We previously showed that proliferating cell nuclear antigen (PCNA) interacts with androgen receptor (AR) through a PIP-box (PIP-box4) at the N-terminus of AR and regulates AR activity. In this study, we further investigated PCNA/AR interaction. We identified a second PIP-box (PIP-box592) in the DNA binding domain of AR and found that dihydrotestosterone enhances the binding of full-length AR (AR-FL) but not a constitutively active variant (AR-V7) to PCNA. Treatment with R9-AR-PIP, a PIP-box4-mimicking small peptide, inhibits the PCNA/AR interaction, AR occupancy at the androgen response element (ARE) in PSA and p21 genes, and expression of AR target genes, and induces cytotoxicity in AR-positive castration-resistant prostate cancer (CRPC) cells. R9-AR-PIP also significantly inhibits transcriptional activity of AR-FL upon dihydrotestosterone stimulation and the constitutive activity of AR-V7. Moreover, R9-AR-PIP and PCNA-I1S, a small molecule PCNA inhibitor, inhibit the ARE occupancy by AR-FL and AR-Vs in CCNA2 gene that encodes cyclin A2 and cyclin A2 expression. Finally, we found that cyclin A2 is overexpressed in all CRPC cells examined, suggesting that it may contribute to the development of CRPC. These data indicate that targeting PCNA/AR interaction inhibits both AR-FL- and AR-Vs-mediated signaling and implicates it could be a novel therapeutic strategy against CRPC.

Sulforaphane (SFN), a lipophilic small-molecule compound, can be rapidly and completely absorbed upon entering the body. It has garnered extensive research attention for its potential as an antiaging, anticancer, antidiabetic, and antibacterial agent. However, its role and mechanisms of SFN on skeletal muscle postinjury regeneration have not been reported. This research demonstrated that SFN enhanced the regeneration after skeletal muscle injury and up-regulated the proliferation of mouse C2C12 myoblasts. RNA-transcriptome sequencing data revealed that SFN increased Prl2C2 transcription and JAK/STAT signaling pathway activity. CHIP and dual-luciferase reporter gene assays verified that STAT3 binds to the Prl2C2 promoter and regulates its transcription. Consequently, SFN influenced the JAK2/STAT3 signaling activity. Finally, the transcription of Prl2C2 and the proliferation of mouse C2C12 myoblasts were detected by adding JAK2 inhibitor and SFN. The results showed that the JAK2 inhibitor blocked the up-regulation of SFN on the transcription of Prl2C2 and the proliferation of mouse C2C12 myoblasts. The discovery of this phenomenon and its mechanism offer guidance for treating skeletal muscle injuries and supporting animal nutrition research. SFN shows great potential in muscle repair, and future clinical trials could confirm its safety and efficacy, paving the way for new SFN-based treatments and providing new options for patients.

Stomach cancer has become one of the most common types of cancer, with its mortality rate ranking third in the world. Currently, the main treatments for gastric cancer are surgery, radiation therapy, and chemotherapy. Although current treatments can effectively prevent postoperative metastasis and recurrence of gastric cancer, they may also bring various adverse reactions in the gastrointestinal tract and side effects such as bone marrow suppression. Years of research have confirmed that traditional Chinese medicine treatment for gastric and other cancers has distinct characteristics and advantages. Combined treatment can increase the tumor inhibition rate, reduce the side effects of radiation and chemotherapy, improve patients' quality of life, and prolong the survival prognosis.

Gemcitabine (GEM) is used as a first-line therapy for patients diagnosed with any stage of pancreatic cancer (PC); however, patient survival is poor because of GEM resistance. Thus, new approaches to overcome GEM resistance in PC are urgently needed. Here, we aimed to establish an in vivo drug-resistant PC model and identify genes involved in GEM resistance. We focused on one of these factors, CITED4, and elucidated its mechanisms of action in GEM resistance in PC.

