Theranostics
·
2026
·Vol. 16
·期 7
·3408
·
IF 13.3
Binbin Ji,Hui Guo,Rong Xing,Miaomiao Sun,Yu Cheng,Chen Yao,Hanyong Zhu,Xuerong Wang,Ruihan Jiang,Xin Chen,Zimeng Liu,Suyan Wang,Fei Xu,Fangyu Zhang,Fuxing Dong,Xiucheng Pan,Jing Yang,Yuchen Pan
Rationale: Macrophage phagocytosis is essential for pathogen clearance during sepsis. We previously demonstrated that the glycolytic enzyme 6-phosphofructokinase, muscle type (PFKM), modulates macrophage functions and its deficiency alleviates sepsis in mice. However, the function of PFKM in regulating macrophage phagocytosis remains unclear. Methods: CD14 + monocytes were sorted by flow cytometry from healthy volunteers and septic patients, and the subcellular localization of PFKM was assessed by immunofluorescence. Nuclear translocation mechanisms and PFKM-p53 interaction were identified by Co-immunoprecipitation coupled with mass spectrometry (Co-IP/MS) and validated by Co-IP. Transcriptomic sequencing was used to identify the downstream target of the PFKM-p53 complex. Inflammatory cytokine levels were detected by ELISA and real-time RT-PCR, and the phagocytosis of macrophages was assessed by flow cytometry. Dual-luciferase reporter assays and ChIP were employed to investigate whether PFKM acts as a co-regulator of p53 in mediating Pdcd1 transcription. Nanobodies targeting PFKM-p53 were screened and subsequently synthesized according to the sequences. The effect of nuclear PFKM and the therapeutic effect of nanobodies were evaluated on the well-established sepsis mouse models induced by Escherichia coli or cecal ligation and puncture. Results: PFKM translocated to the macrophage nucleus during sepsis. Nuclear accumulation of PFKM impaired phagocytosis through a non-glycolytic “moonlighting” function and exacerbated sepsis. Mechanistically, PFKM interacts with p53, which facilitates its nuclear translocation. Subsequently, PFKM promotes p53 acetylation at K120, enhancing p53 binding to the Pdcd1 promoter and driving its transcription, thereby suppressing macrophage phagocytosis. Blocking the PFKM-p53 interaction with a nanobody, Nb07, restored phagocytosis of macrophages and alleviated sepsis in mice. Conclusion: Our data reveal the PFKM-p53-PD-1 axis that suppresses macrophage phagocytosis in sepsis and highlight the therapeutic potential of targeting this pathway with nanobody-based strategies.
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PubMed
高影响因子
BREAST CANCER RESEARCH
·
2026
·
IF 6.2
Jianwen Wang,Longdi Yao,Yuming Chen,Qiang Zhu,Wenjing Xu,An Xu,Xiang Li,Zhi Chen,Min Cheng,Qi Leng,Chunlian Li,Xintao Deng,Deyuan Fu
Background: Breast cancer (BC) is one of the most common malignant tumors in women worldwide, with metastasis and recurrence constituting major therapeutic challenges. F-Box Protein 5(FBXO5), a core component of the E3 ubiquitin ligase complex, is overexpressed in multiple cancers, but its specific role and underlying mechanism in BC remain unclear and require further investigation. Methods: We screened E3 ubiquitin ligase-related cancer dependency genes by integrating data from the DepMap and UniProt databases, and further analyzed transcriptomic data from TCGA and GEO to identify the candidate gene FBXO5. We then validated its expression at the mRNA and protein levels in BC tissues from two independent centers and in cell lines, employing qRT-PCR, Western blot, and immunohistochemistry.The effects of FBXO5 on the proliferation, migration, and invasion abilities of BC cells were evaluated in vitro and in vivo through colony formation, CCK8, Transwell assays, wound healing assays, IHC, and subcutaneous tumor formation in nude mice. The interactions between FBXO5 and proteins, as well as its main functions and pathways, were investigated using Co-IP, mass spectrometry, immunofluorescence confocal microscopy, molecular docking and bioinformatics analysis, with results validated by rescue experiments. Results: The mRNA and protein expression levels of FBXO5 were significantly upregulated in BC tissues from two centers, high expression of FBXO5 was significantly correlated with adverse clinicopathological features, including larger tumor size, positive nodal status, and elevated Ki-67, and was associated with poor overall survival (OS) and disease-free survival (DFS).In vitro and in vivo experiments confirmed that FBXO5 significantly affected the proliferation, invasion, and migration abilities of BC cells. Mechanistically, Co-IP, mass spectrometry, molecular docking and immunofluorescence confocal microscopy experiments confirmed that FBXO5 directly interacts with ribosomal protein L23a (RPL23A) and promotes its polyubiquitination at the K48 chain, thereby regulating its degradation. Further experiments showed that the ubiquitination-mediated degradation of RPL23A led to a decrease in the stability of Tumor Protein p53(p53) and facilitated its degradation by Proto-Oncogene MDM2 (MDM2). Conclusion: Our study establishes the existence of a novel FBXO5/RPL23A/MDM2/p53 oncogenic axis in BC. These findings thereby nominate FBXO5 as a promising therapeutic target for BC intervention.
