Cyclin B2 impairs the p53 signaling in nasopharyngeal carcinoma.

BACKGROUND: Cyclin B2 (CCNB2), a member of the cyclin family, is an oncogene in multiple cancers, including nasopharyngeal carcinoma (NPC). However, the epigenetics mechanism for CCNB2 overexpression in NPC remains unclear. This study dissects the regulatory role of CCNB2 in NPC and the molecular mechanism. METHODS: Differentially methylated genes (DMG) and differentially expressed genes (DEG) were screened out in GSE52068 and GSE13597 databases, respectively, and candidate targets were identified by the Venn diagram. GO annotation and pathway enrichment analyses were performed on selected DMG and DEG, and a PPI network was constructed to pinpoint hub genes. PCR and qMSP were conducted to detect the expression and methylation of CCNB2 in cells. The siRNA targeting CCNB2 was transfected into NPC cells, and the migration, proliferation, cell cycle, epithelial-mesenchymal transition (EMT), tumorigenesis, and metastasis were examined. The upstream factor responsible for CCNB2 overexpression in NPC was explored. The p53 activity in NPC cells was assessed using western blot analysis. RESULTS: CCNB2 showed hypomethylation and overexpression in NPC. CCNB2 silencing inhibited cell migration, proliferation, cell cycle entry, and EMT. JMJD6 was overexpressed in NPC and upregulated CCNB2 through demethylation. JMJD6 reversed the effects of CCNB2 downregulation, resulting in elevated cellular activity in vitro and tumorigenic and metastatic activities in vivo. CCNB2 blocked the p53 pathway, while the p53 pathway inhibitor reversed the effect of CCNB2 silencing to increase the activity of NPC cells. CONCLUSIONS: JMJD6 enhanced CCNB2 transcription by demethylating CCNB2, thereby repressing the p53 pathway and promoting NPC progression.

Bacteria Exploit Autophagy for Proteasome Degradation and Enhanced Virulence in Plants.

Autophagy and the ubiquitin-proteasome system (UPS) are two major protein degradation pathways implicated in the response to microbial infections in eukaryotes. In animals, the contribution of autophagy and the UPS to antibacterial immunity is well documented and several bacteria have evolved measures to target and exploit these systems to the benefit of infection. In plants, the UPS has been established as a hub for immune responses and is targeted by bacteria to enhance virulence. However, the role of autophagy during plant-bacterial interactions is less understood. Here, we have identified both pro- and antibacterial functions of autophagy mechanisms upon infection of Arabidopsis thaliana with virulent Pseudomonas syringae pv tomato DC3000 (Pst). We show that Pst activates autophagy in a type III effector (T3E)-dependent manner and stimulates the autophagic removal of proteasomes (proteaphagy) to support bacterial proliferation. We further identify the T3E Hrp outer protein M1 (HopM1) as a principle mediator of autophagy-inducing activities during infection. In contrast to the probacterial effects of Pst-induced proteaphagy, NEIGHBOR OF BRCA1-dependent selective autophagy counteracts disease progression and limits the formation of HopM1-mediated water-soaked lesions. Together, we demonstrate that distinct autophagy pathways contribute to host immunity and bacterial pathogenesis during Pst infection and provide evidence for an intimate crosstalk between proteasome and autophagy in plant-bacterial interactions.

Transcriptional stimulation of rate-limiting components of the autophagic pathway improves plant fitness.

Autophagy is a major catabolic process whereby autophagosomes deliver cytoplasmic content to the lytic compartment for recycling. Autophagosome formation requires two ubiquitin-like systems conjugating Atg12 with Atg5, and Atg8 with lipid phosphatidylethanolamine (PE), respectively. Genetic suppression of these systems causes autophagy-deficient phenotypes with reduced fitness and longevity. We show that Atg5 and the E1-like enzyme, Atg7, are rate-limiting components of Atg8-PE conjugation in Arabidopsis. Overexpression of ATG5 or ATG7 stimulates Atg8 lipidation, autophagosome formation, and autophagic flux. It also induces transcriptional changes opposite to those observed in atg5 and atg7 mutants, favoring stress resistance and growth. As a result, ATG5- or ATG7-overexpressing plants exhibit increased resistance to necrotrophic pathogens and oxidative stress, delayed aging and enhanced growth, seed set, and seed oil content. This work provides an experimental paradigm and mechanistic insight into genetic stimulation of autophagy in planta and shows its efficiency for improving plant productivity.

Retromer contributes to immunity-associated cell death in Arabidopsis.