Recent research indicates that programmed death 1 (PD-1) and programmed death-ligand 1 (PD-L1) inhibitors show promise in treating triple-negative breast cancer (TNBC), but their efficacy is lower than anticipated, especially when used alone. Therefore, enhancing the anti-tumor immune response strategy for TNBC is crucial. Ubiquitin carboxy-terminal hydrolase L1 (UCH-L1), involved in tumor cell regulation and a potential therapeutic target, has an undefined role in TNBC immunotherapy. In this study, we explored the inverse correlation between UCH-L1 and PD-L1 in TNBC patient tissues. Through in vitro experiments, we found that UCH-L1 negatively regulates PD-L1 by stabilizing the E3 ubiquitin ligase ariadne-1 homolog (ARIH1), which promotes PD-L1 ubiquitination and degradation. Further analysis in Balb/c mice xenograft tumors showed that UCH-L1 correlates with GZMB+/CD8+ T cell infiltration in TNBC, suggesting potential synergistic effects when combining UCH-L1 inhibitors with PD-L1 antibodies. Overall, in TNBC, UCH-L1 stabilizes ARIH1, leading to low PD-L1 expression, which may explain the limited effectiveness of immunotherapy in TNBC patients. Our mouse experiments showed improved therapeutic effects when combining UCH-L1 inhibitors with PD-L1 antibodies. These findings offer a new avenue for immunotherapy in TNBC patients.

(R)-β-homoserine (RBH) is a structural analogue of β-aminobutyric acid (BABA) that can enhance plant resistance to a wide range of pathogens. Here, we investigated the regulatory role of VvTCP7 on the RBH-induced priming response against Botrytis cinerea in grapes. The results showed that RBH primed a defense mechanism in grape berries and enhanced their response to fungal infection. RBH upregulated the expression of a group of genes involved in SA synthesis, thus inducing SA accumulation in grapes. VvTCP7 has high homology to AtCHE in Arabidopsis thaliana and is recognized to be a nucleus-localized protein that promotes SA synthesis. Notably, RBH elevated VvTCP7 expression in harvested grape berries, which was accompanied by enhanced expression of VvNPR1, a master regulator of SAR, as well as the SA-responsive PR genes. Additionally, Y1H, EMSA and DLR assays confirmed that VvTCP7 has the ability to bind directly to the GGNCCC motif within the VvICS promoter to induce VvICS transcription and SA synthesis. Overexpression of VvTCP7 in Arabidopsis led to a marked increase in the transcription of PR genes, enhancing defensive response to B. cinerea. However, the VvTCP7 knockout led to a decrease in PR gene expression and increased susceptibility to the fungus. Collectively, the data suggest that VvTCP7 contributes to RBH-induced SAR priming by activating the SA synthesis and resultant enhances SAR defenses to combat fungal invasion.

We demonstrate that exposure to the AB5 subtilase cytotoxin (SubAB) induces the unfolded protein response (UPR) in human peripheral blood mononuclear cells, concomitant with a proinflammatory response across distinct cell subsets. Notably, SubAB selectively induces type-I interferon (IFN) expression in plasmacytoid dendritic cells, acting synergistically with Toll-like receptor 7 stimulation. The induction of type-I IFN in response to SubAB relies on stimulator of interferon genes (STING) activation, coupled with protein synthesis inhibition mediated by protein kinase R-like endoplasmic reticulum kinase (PERK) and phosphorylation of the eukaryotic translation initiation factor 2 subunit-alpha. By impeding mRNA translation through the integrated stress response, SubAB precipitates the downregulation of the negative innate signaling feedback regulator Tax1-binding protein 1. This downregulation is necessary to unleash TANK-binding kinase 1 signaling associated with STING activation. These findings shed light on how UPR-inducing conditions may regulate the immune system during infection or pathogenesis.