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PubMed
高影响因子
Theranostics
·
2025
·Vol. 15
·期 6
·2360
·
IF 13.3
Bin Li,Jingyi Liu,Wanyu He,Yanlin Zhou,Man Zhao,Cong Xia,Xiaoyue Pan,Zhihua Ji,Ruoyu Duan,Hui Lian,Kai Xu,Guoying Yu,Lan Wang
Rationale: Acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) is a critical syndrome with a mortality rate of up to 40%, and it is characterized by a prominent inflammatory cascade. The inflammasome and pyroptosis play crucial regulatory roles in regulating various inflammatory-related diseases by serving as pivotal signaling platforms for inflammatory responses and mediating the release of substantial quantities of inflammatory factors. Our previous studies confirmed that GC-1, a clinical-stage thyroid hormone analog, effectively mitigated pulmonary fibrosis by restoring mitochondrial function in epithelial cells. However, the potential effects of GC-1 on macrophage inflammasome assembly and pyroptosis in lung injury as well as the underlying mechanisms, remain unclear. Methods: The effects of GC-1 on lung injury, oxidative damage and inflammation were evaluated in two murine models of ALI (LPS- or HCl-induced models) by assessing lung pathology, the concentrations of IL-1β and IL-18 in BAL fluid, inflammasome and the levels of inflammasome- and pyroptosis-related proteins. Additionally, the impact of GC-1 on ROS-mediated inflammasome assembly and pyroptosis was investigated by examining ROS levels, Nrf2 signaling, and inflammasome adaptor protein ASC levels in mouse alveolar macrophages and human THP-1 macrophages treated with LPS and ATP. The Nrf2 inhibitor ML385 and the mitochondrial-ROS inhibitor Mito-TEMPO were used to further elucidate the effect of GC-1 on the Nrf2-p53-ASC pathway. Results: GC-1 significantly alleviated inflammation and lung injury in ALI model mice, as indicated by pulmonary pathology, inflammatory cytokine levels, ROS production and pyroptosis rates. Consistently, GC-1 inhibited ASC recruitment and oligomerization in macrophages, which suppressed the gasdermin D-mediated release of IL-1β and IL-18. These findings indicated a reduction in inflammasome assembly and pyroptosis initiation. Further research revealed that GC-1 may mitigate oxidative stress induced by mitochondrial damage through Nrf2 signaling, thereby inhibiting the expression of ROS-activated p53 and the target gene ASC. This protective effect of GC-1 could be reversed by ML385 and mimicked by Mito-TEMPO. Conclusions: This study presents a novel mechanism for treating ALI in which GC-1 inhibits macrophage ROS-mediated inflammasome assembly and pyroptosis through Nrf2-p53-ASC pathway. These findings highlight the promising potential of the use of GC-1 as an anti-inflammatory and antioxidant drug in the treatment of ALI/ARDS.
