Cell Biochemistry and Biophysics最新文献

Cervical cancer remains a frequently-occurring gynecologic health problem posing a great threat to women. Tuftelin1 (TUFT1), an acidic protein possessing secretory capacity, has been reported to drive cisplatin resistance in cervical cancer cells. Accordingly, the current study is intended to figure out the specific impacts of TUFT1 on the aggressive behaviors of cervical cancer cells and make an in-depth study into the related regulatory mechanism. Firstly, analysis of TUFT1 expression in cervical cancer cells was performed by RT-qPCR and Western blotting. Cervical cancer cell proliferation was estimated via CCK-8 and colony formation assays. Wound healing, transwell as well as sphere formation assays were used to appraise cell migration, invasion, and stemness, respectively. Western blotting examined the expressions of metastasis- and stemness-associated factors and RT-qPCR also tested the expressions of stemness-associated factors. Co-IP assay was used to verify the binding between TUFT1 and activating transcription factor 1 (ATF1). Subsequent ATF1 expression was examined by RT-qPCR and Western blotting after TUFT1 was silenced. After co-transfected with TUFT1 interference and ATF1 overexpression plasmids, aforementioned functional experiments were conducted again. Western blotting also analyzed the expressions of epidermal growth factor receptor (EGFR) signaling-associated proteins. The experimental data determined that TUFT1 expression was fortified in cervical cancer cells and TUFT1 absence diminished cervical cancer cell proliferation, migration, invasion, and stemness. Besides, TUFT1 bond to ATF1 and positively modulated ATF1 expression. Moreover, ATF1 elevation countervailed the impacts of TUFT1 insufficiency on the proliferation, migration, invasion, stemness as well as EGFR signaling in cervical cancer cells. Anyway, TUFT1 might interact with ATF1 to elicit pro-proliferation, pro-metastasis, and pro-stemness properties and inactivate EGFR signaling in cervical cancer, supporting that TUFT1 might be valued as a potential hallmark for cervical cancer.

Colorectal cancer (CRC) is the second leading cause of cancer-related mortality worldwide. While the incidence and mortality of CRC have decreased overall due to better screening, rates in adults under 50 have risen. CRC can manifest as inherited syndromes (10%), familial clustering (20%), or sporadic forms (70%). The gut microbiota, comprising mainly firmicutes and bacteroidetes, play a key role in CRC development and prevention. Indeed, CRC progression is influenced by the dynamic interaction between the gut microbiota, the intestinal barrier, the immune system, and the production of short-chain fatty acids. Not surprisingly, imbalance in the gut microbiota, termed dysbiosis, has been linked to CRC due to ensuing chronic inflammation, DNA damage, and oxidative stress. This may explain the notion that probiotics and fecal microbiota transplantation offer potential strategies for CRC prevention and treatment by restoring microbial balance and enhancing anti-cancer immune responses. This review appraises the roles of gut microbiota in promoting or preventing CRC. It also discusses the mechanistic interplay between microbiota composition, the intestinal barrier, and the immune system, with the hope of developing potential therapeutic strategies.

Breast cancer, the most common malignancy in females, and rhabdomyosarcoma (RMS), the most prevalent soft tissue sarcoma in children, remain significant clinical challenges. This study evaluated the anticancer potential and apoptotic signaling pathways of Gracilaria edulis extracts and identified their mechanisms of action against RMS and breast adenocarcinoma (MCF-7) cell lines. Cytotoxicity was assessed using MTT assays, while apoptotic potential was evaluated through phase contrast and fluorescence microscopy, caspase 3/7 activity, DNA fragmentation, and gene expression analysis of apoptosis regulatory genes. In silico analysis was also performed to examine the molecular interactions of bioactive compounds present in Gracilaria edulis with cancer-related proteins involved in apoptotic signaling. The methanol extract was fractionated into hexane, chloroform, and ethyl acetate, with the hexane fraction demonstrating the strongest cytotoxicity (IC₅₀RMS: 32.52 ± 2.15 μg/mL; IC₅₀MCF-7:29.84 ± 0.65 μg/mL) in MTT assays. Apoptotic features, including chromatin condensation, membrane blebbing, cellular shrinkage, and DNA fragmentation, were observed, particularly in RMS cells. The hexane fraction significantly activated caspase 3/7 in RMS cells, while lower activation was noted in MCF-7 cells, possibly due to the partial deletion of the CASP-3 gene. Real-time PCR analysis revealed differential gene expression, with p21 showing dominant upregulation in RMS cells and p53 being more prominently expressed in MCF-7 cells. These findings reflect their distinct roles in apoptotic signaling pathways. A significant increase in the Bax/Bcl-2 ratio in RMS cells (8.45) and MCF-7 cells (29.69) indicated a pro-apoptotic shift. GC-MS analysis identified key bioactive compounds, including 9-octadecenoic acid methyl ester, hexadecenoic acid methyl ester, and 1,2-benzenedicarboxylic acid mono(2-ethylhexyl) ester. In silico docking revealed that 1,2-benzenedicarboxylic acid mono(2-ethylhexyl) ester demonstrated the most promising binding interactions, particularly with BCL-2, while 9-octadecenoic acid methyl ester exhibited weaker binding affinities across all targets (p53, p21, and BCL-2), suggesting limited therapeutic relevance without structural optimization. However, the hexane fraction of G. edulis and its bioactive compounds remain promising as potential anticancer agents, warranting further in vitro and in vivo validation and molecular optimization.