To investigate if FVIII-Fc Fusion protein (FcFVIII) may modulate inflammation and immune stimulation in hemophilic synovium via the Fc-portion of immunoglobulin used for half-life extension we performed gene expression profiling in FVIII-deficient mice. Hemarthrosis was induced by sub-patellar puncture in FVIII-KO mice, + /- periprocedural recombinant human (rh)FVIII,murine (m)FcFVIII, or mIgG2a. Synovium was harvested at baseline and on days (D) 3 and 14, followed by RNA extraction and sequencing, and histological analysis. RNASeq data were processed using standard protocols followed by differential gene expression (DGE) analysis. Functional enrichment analysis generated molecular pathways (KEGG and Reactome). To distinguish between on-target and off-target (related and unrelated to injury/bleed) effects the following groups were compared: i) Baseline vs. injured-saline, ii) injured-saline vs. injured-rhFVIII, iii) injured-saline vs. injured-mFcFVIII. Knee injury in FVIII-KO mice resulted in hemarthrosis, which was prevented by peri-procedural rhFVIII and mFcFVIII treatments. Only a small proportion of genes was affected by FVIII treatment, exhibiting overlap but also distinct differences between both FVIII-preparations. Acutely (D3), mFcFVIII had unique on-target effects related to immune and inflammatory regulation, whereas rhFVIII mostly affected mRNA and protein processing. On day 14, macrophage profiling indicated a transition from M1 to M2, and only mFcFVIII uniquely influenced pathways and genes associated with tissue remodeling and repair. Some mFcFVIII DGE patterns resembled mIgG2a patterns. Synovial vascular remodeling and cartilage health were better with mFcFVIII than rhFVIII. Interestingly, both FVIII-preparations exerted off-target effects on immune system pathways, albeit with temporal differences. These observations provide proof-of-principle that the type of FVIII preparation can influence synovial processes beyond acute hemostasis control, deserving exploration in the setting of joint bleed control in hemophilia.

Acute lung injury (ALI) is a severe inflammatory lung disorder that requires effective therapeutic strategies. Ephedrine (EPH) is the main active component found in medicinal plants of the Ephedra genus and is commonly used to modulate inflammatory responses in various diseases. Hypoxia-inducible factor 1-alpha (HIF1α) is a subunit of hypoxia-inducible factor 1 (HIF1), which plays a critical regulatory role in cellular responses under hypoxic conditions. Moreover, the degradation pathway of HIF1α is regulated by the deubiquitinase Ovarian Tumour Domain-containing Ubiquitin Aldehyde Binding Protein 1 (OTUB1). The aim of this study is to investigate the therapeutic effects of EPH on ALI and its potential therapeutic mechanism. We utilised a lipopolysaccharide (LPS)-induced ALI mouse model and employed various methods for evaluation. Ultimately, our research findings demonstrate that EPH exhibits anti-ALI effects, with the involvement of HIF1α and OTUB1 in the pharmacological actions of EPH. In conclusion, our study results demonstrate that EPH exhibits anti-ALI effects and exerts its protective effects through modulation of the OTUB1 and HIF1α pathways. Our research findings not only lay the foundation for expanding the medicinal applications of EPH but also provide direction for seeking improved treatment strategies for ALI.

Ovarian cancer (OC) remains a leading cause of gynaecological cancer deaths due to late diagnosis and the emergence of resistance to platinum-based chemotherapy, like cisplatin (Cis). Here, we investigated the potential of metformin (Met), a drug commonly used for type 2 diabetes, to overcome Cis resistance in OC. Our findings revealed a synergistic effect of Met with Cis in inhibiting cell viability, proliferation and colony/sphere formation capacity in both cisplatin-sensitive (A2780) and -resistant (A2780/CDDP) ovarian cancer cell lines. This synergistic action triggered apoptosis through DNA damage, S-phase cell cycle arrest and modulation of autophagy. Met also significantly decreased the expression of pluripotency transcription factors (Oct-4, Sox2 and Nanog), indicating its potential to target cancer stem cells (CSCs). Furthermore, the combination therapy downregulated multidrug resistance protein 1 (MDR1) and excision repair cross-complementation group 1 (ERCC1) expression, thereby sensitising resistant cells to Cis-induced cytotoxicity. Additionally, the combination treatment suppressed the Hedgehog (Hh) signalling pathway, which is an important factor in inhibiting CSCs. Our study highlights the potential of the Met signalling pathway to synergise with Cis, overcoming therapeutic resistance in OC by targeting diverse cellular processes, including CSCs, and warrants further investigation in preclinical models.