研究领域:
药理学
·
免疫学
·
呼吸医学
·
炎症研究
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PubMed
高影响因子
Biomed Research International
·
2020
·
IF 2.276
Kiren Mustafa,Hassan Mohamed,Aabid Manzoor Shah,Shaoxuan Yu,Muhammad Akhlaq,Haifang Xiao,Shaoqi Li,Tahira Naz,Shaista Nosheen,Xueyuan Bai,Yuanda Song
Human liver cancer has emerged as a serious health concern in the world, associated with poorly available therapies. The Berberis genus contains vital medicinal plants with miraculous healing properties and a wide range of bioactivities. In this study, different crude extracts of B. lycium Royle were prepared and screened against Human Hepatocarcinoma (HepG2) cell lines. The water/ethanolic extract of B. lycium Royle (BLE) exhibited significant antiproliferative activity against the HepG2 cancer cell line with an IC50 value of 47 μg/mL. The extract decreased the clonogenic potential of HepG2 cells in a dose-dependent manner. It induced apoptotic cell death in HepG2 cells that were confirmed by cytometric analysis and microscopic examination of cellular morphology through DAPI-stained cells. Biochemical evidence of apoptosis came from elevating the intracellular ROS level that was accompanied by the loss of mitochondrial membrane potential. The mechanism of apoptosis was further confirmed by gene expression analysis using RT-qPCR that revealed the decline in Bcl-2 independent of p53 mRNA and a rise in CDK1 while downregulating CDK5, CDK9, and CDK10 mRNA levels at 48 h of BLE treatment. The most active fraction was subjected to HPLC which indicated the presence of berberine (48 μg/mL) and benzoic acid (15.8 μg/mL) as major compounds in BLE and a trace amount of luteolin, rutin, and gallic acid. Our study highlighted the importance of the most active BLE extract as an excellent source of nutraceuticals against Human Hepatocarcinoma that can serve as an herbal natural cure against liver cancer.
研究领域:
药理学
·
肿瘤学
·
植物学
·
分子生物学
·
遗传学
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PubMed
高影响因子
Toxicology Research
·
2015
·Vol. 5
·期 1
·160-167
·
IF 3.983
Liye Zhu,Tao Yu,Xiaozhe Qi,Bo Yang,Lei Shi,Haoshu Luo,Xiaoyun He,Kunlun Huang,Wentao Xu
OTA can induce hepatotoxicity. Our previous research has shown that miRNAs play important roles in the OTA-induced hepatotoxicity. And miR-122 is the most abundant miRNA in the liver and is involved in diverse biological processes. This study was performed to clarify the role of miR-122 in OTA-induced hepatotoxicity. The expression levels of miR-122 and the target genes were quantified by real-time PCR. The OTA-induced apoptosis of hepatocyte and HepG2 cells was evaluated using a TUNEL kit, a CCK-8 kit, a flow cytometer and Hoechst 33342. miR-122 was inhibited in HepG2 cells. The results revealed that OTA affected rat hepatocyte apoptosis. miR-122 decreased at 4 weeks but increased at 13 weeks in the OTA-treated livers, and increased in the OTA-treated HepG2 cells; and the mRNA levels of CCNG1 and Bcl-w increased at 4 weeks and decreased at 13 weeks in the high-dose OTA-treatment groups and decreased in HepG2 cells. The apoptosis of HepG2 cells displayed a dose-related increase with OTA. However, the inhibition of miR-122 greatly reduced OTA-induced apoptosis. p53 decreased in vivo and in vitro. miR-122 is a primary effector of OTA-induced hepatocyte apoptosis through the CCNG1/p53 pathway and Bcl-w/caspase-3 pathway in vivo and in vitro. And miR-122 plays an important role in OTA-induced hepatotoxicity.
