Membrane-Bound CD40L Promotes Senescence and Initiates Senescence-Associated Secretory Phenotype via NF-κB Activation in Lung Adenocarcinoma.

BACKGROUND/AIMS: Cellular senescence acts as a barrier against tumorigenesis. The CD40L transgene, expressed in some tumor cells, not only becomes visible to antigen-presenting cells but also actively catalyzes its own termination. Here, we evaluated the effect of a membrane-bound mutant form of human CD40L (CD40L-M) on senescence and the senescence-associated secretory phenotype (SASP) in non-small cell lung cancer (NSCLC). METHODS: CD40 expression levels in the NSCLC cell lines A549/TR, A549/DDP and H460 were examined by flow cytometry. Senescent cells and tissues were identified via SA-ß-gal activity. Cell proliferation was visualized by EdU labeling. qRT-PCR, Western blotting, and immunofluorescence staining were conducted to assess mRNA and protein expression levels of CD40L, γ-H2A.X, p65, p-p65, IκBα, p53, p21 and p16. Cytokines secreted from transfected cells were tested by ELISA and cell migration assay. Capsid tyrosine-modified rAAV5-CD40L-M was packaged and carried out in vivo. RESULTS: Overexpression of CD40L-M promoted senescence, inhibited proliferation, increased DNA damage-associated γ-H2A.X, and initiated the SASP in CD40-positive NSCLC cells. NF-κB signaling was activated by CD40L-M overexpression in these cells. Knockdown of NF-κB partially overcame senescence and failed to induce SASP. Furthermore, increased p53 and p21 protein levels induced by CD40L-M were also reduced following NF-κB suppression. CONCLUSIONS: These data showed that the membrane-bound CD40L mutant may promote cellular senescence and initiate the SASP of NSCLC cells in an NF-κB-dependent manner. Therefore, CD40L-M-induced senescence may be a potential approach to protect against lung adenocarcinoma.

Dynamic DNA Methylation in Plant Growth and Development.

DNA methylation is an epigenetic modification required for transposable element (TE) silencing, genome stability, and genomic imprinting. Although DNA methylation has been intensively studied, the dynamic nature of methylation among different species has just begun to be understood. Here we summarize the recent progress in research on the wide variation of DNA methylation in different plants, organs, tissues, and cells; dynamic changes of methylation are also reported during plant growth and development as well as changes in response to environmental stresses. Overall DNA methylation is quite diverse among species, and it occurs in CG, CHG, and CHH (H = A, C, or T) contexts of genes and TEs in angiosperms. Moderately expressed genes are most likely methylated in gene bodies. Methylation levels decrease significantly just upstream of the transcription start site and around transcription termination sites; its levels in the promoter are inversely correlated with the expression of some genes in plants. Methylation can be altered by different environmental stimuli such as pathogens and abiotic stresses. It is likely that methylation existed in the common eukaryotic ancestor before fungi, plants and animals diverged during evolution. In summary, DNA methylation patterns in angiosperms are complex, dynamic, and an integral part of genome diversity after millions of years of evolution.

[Ozone Deposition and Risk Assessment for a Winter Wheat Field:Partitioning Between Stomatal and Non-stomatal Pathways].

To better understand the ozone deposition and risk assessment over agroecosystems based on the ozone flux indices, an eddy-covariance system was used for measuring the ozone deposition continuously and dynamically in a winter wheat field. We analyzed the variations in ozone concentration, total ozone flux, and stomatal and non-stomatal flux. The relationships between stomatal/non-stomatal ozone deposition velocity and the main meteorological factors were investigated. Finally, the yield losses of winter wheat based on the ozone-dose index (AOT40) and ozone flux index (DFs06) were calculated. Results showed that average daily ozone concentration (cO3) was 32.9 nL·L-1. The daytime (08:00-18:00) and nighttime total ozone flux (FO3) were -7.6 nmol·(m2·s)-1 and -3.1 nmol·(m2·s)-1, respectively, and the mean diurnal FO3 was -5.1 nmol·(m2·s)-1. The mean daily stomatal ozone flux (Fs) and non-stomatal ozone flux (Fns) ranged from 0 to -5.1 nmol·(m2·s)-1 and from -1.43 to -10.31 nmol·(m2·s)-1, respectively. The mean diurnal Fs and Fns were -1.43 nmol·(m2·s)-1 and -3.66 nmol·(m2·s)-1. High solar radiation (SR), high temperature (T), and moderate humidity were used to analyze stomatal ozone deposition; high SR, moderate T, and high humidity were suitable to analyze non-stomatal ozone deposition. The cumulative total ozone flux (DFO3), cumulative stomatal ozone flux (DFs), and cumulative non-stomatal ozone flux (DFns) were 31.58, 9.99, and 21.59 mmol·m-2 during the entire experimental period, and DFs and DFns accounted for 32% and 68% of DFO3. The ranges of yield loss in winter wheat were estimated at 11.58%-20.37% and 20%-23.56% using different assessment models based on the ozone dose index AOT40 and ozone flux index DFs06, respectively.

Effects of Three Protein Phosphatase Inhibitor on A(23187)-induced Apoptosis of HL-60 Cells.

Calcium ionophore A(23187) could increase the intracellular free Ca(2+) concentration and induce apoptosis in some cell lines. In this paper, we reported that A(23187) (1 &mgr;g/ml) could induce apoptosis of HL-60 cells after treating for 4 hours. Pretreatment with the nontoxic concentration of CsA (0.5 3 &mgr;g/ml), an inhibitor of the protein phosphatase 2B (PP2B), could prevent apoptosis induced by A(23187). Neither okadaic acid (OA, inhibitor of PP1, PP2A, PP2C), nor sodium orthovanadate (SoV, inhibitor of tyrosine phosphatase), had such effect. The determination of intracellular Ca(2+) with flow cytometry showed that CsA did not prevent the increase of intracellular Ca(2+) induced by A(23187), which showed that CsA might affect the event in the downstream of calcium increase.