhibited the attachment of U87 cells to fibronectin, vitronectin, and collagen I. In addition, Nr-MeOH more significantly reduced U87 cell invasion compared with Nr-Chl extract. In conclusion, flavonols and sterols demonstrated strong antitumoral and free radical-scavenging activities, suggesting that a potentially synergistic therapeutic approach for glioma can be developed.

Being topmost cancer in India, oral cancer management warrants discovery of novel biomarkers, treatment strategies, and targets to help with early diagnosis, treatment, and recovery. To have a continuous supply of cells, the study was aimed at generation and characterization of established cell line from buccal mucosa (BM) tumors from patients of Indian origin which can be developed as a pre-clinical tool for biomedical application. Surgically resected tumor tissue from histo-pathologically confirmed oral cancer were processed for explant culture. TBM-02 cell line was passaged and characterized for morphology and function. Further, the cell line was silenced for inflammasome pathway gene NLRP3 to evaluate its linkage with oral cancer tumorigenesis. TBM-02, successfully established from BM, was maintained up to 100 passages, exhibited epithelioid morphology, high EpCam expression and triploid ploidy with chromosomal aberrations. Novelty and human origin of TBM-02 was authenticated by Short Tandem Repeats profiling and comparison with DSMZ database. TBM-02 revealed tumorigenic potential in vitro and in vivo which was abrogated on silencing NLRP3. Increased expression of NLRP3, hallmark of chronic inflammation in TBM-02, was validated at protein and gene level and in xenograft. TBM-02 demonstrated migratory potential and was found to be a sensitive tool to study drug response. RNA sequencing demonstrated upregulation of oral cancer-associated genes and pathways. Thus, in current study, we have reported development of novel cell line from early-stage buccal mucosa cancer patient which has a strong potential to be developed and to be used as pre-clinical model for improving oral cancer management and therapeutics.

Ovarian cancer is one of the most lethal gynecologic malignancies contributing to cancer-related deaths among women. However, the mechanisms underlying its development remain poorly understood. This study aimed to investigate the role of STARD7 in tumorigenesis and its potential molecular mechanisms in ovarian cancer. Cell proliferation was assessed using the Cell Count Kit-8 and colony formation assays. The migration and invasion capabilities of ovarian cancer cells were evaluated through wound healing and Transwell invasion assays. A xenograft tumor model was established in nude mice to examine the role of STARD7 in tumor growth in vivo. The results demonstrated that the knockdown of STARD7 in A2780 and SK-OV-3 cells inhibited cell proliferation, invasion, and migration, while overexpression of STARD7 yielded opposing results. Furthermore, STARD7 was found to enhance the levels of phosphorylated p65 proteins and nuclear protein p65. Treatment with the NF-κB inhibitor PDTC inhibited STARD7 overexpression-induced cell proliferation, invasion, and migration. These results suggest that the activation of the NF-κB pathway is involved in STARD7-mediated ovarian cancer progression. Consistent with the in vitro findings, similar results were observed in in vivo ovarian cancer xenograft models. Additionally, dual luciferase reporter assays indicated that STARD7 was regulated by RUNX1-IT1, which acted as a sponge for miR-377-3p. Collectively, these findings suggest that STARD7 may serve as an independent prognostic indicator, promoting ovarian cancer development by activating the NF-κB pathway. This research expands the current understanding of NF-κB regulatory networks and positions STARD7 as a promising therapeutic target for developing pathway-specific interventions.