The combination of gemcitabine and cisplatin serves as a cornerstone in bladder cancer (BC) treatment, yet chemotherapy resistance continues to pose a significant challenge. This study utilizes a novel BC organoid model integrated with drug sensitivity assays to uncover the mechanisms underlying resistance and identify potential therapeutic targets. Our findings reveal that AP1M2 expression is markedly upregulated in gemcitabine- and cisplatin-resistant BC cells and tissues. Elevated AP1M2 levels contribute to enhanced chemotherapy resistance and tumor cell proliferation by facilitating the DNA damage response and increasing RAD54B expression. Mechanistically, AP1M2 interacts with the RNA-binding protein PUM1 to stabilize RAD54B mRNA, thereby supporting DNA repair and survival under chemotherapeutic stress. Notably, inhibition of AP1M2/PUM1-mediated RAD54B expression sensitized BC xenografts to gemcitabine-cisplatin treatment in vivo. These findings unveil a novel mechanism of chemotherapy resistance in BC and highlight the AP1M2/PUM1/RAD54B pathway as a promising therapeutic target to counter resistance and enhance treatment outcomes.

Pseudomonas aeruginosa is an opportunistic pathogen able to form biofilms, contributing to its virulence. With the increasing use of sweeteners in various foods, understanding their influence on bacterial behavior is critical. This study investigated the virulence of P. aeruginosa PAO1 exposed to sweeteners (erythritol, stevia, fructose, coconut sugar, cane sugar, demerara). Sweeteners didn't affect growth rates. Erythritol stimulated biofilm (100 µg/mL, 159.98% formation), while 10 µg/mL of coconut sugar, cane sugar, and demerara promoted lower levels (∼70% formation). Erythritol stimulated exopolysaccharides production but reduced biofilm eDNA. Stevia, fructose, and coconut sugar increased the expression of lasI, lasR, rhlI, rhlR, pqsA, mvfR, and pvdF. HPLC analysis confirmed sucrose as the major sugar in demerara, coconut and cane sugar. Erythritol stimulated biofilm and some virulence genes expression, while other sweeteners' effects varied. Cane sugar was a biofilm inhibitor with a limited gene expression effect. The sweeteners' impact on microorganisms is diverse and should be further investigated.

We investigated the role of Nuclear Receptor Binding SET Domain Protein 2 (NSD2) and microRNAs (miRNAs) in melanoma de-differentiation following Romidepsin and Interferon-α2b (RI) treatment. Melanoma is the most lethal form of skin cancer, and despite advancements in therapy, treatment resistance remains a major challenge. De-differentiation has been widely recognized as a key factor contributing to therapy resistance.

Saline-alkaline stress is a common problem in Akebia trifoliata cultivation. In this study, the enhancing effects of 5-azacytidine (5-AzaC) on the resistance of A. trifoliata to saline-alkaline stress and the underlying mechanisms were investigated. Plant height, stem diameter, biomass, root length, fresh weight of root, and root/shoot ratio of 6-month-old A. trifoliata seedlings were measured after saline-alkaline stress with or without 5-AzaC treatment. Moreover, the contents of photosynthetic pigments, malondialdehyde (MDA), H2O2, sodium, soluble sugar, and proline; activities of superoxide dismutase, peroxidase (POD), and catalase (CAT); and anatomical structures of root, stem, and leaf were assessed. Furthermore, comparative transcriptome sequencing was performed. The results demonstrated that growth and development of A. trifoliata were severely inhibited under saline-alkaline stress, suggesting that the seedlings were exposed to severe oxidative and osmotic stresses. Treatment with exogenous 5-AzaC could significantly relieve the symptoms of saline-alkaline stress in A. trifoliata. Under saline-alkaline stress, 5-AzaC could increase the stem diameter, biomass, root length, fresh weight of root, and root/shoot ratio and minimize damages to the anatomical structure. Moreover, absorption of Na+ was reduced; ionic balance was maintained; POD and CAT activities were significantly improved; proline and soluble sugar contents increased, and H2O2 and MDA contents decreased. Transcriptome analysis revealed that 5-AzaC functioned via regulating KEGG pathways such as plant hormone signal transduction, phenylpropanoid biosynthesis, photosynthesis, amino sugar and nucleotide sugar metabolism, and glutathione metabolism under saline-alkaline stress. Particularly, enhanced expression of genes from the auxin pathway in plant hormone signal transduction; the lignin synthetic pathway in phenylpropanoid biosynthesis; and photosystem II, photosystem I, photosynthetic electron transport, and F-type ATP pathway in photosynthesis may be related to 5-AzaC-induced saline-alkaline resistance. The results provided theoretical references for A. trifoliata cultivation in saline-alkaline soil and application of 5-AzaC to improve saline-alkaline tolerance in plants.

