Berberine Induces Caspase-Independent Cell Death in Colon Tumor Cells through Activation of Apoptosis-Inducing Factor

Berberine, an isoquinoline alkaloid derived from plants, is a traditional medicine for treating bacterial diarrhea and intestinal parasite infections. Although berberine has recently been shown to suppress growth of several tumor cell lines, information regarding the effect of berberine on colon tumor growth is limited. Here, we investigated the mechanisms underlying the effects of berberine on regulating the fate of colon tumor cells, specifically the mouse immorto-Min colonic epithelial (IMCE) cells carrying the Apcmin mutation, and of normal colon epithelial cells, namely young adult mouse colonic epithelium (YAMC) cells. Berberine decreased colon tumor colony formation in agar, and induced cell death and LDH release in a time- and concentration-dependent manner in IMCE cells. In contrast, YAMC cells were not sensitive to berberine-induced cell death. Berberine did not stimulate caspase activation, and PARP cleavage and berberine-induced cell death were not affected by a caspase inhibitor in IMCE cells. Rather, berberine stimulated a caspase-independent cell death mediator, apoptosis-inducing factor (AIF) release from mitochondria and nuclear translocation in a ROS production-dependent manner. Amelioration of berberine-stimulated ROS production or suppression of AIF expression blocked berberine-induced cell death and LDH release in IMCE cells. Furthermore, two targets of ROS production in cells, cathepsin B release from lysosomes and PARP activation were induced by berberine. Blockage of either of these pathways decreased berberine-induced AIF activation and cell death in IMCE cells. Thus, berberine-stimulated ROS production leads to cathepsin B release and PARP activation-dependent AIF activation, resulting in caspase-independent cell death in colon tumor cells. Notably, normal colon epithelial cells are less susceptible to berberine-induced cell death, which suggests the specific inhibitory effects of berberine on colon tumor cell growth

Regulatory controls of duplicated gene expression during fiber development in allotetraploid cotton.

Polyploidy complicates transcriptional regulation and increases phenotypic diversity in organisms. The dynamics of genetic regulation of gene expression between coresident subgenomes in polyploids remains to be understood. Here we document the genetic regulation of fiber development in allotetraploid cotton Gossypium hirsutum by sequencing 376 genomes and 2,215 time-series transcriptomes. We characterize 1,258 genes comprising 36 genetic modules that control staged fiber development and uncover genetic components governing their partitioned expression relative to subgenomic duplicated genes (homoeologs). Only about 30% of fiber quality-related homoeologs show phenotypically favorable allele aggregation in cultivars, highlighting the potential for subgenome additivity in fiber improvement. We envision a genome-enabled breeding strategy, with particular attention to 48 favorable alleles related to fiber phenotypes that have been subjected to purifying selection during domestication. Our work delineates the dynamics of gene regulation during fiber development and highlights the potential of subgenomic coordination underpinning phenotypes in polyploid plants. [Abstract copyright: © 2023. The Author(s).

Breast-cancer-secreted miR-122 reprograms glucose metabolism in premetastatic niche to promote metastasis

Reprogrammed glucose metabolism as a result of increased glycolysis and glucose uptake is a hallmark of cancer. Here we show that cancer cells can suppress glucose uptake by non-tumour cells in the pre-metastatic niche, by secreting vesicles that carry high levels of the miR-122 microRNA. High miR-122 levels in the circulation have been associated with metastasis in breast cancer patients and we show that cancer-cell-secreted miR-122 facilitates metastasis by increasing nutrient availability in the pre-metastatic niche. Mechanistically cancer-cell-derived miR-122 suppresses glucose uptake by niche cells in vitro and in vivo by downregulating the glycolytic enzyme pyruvate kinase (PKM). In vivo inhibition of miR-122 restores glucose uptake in distant organs, including brain and lungs, and decreases the incidence of metastasis. These results demonstrate that by modifying glucose utilization by recipient pre-metastatic niche cells, cancer-derived extracellular miR-122 is able to reprogram systemic energy metabolism to facilitate disease progression

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

High-Efficiency Second-Harmonic Generation Using Quasi-Bound State in LiNbO₃ Metasurface

We numerically demonstrated a high-efficiency second-harmonic generation (SHG) using quasi-bound state in the continuum (quasi–BIC) in thin film LiNbO3 (TFLN) metasurface. The TFLN possessed exceptionally high second-order nonlinear coefficients, contributing to the enhanced SHG performance. An eccentric cylinder unit cell was presented to achieve high Q–factor resonances associated with the asymmetric parameter introduced. Simulations showed that the high efficiency of the second-harmonic conversion was obtained by using the high Q–factor of the asymmetric dielectric cylinder metasurface, and it achieved a high SHG efficiency of 6.5% at pump intensities as low as 1 MW/cm2 at a normal incident. Furthermore, the simulation results indicated that breaking the symmetry through oblique incidence was more effective in achieving a higher Q–factor compared to altering the structural parameters. Specifically, under 1° oblique incidences, the conversion efficiency could reach 1.2% at an incident power of 1 kW/cm². We have proposed a method to achieve a high conversion efficiency of second-harmonic generation in low-refractive-index materials. Our work not only offers theoretical support but also provides valuable insights for the advancement of efficient nonlinear frequency doubling technology, optical communication, and sensing applications