Membrane trafficking is required during plant immune responses, but its contribution to the hypersensitive response (HR), a form of programmed cell death (PCD) associated with effector-triggered immunity, is not well understood. HR is induced by nucleotide binding-leucine-rich repeat (NB-LRR) immune receptors and can involve vacuole-mediated processes, including autophagy. We previously isolated lazarus (laz) suppressors of autoimmunity-triggered PCD in the Arabidopsis thaliana mutant accelerated cell death11 (acd11) and demonstrated that the cell death phenotype is due to ectopic activation of the LAZ5 NB-LRR. We report here that laz4 is mutated in one of three VACUOLAR PROTEIN SORTING35 (VPS35) genes. We verify that LAZ4/VPS35B is part of the retromer complex, which functions in endosomal protein sorting and vacuolar trafficking. We show that VPS35B acts in an endosomal trafficking pathway and plays a role in LAZ5-dependent acd11 cell death. Furthermore, we find that VPS35 homologs contribute to certain forms of NB-LRR protein-mediated autoimmunity as well as pathogen-triggered HR. Finally, we demonstrate that retromer deficiency causes defects in late endocytic/lytic compartments and impairs autophagy-associated vacuolar processes. Our findings indicate important roles of retromer-mediated trafficking during the HR; these may include endosomal sorting of immune components and targeting of vacuolar cargo.

Dual role of targeting NAE1 in nasopharyngeal carcinoma: Antitumor effects yet inducing radiotherapy resistance.

Background and objectives: The inhibitor MLN4924 of Neural Precursor Cell-Expressed Developmentally Down-Regulated 8 (NEDD8) Activating Enzyme 1 (NAE1) has been found to suppress the growth of nasopharyngeal carcinoma (NPC). However, its effect on NPC's radiotherapy sensitivity remains unclear. Methods: By integrating single-cell RNA sequencing and bulk RNA sequencing, we predict the impact of NAE1 on the cell cycle, cell death, and its relationship with radiotherapy sensitivity and prognosis in NPC. The effect of inhibiting NAE1 on NPC cell behavior and radiation sensitivity is explored through MLN4924 intervention in vitro and in vivo. We construct a prognosis prediction model based on NAE1 using machine learning methods and validate the efficacy of NAE1 and the model in clinical cohorts. Results: NPC patients with high NAE1 expression have better prognosis and higher expression in the radiotherapy-sensitive group. Inhibiting NAE1 with MLN4924 causes cell cycle arrest in NPC cells, preventing them from entering the G2/M phase, thereby inhibiting proliferation but not affecting migration and metastasis. However, in vitro and in vivo experiments demonstrate that inhibiting NAE1 with MLN4924 leads to increased resistance of NPC to radiation. Conclusions: Targeting NAE1 for NPC treatment may have dual effects, inhibiting NPC proliferation while also increasing radiation resistance.

[Effects of surgical operation on helper-inducer T-lymphocytes in sino-nasal neoplasms patients].

OBJECTIVE: To explore the effects of surgical operation on helper-inducer T-lymphocytes (Th1/Th2) in sino-nasal neoplasms. METHODS: From January 2004 to December 2011, 80 patients with malignant tumor in nasal cavities or sinuses were enrolled as experimental group and another 80 subjects with deflection of nasal septum as control group. The phorbol-12-myristate-13-acetate (PMA)-stimulated peripheral blood mononuclear cells (PBMC) were obtained from both malignant tumor and normal control tissues. And flow cytometry was used to detect the expression percentages of interleukin-4 (IL-4) in Th2 cell and interferon-gamma (IFN-γ) in Th1 cell at pre and post-operation. RESULTS: The preoperative expression percentage of CD3(+) CD8(-) IL-4(+) cell in malignant tumor patients was higher than that in controls (6.6% ± 1.7% vs 2.8% ± 1.7%, P < 0.05) while the expression percentage of CD3(+) CD8(-) IFN-γ(+) cell was lower (18.7% ± 5.7% vs 59.3% ± 1.5%, P < 0.05). The preoperative expression percentage of CD3(+) CD8(-) IL-4(+) cell was higher than that at postoperation (6.6% ± 1.7 %vs 2.8% ± 1.5%, P < 0.05). And the postoperative expression percentage of CD3(+) CD8(-) IFN-γ(+) cell was higher than that at preoperation (54.0% ± 4.0% vs 18.7% ± 5.7%, P < 0.05). CONCLUSION: Surgery may restore the immune balance in patients with malignant tumors in nasal cavities or sinuses.

Spray-Induced Gene Silencing to Study Gene Function in Phytophthora.