Ellagic acid (EA) exerts anti-carcinogenic activity in various types of cancer. Matrix metalloproteinases (MMPs) are critical mediators in the pathogenesis of renal cell carcinoma (RCC) metastasis. Using in vitro experiments, this study aims to investigate the mechanisms by which EA inhibits RCC migration and invasion. The findings show that EA treatment inhibited RCC cell migration and invasion without reducing cell viability in normal human kidney cells (HK2 cells) and RCC cells (786-O and ACHN). A human proteinase array showed that EA treatment decreased MMP1 mRNA and protein expression levels in 786-O and ACHN cell lines. MMP1 expression is elevated in RCC tissues and correlates with tumor grade, stage, and overall survival in RCC patients. Our molecular docking model indicates a strong interaction between EA and MMP1. The addition of recombinant human MMP1 (Rh-MMP1) to RCC cells increased their migration and invasion; co-treatment with Rh-MMP1 and EA effectively reversed these effects. EA reduced the expression of the transcription factor RUNX2 in both RCC cell lines and knockdown of RUNX2 significantly decreased the migration and invasion abilities of EA-treated 786-O cells. High expression of RUNX2 in RCC patients is associated with higher tumor grade, stage, and poorer survival and correlates positively with MMP1 expression level. These results suggest that EA suppresses RUNX2 targeting of MMP1 expression, thereby conferring anti-invasive properties on RCC cells.

Alyssin is an isothiocyanate found in cruciferous plants. It has been reported to have physiologically active effects. However, no attempts have been made to use alyssin for the treatment of periodontitis. In addition, there are no reports investigating the effects of alyssin on periodontal tissue constituent cells. In the present study, experiments were carried out to determine whether alyssin has an anti-inflammatory effect on human periodontal ligament cells (HPDLCs). We found that alyssin suppressed the interleukin (IL)-1β-induced production of IL-6, IL-8, C-C motif chemokine ligand (CCL)2, CCL20, and C-X-C motif chemokine ligand (CXCL)10 and the expression of intercellular adhesion molecule (ICAM)-1 and cyclooxygenase (COX)-2 in HPDLCs. It was also found that alyssin inhibited the IL-1β-induced activation of nuclear factor (NF)-κB and the phosphorylation of p70S6 kinase (p70S6K) in HPDLCs. Furthermore, we found that the expression of the antioxidant enzymes hemeoxygenase (HO)-1 and NAD(P)H:quinone oxidoreductase (NQO)-1 was enhanced by alyssin treatment of HPDLC. This study suggests that by suppressing the activation of NF-κB and p70S6K, alyssin may attenuate the expression of inflammatory mediators in HPDLCs. In addition, alyssin also increased the expression of antioxidant enzymes.