Hepatocellular carcinoma (HCC) represents a significant global health challenge, characterized by a high incidence rate. Mitochondria have emerged as an important therapeutic target for HCC. Donafenib, a multi-receptor tyrosine kinase inhibitor, has been approved for the treatment of advanced HCC. However, the underlying mechanisms remain to be elucidated. In this study, we aim to investigate the effects of Donafenib on mitochondrial function in HCC cells. Firstly, we show that Donafenib induces mitochondrial oxidative stress in SNU-449 liver cancer cells by increasing mitochondrial ROS while reducing glutathione peroxidase (GPx) activity and the expression of Mn-SOD. We also demonstrate that Donafenib decreases mitochondrial membrane potential (MMP) and induces the opening of the mitochondrial permeability transition pore (mPTP). Furthermore, Donafenib reduces mitochondrial respiratory rate, COX IV activity, and ATP production. Notably, Donafenib induces mitochondrial fragmentation and reduces mitochondrial length by increasing the expression of DRP1, without affecting Mfn1 or Mfn2. Silencing of DRP1 protects against mitochondrial dysfunction induced by Donafenib, indicating that DRP1 plays a key role in mediating Donafenib's effects on mitochondrial function in HCC cells.

Acute myocardial infarction (AMI) is a critical cardiovascular disease with high disability and mortality rates, primarily caused by hypoxic injury to myocardial cells. This study investigates the role of the Vitamin D receptor (VDR) in cardiomyocytes under hypoxic conditions. VDR expression was characterized in human and hypoxic cardiomyocytes isolated from mice. To understand the downstream effects of VDR-related pathways, VDR was modulated using shRNA. RXR expression and localization were measured in hypoxic and sh-VDR cardiomyocytes. Oxidative stress and apoptosis levels were assessed and the effect of Vitamin D treatment was evaluated. VDR expression was found to be downregulated in the serum of AMI patients, similar to the hypoxic cardiomyocytes. Knockdown of VDR induced oxidative stress and apoptosis in normoxic cardiomyocytes, which could not be reversed by vitamin D treatment. Knock-down VDR in cardiomyocytes exposed to hypoxic induced apoptosis and reactive oxygen species via the HIF-1α/HO-1 axis. Overexpression VDR alleviated the expression levels of pro-inflammatory cytokines TNF-α, IL-6, and IL-1β. Our results indicated that VDR is crucial in reducing myocardial stress and apoptosis during hypoxic injury.

Scientific research is currently moving towards studies of natural antioxidants that could reduce the deleterious effects resulting from the toxicity of materials used in industry in order to preserve the environment and public health. Astaxanthin "king of antioxidants". is known for its remarkable protection against numerous environmentally harmful substances, but not against acrylonitrile, which is utilized in the manufacturing of plastics, rubber, and synthetic fibers. The goal of this investigation is to assess how astaxanthin can recover the normal CBC and normal organ architecture after acrylonitrile toxicity. Fifty rats were divided equally into five groups: positive control (acrylonitrile), negative control (saline), vehicle (corn oil), antioxidant (astaxanthin), and protective (astaxanthin + acrylonitrile). After sacrificing all the rats, CBC was done. Tissues of the heart, liver, and kidney were used for pathological examination and to assess TGF-β1 by immunohistochemistry, Our results showed an improvement in most of hematological parameters in the protective group compared to the acrylonitrile group. There was a significant improvement in RBC count, PCV, HB, RDW, and platelets, accompanied by a slight increase in MCV. Conversely, significant decreases were observed in PDW, MPV, MCH, and MCHC. Additionally, our results showed a slight decrease in WBCs and neutrophils, along with a slight increase in eosinophils and lymphocytes, and a significant increase in monocytes. There was a significant decrease in TGF-β1 level in rats treated with astaxanthin. All the organs showed an excellent recovery for the normal architecture. Astaxanthin can ameliorate acrylonitrile toxicity by restoring the normal levels of hematological parameters and histological structure.