研究领域:
分子生物学
·
毒理学
·
细胞生物学
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PubMed
高影响因子
JOURNAL OF MATERIALS CHEMISTRY
·
2011
·Vol. 21
·期 11
·3585-3596
·
IF 5.101
Fu-Wei Huang,Jin Yang,Jun Feng,Ren-Xi Zhuo,Xian-Zheng Zhang
In this paper, a highly efficient gene transfer vector with hepatocyte-targeted function, galactosylated poly(L-succinimide)-g-polyethylenimine-g-lactobionic acid (PSI-g-PEI-g-LA), was synthesized by conjugating lactobionic acid (LA) to biodegradable cationic PSI-g-PEI polymers. The physicochemical properties of PSI-g-PEI-g-LA including buffer capability, plasmid DNA (pDNA) binding ability, cytotoxicity, zeta potential and complex size were explored. Dynamic light scattering (DLS) reveals that PSI-g-PEI-g-LA can compactly condense pDNA into nano-sized particles with a hydrodynamic diameter of 95–175 nm. PSI-g-PEI-g-LA exhibits much higher cell-biocompatibility compared to PSI-g-PEI and PEI. The hepatocyte-targeted function was demonstrated by comparative studies on three polymeric vectors including PSI-g-PEI-g-LA, PSI-g-PEI and PEI25k. The transfection efficiency was evaluated in two different cell lines including asialoglyco protein receptor (ASGP-R) bearing HepG2 cells and ASGP-R-lacking HeLa cells. The in vitro transfection tests using different reporter genes indicate that PSI-g-PEI-g-LA displays higher transfection activity in HepG2 cells due to the specific interaction between LA segments and its ASGP-R on HepG2 cells compared with PSI-g-PEI and PEI. In contrast, the three polymers exhibit similar transfection activity in HeLa cells. PSI-g-PEI-g-LA displays better serum-tolerant transfection ability. Based on these analyses, PSI-g-PEI-g-LA was used as the vector to mediate the transfection of a tumor-suppressor gene p53. By means of western blotting analysis, flow cytometry and confocal laser scanning microscopy techniques, considerably high p53 gene expression and consequently strong p53-inducing apoptosis of HepG2 cells are distinctly observed.
研究领域:
生物技术
·
分子生物学
·
药学
·
癌症研究
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高影响因子
Scientific Reports
·
2025
·Vol. 15
·期 1
·1-17
·
IF 3.9
Xing Yamin,Li Guangyuan,Li Ganggang,Xu Jixuan,Zhang Ting,Li Mengxue,Gao Chunxiao,Fu Miaoran,Zheng Pengyuan,Chu Xiufeng
Metallothioneins (MTs) are a class of cysteine-rich proteins that actively participate in the cellular defense against free radicals. However, owing to the high heterogeneity among different MTs, comprehensive investigations are needed to determine the biological activities and distribution patterns of each MT in different tissues. In the present study, ectopic expression of MT1H significantly inhibited the proliferation of gastric cancer cells. Mechanistically, MT1H was transported into the nucleus and regulated the expression of key genes involved in nutrient transportation and homeostasis, such as SLC6A19, TTC39B, and ADM2, and thereby activating the p53 and autophagy pathways. Additionally, survival analysis of data from the TCGA and other databases revealed that gastric cancer patients with high expression of MT1H had longer survival. Furthermore, MT1H was undetectable in most gastric cell lines, but its expression was increased upon treatment with dexamethasone (Dexa) and the metal ion zinc. Therefore, MT1H emerges as a valuable tumor suppressor, a biomarker for the prognosis, and a promising therapeutic target in gastric cancer patients.
研究领域:
肿瘤学
·
分子生物学
·
细胞生物学
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PubMed
高影响因子
CELLULAR AND MOLECULAR LIFE SCIENCES
·
2026
·
IF 6.5
Yang Da,Shi Yaxing,He Jieqian,Wang Minyu,Wu Si,Yu Miao,Wang Shuang,Ji Shuai,Li Jiaxing,Gong Zheng,Ju Lincheng
Clear cell renal cell carcinoma (ccRCC) is the most common malignant cancer in kidney. Exploring new therapeutic targets is crucial to improving the ccRCC patients’ overall survival. Kinesin family member 15 (KIF15), a member of the kinesin superfamily proteins, participates in various biological processes. However, the role and action mechanisms of KIF15 in ccRCC remain unknown. We observed that KIF15 was highly expressed in ccRCC tissues and its expression was negatively correlated with the prognosis of ccRCC patients. In vitro experiments showed that KIF15 promoted the proliferation of ccRCC cells. Consistently, in vivo assays showed that KIF15 accelerated tumor growth in ccRCC-bearing nude mice. Further investigation revealed that zinc finger protein 750 (ZNF750) repressed the transcription of KIF15 by directly binding its promoter region, simultaneously repressing ccRCC cell growth. Mechanistically, KIF15 stabilized MDM2 via enhancing USP15-mediated deubiquitylation, thus suppressing the p53 pathway. Our study indicates the oncogenic role and molecular mechanisms of KIF15 in ccRCC progression and provides valuable insights into potential therapeutic strategies for ccRCC.