RNA interference (RNAi) is a conserved cellular defense mechanism mediated by double-stranded RNA (dsRNA) that can regulate gene expression through targeted destruction of mRNAs (messenger RNAs). Recent studies have shown that spraying dsRNAs or small RNAs (sRNAs) that target essential genes of pathogens on plant surfaces can confer protection against pests and pathogens. Also called spray-induced gene silencing (SIGS), this strategy can be used for disease control and for transient gene silencing to study the function of genes in plant-pathogen interactions. Furthermore, as sRNAs can move locally, systemically, and cross-kingdom during plant-microbe interactions, SIGS allows quick detection and characterization of gene functions in pathogens and plants.

Twigs of dove tree in high-latitude region tend to increase biomass accumulation in vegetative organs but decrease it in reproductive organs.

Adaptive traits are an important dimension for studying the interactions between rare plants and environment. Although the endangered mechanism of rare plants has been reported in many studies, how their twigs adapt to heterogeneous environments associated with latitude is still poorly known. Dove tree (Davidia involucrata Baill.), a monotypic rare species in China, was employed as a model species in our study, and the differences in functional traits, growth relationships and resource allocation among components of annual twig were investigated in three latitudinal regions (32°19' N, 30°08' and 27°55') in the Sichuan, Southwest China. Compared with low- and middle-latitude regions, the twig diameter in high-latitude region decreased by 36% and 26%, and dry mass decreased by 32% and 35%, respectively. Moreover, there existed an allometric growth between flower mass and stem mass or leaf mass in high-latitude region but an isometric growth in low- and middle-latitude regions. At the flower level, an isometric growth between bract area and flower stalk mass was detected among in three latitudinal regions, and the flower stalk mass in the low-latitude region was higher than in the middle- and high-latitude regions for a given bract area and flower mass. At the leaf level, the growth rate of petiole mass was significantly higher than those of leaf area, lamina mass and leaf mass among three latitudinal regions, and the petiole mass in the low-latitude region was higher than in the other two regions for a given leaf mass. Our research demonstrated that the twigs of dove tree in high-latitude region tend to become smaller, and resource input increase in stems and leaves but decrease in flowers, which reflects that dove tree can adapt to the environmental changes across different latitudes by adjusting phenotypic traits growth and biomass allocation of twigs.

A paralog of the MtN3/saliva family recessively confers race-specific resistance to Xanthomonas oryzae in rice.

Approximately one third of the identified 34 rice major disease resistance (R) genes conferring race-specific resistance to different strains of Xanthomonas oryzae pv. oryzae (Xoo), which causes rice bacterial blight disease, are recessive genes. However, only two of the recessive resistance genes have been characterized thus far. Here we report the characterization of another recessive resistance gene, xa25, for Xoo resistance. The xa25, localized in the centromeric region of chromosome 12, mediates race-specific resistance to Xoo strain PXO339 at both seedling and adult stages by inhibiting Xoo growth. It encodes a protein of the MtN3/saliva family, which is prevalent in eukaryotes, including mammals. Transformation of the dominant Xa25 into a resistant rice line carrying the recessive xa25 abolished its resistance to PXO339. The encoding proteins of recessive xa25 and its dominant allele Xa25 have eight amino acid differences. The expression of dominant Xa25 but not recessive xa25 was rapidly induced by PXO339 but not other Xoo strain infections. The nature of xa25-encoding protein and its expression pattern in comparison with its susceptible allele in rice-Xoo interaction indicate that the mechanism of xa25-mediated resistance appears to be different from that conferred by most of the characterized R proteins.

Vacuole Integrity Maintained by DUF300 Proteins Is Required for Brassinosteroid Signaling Regulation.

Brassinosteroid (BR) hormone signaling controls multiple processes during plant growth and development and is initiated at the plasma membrane through the receptor kinase BRASSINOSTEROID INSENSITIVE1 (BRI1) together with co-receptors such as BRI1-ASSOCIATED RECEPTOR KINASE1 (BAK1). BRI1 abundance is regulated by endosomal recycling and vacuolar targeting, but the role of vacuole-related proteins in BR receptor dynamics and BR responses remains elusive. Here, we show that the absence of two DUF300 domain-containing tonoplast proteins, LAZARUS1 (LAZ1) and LAZ1 HOMOLOG1 (LAZ1H1), causes vacuole morphology defects, growth inhibition, and constitutive activation of BR signaling. Intriguingly, tonoplast accumulation of BAK1 was substantially increased and appeared causally linked to enhanced BRI1 trafficking and degradation in laz1 laz1h1 plants. Since unrelated vacuole mutants exhibited normal BR responses, our findings indicate that DUF300 proteins play distinct roles in the regulation of BR signaling by maintaining vacuole integrity required to balance subcellular BAK1 pools and BR receptor distribution.