Otolith Microchemistry Reveals Life History and Habitat Use of Coilia nasus from the Dayang River of China

The estuarine tapertail anchovy Coilia nasus is distributed throughout the Dayang River. However, the life history and habitat use of this fish remain unknown. Here, the microchemistry patterns of Sr and Ca in 23 otoliths collected from the lower reaches of the Dayang River were analyzed using an X-ray electron probe microanalyzer. The anchovies were divided into two patterns: (1) with low Sr/Ca ratios (&lt;3.0) and a single bluish Sr concentration map, indicating that it only experienced a freshwater habitat during its whole life, and (2) with Sr/Ca ratios fluctuating between low and high (&gt;3.0) phases and Sr concentration maps showing various colors, including blue, green, yellow, and red, from the core to the edge of otoliths, whose larvae hatched in freshwater and spent their first winter in brackish or sea waters. The juveniles then stayed in estuarine water areas for further growth and feeding until sexual maturity, when the mature adults returned to the spawning grounds in the river. The co-existence of freshwater residents and anadromous C. nasus in the Dayang River has been studied for the first time, and its possible spawning ground was discovered. These findings provide essential information to effectively protect this species and guide its rational, sustainable utilization

Revealing Population Connectivity of the Estuarine Tapertail Anchovy Coilia nasus in the Changjiang River Estuary and Its Adjacent Waters Using Otolith Microchemistry

The estuarine tapertail anchovy, Coilia nasus, is a migratory fish with high economic value in China. We collected fish from the Changjiang River (the Yangtze River) estuary, the Qiantang River estuary, and the southern Yellow Sea, and studied their relationships using otolith elemental and stable isotopic microchemistry signatures to assess the population connectivity of C. nasus. Results show that, in addition to Ca, other elements were present in the otolith core. The &delta;18O, Na/Ca, Fe/Ca, and Cu/Ca values of the Qiantang population were significantly higher than those of the others, whereas its &delta;13C and Ba/Ca values were found to be significantly lower. Otolith multi-element composition and stable isotope ratios differed significantly between the Qiantang and Changjiang estuary groups (p &lt; 0.05); however, no difference was observed between the latter and the Yellow Sea group. Cluster analysis, linear discriminant analysis, and a self-organizing map strongly suggest possible connectivity between the fish populations of the Changjiang estuary and Yellow Sea, while the population of the Qiantang River estuary appears to be independent. Notably, results suggest a much closer connectivity between the fish populations of the Changjiang River and the Yellow Sea

Berberine stimulates AIF release from the mitochondria to the cytoplasm and nuclear translocation in IMCE, which is required for berberine-induced cell death.

<p>Cells were treated with berberine at 50 µM for the indicated times, as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036418#pone-0036418-g002" target="_blank">Figure 2</a>. Cytoplasmic, nuclear, and mitochondrial fractions were prepared for Western blot analysis using anti-AIF, anti-GAPDH (cytosolic marker), anti-COX IV (mitochondrial marker) and anti-Ki-67 (nuclear marker) antibodies (A). Immunostaining of cells with 50 µM berberine treatment for 18 h was performed to localize AIF using an anti-AIF antibody and a FITC-conjugated secondary antibody (green) and nuclei using DAPI staining (blue). Arrows indicate nuclei with AIF translocation (B). IMCE cells transfected with AIF siRNA or non-targeting siRNA for 24 h were treated with berberine at 50 µM for 18 h to detect number change (D) and LDH release (E) as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036418#pone-0036418-g002" target="_blank">Figure 2</a>. AIF expression levels were detected by Western blot analysis using an anti-AIF antibody (C). Actin blot was used as a protein loading control. *<i>p</i><0.01 compared to the control group.</p

Berberine activates PARP in IMCE, which mediates berberine-mediated AIF activation and cell death.

<p>Cells were treated with berberine at 50 µM for the indicated times (A) or at the indicated concentrations for 18 h (B). PARP activity was measured using the HT 167 Universal Colorimetric PARP Assay Kit. PARP activation is expressed as a percentage of control in each experiment. Cells were treated with berberine at 50 µM for 18 h in the presence or absence of DPG (25 µM), a PARP activation inhibitor. Cell number change (C) and LDH release (D) were detected as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036418#pone-0036418-g002" target="_blank">Figure 2</a>. The cytosolic fractions were prepared for Western blot analysis of AIF, as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036418#pone-0036418-g003" target="_blank">Figure 3</a> (E). *<i>p</i><0.01 compared to the control group, #<i>p<</i>0.01 compared to the berberine treated group.</p