Background/Aim: Berberine (BBR) is known to be effective at inhibiting cell proliferation and promoting apoptosis in various cancer cells. However, the effects of BBR on triple-negative breast cancer (TNBC) cells remain unclear.The aim of this study was to investigate the cell inhibition effects of BBR on different subtypes of TNBC cells. Methods: Using human TNBC cell lines of different subtypes, namely, MDA-MB-231, MDA-MB-468, MDA-MB-453, and BT-549 as in vitro models, antiproliferative effects of BBR were investigated by 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide assay, trypan blue exclusion assay, and clonogenic assay. Furthermore, cell apoptosis and autophagy were analyzed by flow cytometry, immunofluorescent staining, and LC3 Ⅰ/Ⅱ-targeted Western blotting. Various cell growth-related signaling pathways (AKT/ERK/p38) and the expression of proteins present in various cell cycle kinase complexes were analyzed by Western blotting. Results: BBR concentration-dependently suppressed cell proliferation in MDA-MB-468 (0, 3, 6, and 12 μM) and MDA-MB-231 (0, 6.25, 12.5, and 25 μM). The inhibitory effect was not brought about by inducing cell apoptosis, necrosis, or autophagy. Cell cycle analysis disclosed an increased S+G2/M fraction among the BBR-treated MDA-MB-231 and MDA-MB-453 cells; while with the BBRtreated MDA-MB-468 and BT-549 lines, an increased G0/G1 fraction was found. In MDA-MB-231 and MDA-MB-453 cells, by Western blotting, BBR decreased the expression of Cyclin A and CDK1, On the other hand, in BBR-treated MDA-MB-468 and BT-549 cells, there was a decrease in Cyclin D and CDK4 expression. Conclusion: Our results demonstrate that the antiproliferation effects of BBR occur via different mechanisms in different subtypes of TNBC cells,Abbreviations: BBR, berberine; BL1, basal-like1; BL2, basal-like2; ER, estrogen receptor; HER2, human epidermal growth factor receptor 2; LAR, luminal androgen receptor; M, mesenchymal; PD-1, programmed death-1; PD-L1, programmed death-1 ligand; PR, progestin receptor; ROS, reactive oxygen species; TNBC, triple-negative breast cancer.

Phosphorus, taken up by plants as inorganic phosphate (Pi), is an essential but often growth-43 limiting mineral nutrient for plants. As part of an orchestrated response to improve its 44 acquisition, insufficient Pi supply triggers alterations in root architecture and epidermal cell 45 morphogenesis. Mutants defective in the expression of the OVARIAN TUMOR CONTAINING DEUBIQUITINATING ENZYME5 (OTU5) exhibited a constitutive Pi deficiency 47 root phenotype, comprising the formation of long and dense root hairs and attenuated primary 48 root growth. Quantitative protein profiling of otu5 and wild-type roots using the iTRAQ (isobaric 49Tag for Relative and Absolute Quantification) methodology revealed genotype-and Pi-50 dependent alterations in protein profiles. In otu5 plants, Pi starvation caused a short root hair 51 phenotype and decreased abundance of a suite of Pi-responsive root hair-related proteins. Mutant 52 plants also showed accumulation of proteins involved in chromatin remodeling and altered 53 distribution of reactive oxygen species along the root, which may be causative for the alterations 54 in root hair morphogenesis. The root hair phenotype of otu5 was synergistic to that of arp6, 55 harboring a mutation in the SWR1 chromatin remodeling complex. Genetic analysis of otu5 arp6 56 double mutants suggests independent but functionally related roles of the two proteins in 57 chromatin organization. The root hair phenotype of otu5 is not caused by a general up-regulation 58 of the Pi starvation response, indicating that OTU5 acts downstream of or is interacting with Pi 59 signaling. It is concluded that OTU5 is involved in the interpretation of environmental 60 information, probably by altering chromatin organization and maintaining redox homeostasis. 4 INTRODUCTION 62Phosphorus is an essential structural component of nucleic acids, ATP and cell membranes 63 and plays pivotal roles in signaling and catalytic reactions in metabolism. In plants, phosphorus 64 is taken up as phosphate (Pi), which has limited phyto-availability in most ecosystems and 65 agricultural settings due to complex interactions with soil constituents. Plants, in particular 66 genera that do not engage in mycorrhizal symbiosis, have evolved sophisticated strategies to 67 cope with low Pi availability involving metabolic, physiologic and developmental responses, 68 which aid in improving the acquisition, uptake, and distribution of Pi . 69Due to limited diffusion and mass flow of Pi in the soil solution, increasing the absorptive 70 surface of the root is an efficient strategy to improve Pi acquisition and uptake. In Arabidopsis, 71Pi starvation induces coordinated alterations in root architecture that comprises attenuation of 72 primary root growth and an increase in lateral root formation, a response that has been referred to 73 as topsoil foraging. The restricted longitudinal elongation of epidermal cells increases the 74 number of root hairs per unit root length in permissive positions (over anticlinal walls of cortical 75 cells;...

ORCID IDs: 0000-0002-4868-8284 (D.-F.S.); 0000-0002-7850-6832 (W.S.); 0000-0002-7402-3075 (P.-Y.C.).Phosphate (Pi) starvation induces a suite of adaptive responses aimed at recalibrating cellular Pi homeostasis. Plants harboring a mutation in OVARIAN TUMOR DOMAIN-CONTAINING DEUBIQUITINATING ENZYME5 (OTU5) showed altered DNA methylation of root hair-related genes and altered Pi-responsive root traits. Unlike the wild type, homozygous otu5 mutants did not respond to Pi starvation by increased lateral root formation and increased root hair length but formed very short root hairs when grown on low-Pi media. Under Pi-replete conditions, otu5 plants developed more root hairs than the wild type due to attenuated primary root growth, a phenotype that resembled that of Pi-deficient plants. Growth of plants on low-Pi media altered both H3K4 and H3K27 trimethylation levels at the transcriptional start site of a subset of genes encoding key players in Pi homeostasis, which was correlated with mRNA abundance changes of these genes. Pi starvation had a minor impact on DNA methylation. Differentially methylated regions were enriched in transposable elements, suggesting that DNA methylation associated with low Pi supply is required for maintaining genome integrity. It is concluded that DNA methylation and histone methylation constitute critical, interdependent regulatory components that orchestrate the activity of a subset of Pi-responsive genes.

Manganese (Mn) is pivotal for plant growth and development, but little information is available regarding the strategies that evolved to improve Mn acquisition and cellular homeostasis of Mn. Using an integrated RNA-based transcriptomic and high-throughput shotgun proteomics approach, we generated a comprehensive inventory of transcripts and proteins that showed altered abundance in response to Mn deficiency in roots of the model plant Arabidopsis. A suite of 22,385 transcripts was consistently detected in three RNA-seq runs; LC-MS/MS-based iTRAQ proteomics allowed the unambiguous determination of 11,606 proteins. While high concordance between mRNA and protein expression (R = 0.87) was observed for transcript/protein pairs in which both gene products accumulated differentially upon Mn deficiency, only approximately 10% of the total alterations in the abundance of proteins could be attributed to transcription, indicating a large impact of protein-level regulation. Differentially expressed genes spanned a wide range of biological functions, including the maturation, translation, and transport of mRNAs, as well as primary and secondary metabolic processes. Metabolic analysis by UPLC-qTOF-MS revealed that the steady-state levels of several major glucosinolates were significantly altered upon Mn deficiency in both roots and leaves, possibly as a compensation for increased pathogen susceptibility under conditions of Mn deficiency.

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