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高影响因子
STRUCTURE
·
2024
·
IF 4.4
Cheng Li,Yunqiang Bian,Yiting Tang,Lingyu Meng,Peipei Yin,Ye Hong,Jun Cheng,Yuchen Li,Jie Lin,Chao Tang,Chunlai Chen,Wenfei Li,Zhi Qi
Nucleic acid and protein co-condensates exhibit diverse morphologies crucial for fundamental cellular processes. Despite many previous studies that advanced our understanding of this topic, several interesting biophysical questions regarding the underlying molecular mechanisms remain. We investigated DNA and human transcription factor p53 co-condensates—a scenario where neither dsDNA nor the protein demonstrates phase-separation behavior individually. Through a combination of experimental assays and theoretical approaches, we elucidated: (1) the phase diagram of DNA-protein co-condensates at a certain observation time, identifying a phase transition between viscoelastic fluid and viscoelastic solid states, and a morphology transition from droplet-like to “pearl chain”-like co-condensates; (2) the growth dynamics of co-condensates. Droplet-like and “pearl chain”-like co-condensates share a common initial critical microscopic cluster size at the nanometer scale during the early stage of phase separation. These findings provide important insights into the biophysical mechanisms underlying multi-component phase separation within cellular environments.
研究领域:
生物物理学
·
分子生物学
·
细胞过程
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PubMed
高影响因子
Oxidative Medicine and Cellular Longevity
·
2020
·
IF 5.076
Marwan Almoiliqy,Jin Wen,Eskandar Qaed,Yuchao Sun,Mengqiao Lian,Haithm Mousa,Mahmoud Al-Azab,Mohamed Y. Zaky,Dapeng Chen,Li Wang,Abdulkarem AL-Sharabi,Zhihao Liu,Pengyuan Sun,Yuan Lin
The aim of this study was to characterize and reveal the protective effects of cinnamaldehyde (CA) against mesenteric ischemia-reperfusion- (I/R-) induced lung and liver injuries and the related mechanisms. Sprague-Dawley (SPD) rats were pretreated for three days with 10 or 40 mg/kg/d, ig of CA, and then induced with mesenteric ischemia for 1 h and reperfusion for 2 h. The results indicated that pretreatment with 10 or 40 mg/kg of CA attenuated morphological damage in both lung and liver tissues of mesenteric I/R-injured rats. CA pretreatment significantly restored the levels of aspartate transaminase (AST) and alanine transaminase (ALT) in mesenteric I/R-injured liver tissues, indicating the improvement of hepatic function. CA also significantly attenuated the inflammation via reducing myeloperoxidase (MOP) activity and downregulating the expression of inflammation-related proteins, including interleukin-6 (IL-6), interleukin-1β (IL-1β), cyclooxygenase-2 (Cox-2), and tumor necrosis factor receptor type-2 (TNFR-2) in both lung and liver tissues of mesenteric I/R-injured rats. Pretreatment with CA significantly downregulated nuclear factor kappa B- (NF-κB-) related protein expressions (NF-κB p65, NF-κB p50, I kappa B alpha (IK-α), and inhibitor of nuclear factor kappa-B kinase subunit beta (IKKβ)) in both lung and liver tissues of mesenteric I/R-injured rats. CA also significantly downregulated the protein expression of p53 family members, including caspase-3, caspase-9, Bax, and p53, and restored Bcl-2 in both lung and liver tissues of mesenteric I/R-injured rats. CA pretreatment significantly reduced TUNEL-apoptotic cells and significantly inhibited p53 and NF-κB p65 nuclear translocation in both lung and liver tissues of mesenteric I/R-injured rats. CA neither induced pulmonary and hepatic histological alterations nor affected the parameters of inflammation and apoptosis in sham rats. We conclude that CA alleviated mesenteric I/R-induced pulmonary and hepatic injuries via attenuating apoptosis and inflammation through inhibition of NF-κB and p53 pathways in rats, suggesting the potential role of CA in remote organ ischemic injury protection.