Circular RNAs (circRNAs) are involved in cardiovascular disease development and progression. We identified has_circ_0000437 from three pairs of clinical samples using a circRNA microarray and predicted the has_circ_0000437 sponge let-7f-5p using bioinformatics. We expanded the sample to confirm the expression, correlation, and diagnostic value of has_circ_0000437, let-7f-5p and RAS-like proto-oncogene B (RALB) in rheumatic valvular heart disease (RVHD). The effects of has_circ_0000437 and let-7f-5p on biological function were observed in hVICs cells. To explore the molecular pathogenesis of has_circ_0000437 in the RVHD process, the target binding protein RAS-like proto-oncogene B (RALB) of has_circ_0000437 was obtained using label-free mass spectrometry and parallel reaction monitoring. The influence of the has_circ_0000437/let-7f-5p/RALB axis on the expression of mitogen-activated protein kinase (MAPK)-related proteins in the inflammatory signaling pathway predicted by Kyoto Encyclopedia of Genes and Genomes (KEGG) during RVHD was measured using Western blotting. We found that let-7f-5p was at low expression and negatively correlated with has_circ_0000437, which had a diagnostic value with an area under the curve of 0.998 after the combined diagnosis. Has_circ_0000437 and let-7f-5p had binding sites, and let-7f-5p alleviated the effect of has_circ_0000437 on the proliferation, migration, and cycle progression of hVICs. The inhibitory effect of has_circ_0000437 on hVICs apoptosis was diminished. Mechanism studies showed that has_circ_0000437 promotes the MAPK pathway through has_circ_0000437/let-7f-5p/RALB axis. These findings suggest that the has_circ_0000437/let-7f-5p/RALB axis promotes the MAPK pathway, proliferation, migration, and cycle progression in RVHD, inhibiting the apoptosis process, thereby promoting RVHD development. This pathway may suggest a target for RVHD diagnosis and treatment.

Currently, the ex vivo expansion of hematopoietic stem/progenitor cells (HS/PCs) is one of the effective ways to address the shortage of HS/PCs for clinical transplantation. Therefore, the optimal in vitro expansion strategy for HS/PCs is a critical issue to be addressed in current researches. In this study, we separated UCB Mononuclear cells (MNCs) and CD34⁺ cells, and cultured them for 14 days under serum-free conditions (only growth factors added). Then, the effects of the growth factors on HS/PCs expansion were analyzed and compared. The results showed that under short-term culture (< 7 days) the number of nucleated cells expanded significantly more in CD34⁺ cells than in MNCs, and the content of CD34⁺ cells was also significantly greater than that of CD45⁺CD34⁺ and CD34⁺CD38^- cell subpopulations in MNCs. It is confirmed that static suspension culture without serum was unable to promote the expansion of HS/PCs for a long term (14 days) and maintain their primitive stem cell activities. Under identical culture conditions, cell products harvested by expansion with CD34⁺ cells as the starting cell source outperformed MNCs in terms of hematopoietic stem/progenitor cell yields and hematopoietic reconstitution capacity.

Pathological cardiac hypertrophy develops as a maladaptive response to sustained pressure overload, transitioning from compensatory adaptation to dysfunction. Understanding its molecular mechanisms is crucial for developing therapeutic strategies. Here, we identified ovarian tumor (OTU) domain-containing ubiquitin aldehyde-binding protein 2 (OTUB2) as a key regulator of pathological cardiac hypertrophy. OTUB2 expression was significantly upregulated at both transcriptional and translational levels in transverse aortic constriction (TAC)-induced hypertrophic hearts and phenylephrine (PE)-stimulated cardiomyocytes. In vivo, cardiomyocyte-specific OTUB2 overexpression via AAV9 exacerbated TAC-induced cardiac remodeling, manifested by increased heart weight/body weight ratio, impaired ejection fraction, ventricular dilatation, and enhanced fibrosis (as shown by Picrosirius red staining). In neonatal rat cardiomyocytes (NRCMs), OTUB2 overexpression aggravated while its knockdown attenuated PE-induced cardiomyocytes hypertrophy. Mechanistically, OTUB2 upregulated both total and GTP-bound active Rac1, thereby activating the downstream MEK/ERK pathway. Notably, pharmacological inhibition of Rac1 activation with NSC23766 abolished OTUB2-mediated hypertrophic responses in PE-treated cardiomyocytes. Our findings establish the OTUB2/Rac1 axis as a novel regulator of pathological cardiac hypertrophy and a potential therapeutic target for cardiac remodeling.