The sea urchin (Paracentrotus lividus) shell investigation reveals a wealth of bioactive compounds. The bioactive ingredients were observed using UPLCMS/MS profiling. The anti-diabetic, antioxidant, antimicrobial, and anti-inflammatory qualities of P. lividus shell extract were assessed concerning NO, MDA, CAT, and SOD levels. Also, cytotoxic, and anti-angiogenic impact on colon (Caco-2) and breast (T47D) carcinoma cells and quantificated of Nrf2/HMOX-1 and HIF-1α/VEGF pathway expression were evaluated. Our findings indicate that the extract possesses remarkable antioxidant activity with IC₅₀ equal to (0.1056 ± 0.083 and 30.42 ± 1.52 μg/mL; for DPPH and ABTS⁺ respectively), antidiabetic with IC₅₀ (1.572 ± 0.13 μg/mL) and anti-inflammatory with IC₅₀ (2.090 ± 0.49 μg/mL). Notably, it exhibits potent anticancer effects against human breast (T47D) and colon (Caco-2) cancer cell lines, (30.55 ± 1.19 and 31.34 ± 1.22 µg/mL respectively). The extract induces oxidative stress and apoptosis, as evidenced by elevated NO and MDA levels, alongside reduced SOD and CAT activities. Moreover, the downregulation of Nrf2/HMOX-1 and HIF-1α/VEGF pathways expression suggests intricate molecular mechanisms underlying its anticancer properties, potentially involving the modulation of oxidative stress and angiogenesis. These findings underscore the sea urchin (P. lividus) shell as a potent reservoir of bioactive constituents with promising applications in pharmaceutical research and offering new avenues for drug discovery.

Vitamin A (VA) and its active form, retinoic acid (RA), are crucial for preserving hepatic stellate cells (HSCs) quiescence and reversing fibrosis. While C6orf120 is known to be involved in HSC activation, its role in RA signaling is unclear. This study found that C6orf120 knockdown markedly reduced CCL4-induced liver fibrosis and TGF-β1-induced activation in LX-2 cells, a human HSC line. This inhibition was associated with enhanced RA signaling, particularly affecting the RA receptor beta (RARβ). Inhibition of RARβ significantly reversed the protective effects of C6orf120 knockdown, indicating that RARβ signaling contributes to the inhibitory effect of C6orf120 knockdown on HSC activation. Our results reveal that C6orf120 inhibition could be a therapeutic strategy for liver fibrosis by regulating RARβ signaling.

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive solid tumor; however, the barrier of chemoresistance has yet to be overcome for longer survival. Aberrant gene expression due to epigenetic modification plays an important role in tumorigenesis and treatment. Super enhancers are epigenetic elements that promote targeted gene transcription and ultimately lead to chemoresistance. This study found that the expression of FOXM1 was higher in PDAC tissues and negatively correlated with prognosis. Through RNA sequencing and chromatin immunoprecipitation-sequencing analyses, FOXM1 was found to be regulated by a BRD4-associated super enhancer, which finally promoted gemcitabine resistance via TGFβ/Smad signaling pathway activation. Both TGFβ/Smad-specific inhibitor LY364947 and the BRD4 inhibitor JQ1 decreased the IC50 value of gemcitabine in vitro. Furthermore, combined gemcitabine and JQ1 therapy could not only enhance the therapeutic effect of gemcitabine but also reverse drug resistance in vivo. In conclusion, the super enhancer-associated gene FOMX1 contributes to gemcitabine resistance and is a promising target in PDAC treatment.

Pulmonary endothelial barrier dysfunction is a hallmark of sepsis-induced acute lung injury (ALI). Ophiopogonin D (OP-D), isolated from the roots of Ophiopogon japonicus, is involved in regulating inflammation, apoptosis and intestinal permeability. However, the role of OP-D in ALI has not been reported and the related mechanisms remain unclear. In this study, cecal ligation and puncture (CLP) was used to establish a septic ALI model in mice. We found that OP-D effectively alleviated lung pathological damage. Moreover, OP-D decreased pulmonary microvascular permeability, restrained the inflammatory response and apoptosis in murine lung tissues and LPS-exposed PMVECs. Specifically, OP-D exerted the beneficial effects via mediating the inactivation of HIF-1α-VEGF pathway, which was partly abrogated by the overexpression of HIF-1α. Collectively, our findings showed that OP-D protected against sepsis-induced ALI through improving pulmonary microvascular endothelial barrier dysfunction via suppressing HIF-1α-VEGF pathway.