Pristimerin, which is one of the compounds present in Celastraceae and Hippocrateaceae, has antitumor effects. However, its mechanism of action in esophageal squamous cell carcinoma (ESCC) remains unclear. This study aims to investigate the efficacy and mechanism of pristimerin on ESCC in vitro and in vivo. The inhibitory effect of pristimerin on cell growth was assessed using trypan blue exclusion and colony formation assays. Cell apoptosis was evaluated by flow cytometry. Gene and protein expressions were analyzed through quantitative reverse transcription-polymerase chain reaction (qRT-PCR), Western blotting, and immunohistochemistry. RNA sequencing (RNA-Seq) was employed to identify significantly differentially expressed genes (DEGs). Cell transfection and RNA interference assays were utilized to examine the role of key proteins in pristimerin?s effect. Xenograft models were established to evaluate the antitumor efficiency of pristimerin in vivo. Pristimerin inhibited cell growth and induced apoptosis in ESCC cells. Upregulation of Noxa was crucial for pristimerin-induced apoptosis. Pristimerin activated the Forkhead box O3a (FoxO3a) signaling pathway and triggered FoxO3a recruitment to the Noxa promoter, leading to Noxa transcription. Blocking FoxO3a reversed pristimerin-induced Noxa upregulation and cell apoptosis. Pristimerin treatment suppressed xenograft tumors in nude mice, but these effects were largely negated in Noxa-KO tumors. Furthermore, the chemosensitization effects of pristimerin in vitro and in vivo were mediated by Noxa. This study demonstrates that pristimerin exerts an antitumor effect on ESCC by inducing AKT/FoxO3a-mediated Noxa upregulation. These findings suggest that pristimerin may serve as a potent anticancer agent for ESCC treatment.

Mitomycin C (MC) is an anticancer drug used to treat stomach, anal and lung cancers. The main cytotoxicity of MC is due to its ability to form interstrand crosslinks with DNA (ICLs). The stereochemical configuration at C1″ of MC major ICL is R (α-ICL). In contrast, decarbamoylmitomycin C, a synthetic derivative of MC, generates the major S stereoisomeric ICL (β-ICL). Here, we investigated the effect of the stereochemical configuration of the α/β-ICL on the cellular response by focusing on gene expression changes in MCF-7 and K562 cell lines, one with wild type and the other with mutated TP53, upon treatment with both ICLs. We transfected both cell lines with duplex oligonucleotides containing either the α- or β-ICL at a single site and extracted RNA for transcriptome analysis. Results show that the stereochemical configuration of the α/β-ICL is responsible for distinct gene expression changes in MCF-7 and K562 cells. Our data also show that, in MCF-7 cells, α-ICL treatment triggers a strong increase in CDKN1A expression which is also observed at the protein level, contrary to what happens upon β-ICL treatment. In addition, β-ICL treatment led to a strong downregulation of a greater number of genes than the α-ICL in both cell lines, in particular in K562 cells, which harbor a TP53 mutation. This suggests that the β-ICL toxicity relies on a mechanism which leads to an overall downregulation of gene expression and may explain the greater toxicity of DMC toward TP53 mutant cells.

Glioblastoma (IDH-wildtype) (GBM), the most common high-grade glioma, is a highly invasive and malignant tumor in the brain. Currently, there is no effective treatment for GBM, highlighting the urgent need to find novel therapeutic drugs. Forsythoside B (FB), as a phenylethanoid glycosides compound extracted from Forsythia suspensa, has shown pharmacological functions such as anti-inflammation and anti-bacteria. However, its effects and mechanisms in GBM remain unclear. By performing several in vitro assays, we found that FB suppressed the proliferation of GBM cells in dose- and time-dependent manners. Furthermore, FB arrested the cell cycle at the G0/G1 phase and induced apoptosis in GBM cells. FB also significantly inhibited GBM cells migration. Mechanistically, RNA sequencing results showed that FB treatment remarkably upregulated the expression of PTPRN (a protein-coding gene that plays an important role in the progression of various cancers) in GBM cells. Consistent with this finding, PTPRN expression was downregulated in GBM samples from the Chinese Glioma Genome Atlas (CGGA) and other databases. Knockdown of PTPRN partially restored the inhibitory effects of FB on GBM cells, whereas, overexpression of PTPRN enhanced FB-induced suppression of GBM cell growth and migration. Finally, we found that FB slowed down the growth of tumor in a GBM orthotopic mice model through upregulating PTPRN expression in vivo, with no significant toxicity to other organs. Taken together, these results suggest that FB exerts its anticancer effects on GBM via increasing the expression of PTPRN, which may provide a potential new therapeutic strategy for the treatment of GBM.

Head and neck squamous cell carcinomas (HNSCCs) frequently harbor alterations in the PI3K signalling axis and, particularly, in the PIK3CA gene. The promising rationale of using PI3K inhibitors for the treatment of HNSCC has, however, clashed with the spontaneous development of resistance over time.