We aimed to generate immortalized stromal cell lines from the ovarian and fallopian tube tissues of a single patient using Sendai virus (SeV) vectors and identify candidate stromal genes involved in ovarian carcinogenesis. Tissues were collected from a 48-year-old woman with endometrioid borderline tumors and endometriomas. Primary cultures were established from the right ovarian endometrioma, left ovarian surface, bilateral fallopian tube, and endometrial surface. Immortalization was achieved using SeV vectors encoding human telomerase reverse transcriptase (TERT), B lymphoma Mo-MLV insertion region 1 homolog (Bmi-1), and Simian virus 40 large T antigen (SV40T). Morphologically, the established cells exhibited spindle-shaped fibroblast-like features and expressed stromal markers (Vimentin-positive, Keratin-negative), confirming their stromal origin. Genetic and molecular changes associated with immortalization were evaluated via chromosomal analyses, transcriptome sequencing, and reverse transcription-polymerase chain reaction (RT-PCR). SeV-infected stromal cell lines retained their proliferative capacity for over 25 passages, whereas non-infected primary cells lost their epithelial characteristics and underwent senescence after five passages. Chromosomal abnormalities were more prevalent in stromal cells derived from the ovarian endometriomas, suggesting early genomic instability. Transcriptomic profiling and RT-PCR revealed upregulation of matrix metallopeptidase 1 (MMP1), pregnancy-associated plasma protein A (PAPPA), and C-X-C motif chemokine ligand 1 in cyst-derived stromal cells compared to those from the normal ovary and fallopian tube, implicating these genes in extracellular matrix remodeling and tumor-stroma crosstalk. We established immortalized ovarian and fallopian tube stromal cell lines using SeV-based vectors. The cyst-derived stromal cells exhibited early chromosomal instability and overexpression of MMP1 and PAPPA, supporting their potential role in ovarian carcinogenesis. These immortalized stromal cell lines provide a novel and stable platform for mechanistic studies and may contribute to biomarker discovery and therapeutic target development in ovarian cancer.

Dead-box RNA helicases (DDXs) are a family of proteins with roles in RNA metabolism, regulating processes such as RNA splicing, translation, and ribosome assembly. Recently, their functions have expanded to include essential roles in autophagy-a cellular degradation pathway crucial for maintaining homeostasis-and oncogenesis, notably in glioblastoma. Glioblastoma is characterized by rapid proliferation, invasiveness, and resistance to conventional treatments, making it a formidable clinical challenge. Emerging evidence suggests that specific DDXs may influence multiple key pathways that contribute to gliomagenesis, the process of glioma formation including cell cycle regulation, epithelial-to-mesenchymal transition (EMT), angiogenesis, immune modulation, anti-inflammatory signaling, and autophagy. Understanding the dual role of DDXs in autophagy and gliomagenesis may reveal potential therapeutic targets, as manipulating these helicases could disrupt cancer cell adaptation mechanisms and slow tumor progression. We have also explored the potential of autophagy inhibitors to enhance the efficacy of current therapeutics. This review aims to explore the implications of DDXs in glioblastoma, focusing on their interactions with cellular pathways, and highlights the need for further investigation into how these proteins could be leveraged for therapeutic benefit.

Clear cell renal cell carcinoma (ccRCC) is the most common type of renal cell carcinoma, and exploration of its molecular mechanism benefits for developing more effective molecular targeted drugs. CDC6 has been found to be highly expressed in a variety of malignancies and plays oncogenic role; however, its function in ccRCC has not been elucidated. In this work, immunohistochemical (IHC) staining was used to detect protein expression of genes in clinical tissues. qPCR and WB were used for expression detection of mRNA and protein levels in cells. The Celigo assay, plate cloning assay, flow cytometry, and wound-healing/Transwell assays were used to detect cell proliferation, colony formation, apoptosis, and migration, respectively. A subcutaneous xenograft model in nude mice was used to verify the function of CDC6 in vivo. The results of clinical sample-related detection as well as analysis showed that CDC6 was highly expressed in ccRCC and was significantly associated with higher tumor malignancy as well as worse patients' prognosis. Knockdown of CDC6 in ccRCC cells significantly inhibited cell proliferation and migration while promoting apoptosis, and inhibited in vivo growth of transplanted tumors in animal models. Mechanistically, RRM2 is identified as a potential downstream effector molecule that has co-expression characteristics with CDC6 and whose expression levels are regulated by it. More importantly, RRM2 knockdown mediated tumor suppression could partially reversed CDC6 overexpression induced tumor promotion. This study identified CDC6/RRM2 axis as a potential target for development of novel targeted therapy for ccRCC treatment.