研究领域:
药理学
·
病理生理学
·
免疫学
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PubMed
高影响因子
Cancer Management and Research
·
2020
·
IF 2.886
Mei Zeng,Yuhua Chen,Xintao Jia,Yan Liu
Background Epstein-Barr virus (EBV) has been indicated in the development of some tumors, including lymphoma. However, the potential role of latent membrane protein 1 (LMP1) encoded by EBV in the tumorigenesis of lymphoma remains debated. Herein, we examined the function of LMP1 in lymphoma. Methods The expression of LMP1 was downregulated or upregulated in EBV negative cell line SNT-8 and positive cell line KHYG-1, respectively. Subsequently, the cell viability, apoptosis, as well as the expression patterns of p53, mouse double minute 2 (MDM2), B-cell CLL/lymphoma 2 (Bcl-2) and NF-κB were evaluated. Next, the binding relationship between MDM2 and p53 along with p53 ubiquitination in cells was tested by Western blot and co-immunoprecipitation. Finally, the effects of LMP1 on lymphoma cell growth through p53, Bcl-2 and NF-κB pathways were verified by functional rescue experiments. Results Overexpression of LMP1 promoted KHYG-1 cell growth and inhibited cell apoptosis. Moreover, LMP1 upregulation significantly enhanced the activation of NF-κB pathway, thus increasing MDM2 binding to p53, leading to p53 ubiquitination and degradation as well as Bcl-2 expression enhancement. Further inhibition of the NF-κB pathway or Bcl-2 expression significantly weakened the promotive role of LMP1 in the growth of KHYG-1 cells. Conclusion EBV-LMP1 promoted the p53 ubiquitination and degradation by activating NF-κB signaling pathway and the following binding of MDM2 and p53 in cells to enhance Bcl-2 expression, thus promoting the growth of lymphoma cells and inhibiting cell apoptosis.
研究领域:
肿瘤学
·
分子生物学
·
细胞生物学
·
病毒学
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PubMed
高影响因子
JOURNAL OF CELLULAR PHYSIOLOGY
·
2018
·Vol. 234
·期 5
·7587-7599
·
IF 3.923
Junfa Yang,Yuchen Lu,Peipei Yang,Qingfeng Chen,Yang Wang,Qi Ding,Tao Xu,Xiaofeng Li,Changyao Li,Cheng Huang,Xiaoming Meng,Jun Li,Lei Zhang,Xiao Wang
Activation of quiescent hepatic stellate cells (HSCs) is the major event in liver fibrosis, along with enhancement of cell proliferation and overproduction of extracellular matrix. Recent findings suggest that senescence of activated HSCs might limit the development of liver fibrosis. The p53, a guardian of the genome is associated with liver fibrosis, has been shown to regulate HSCs senescence. In this study, we report that microRNA-145 (miR-145) and p53 were downregulated in vivo and in vitro, concomitant with the enhanced expression of zinc finger E-box binding homeobox 2 (ZEB2). In addition, overexpression of miR-145 and p53 led to upregulation of the number of senescence-associated β-galactosidase-positive HSCs and the expression of senescence markers p16 and p21, along with the reduced abundance of HSC activation markers α-smooth muscle actin and type I collagen in activated HSCs. Furthermore, silencing of ZEB2 promoted senescence of activated HSCs. Moreover, we also demonstrated that miR-145 specifically targeted the 3′-untranslated regions of ZEB2. In vitro promoter regulation studies show that ZEB2 could bind to the E-box of the p53 promoter as well as inhibit its promoter activity and thus suppress the expression of p53, which in turn repressed activated HSCs senescence. Taken together, our results describe a novel miR-145-ZEB2-p53 regulatory line might participate in the senescence of activated HSCs and might carry potential therapeutic targets for restraining liver fibrosis.
研究领域:
分子生物学
·
细胞生物学
·
肝病学
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PubMed
高影响因子
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