PLOS ONE: [sortOrder=DATE_NEWEST_FIRST, sort=Date, newest first, q=subject:"Cell biology"]PLOShttps://journals.plos.org/plosone/webmaster@plos.orgaccelerating the publication of peer-reviewed sciencehttps://journals.plos.org/plosone/search/feed/atom?sortOrder=DATE_NEWEST_FIRST&unformattedQuery=subject:%22Cell+biology%22&sort=Date,+newest+firstAll PLOS articles are Open Access.https://journals.plos.org/plosone/resource/img/favicon.icohttps://journals.plos.org/plosone/resource/img/favicon.ico2024-03-29T04:47:00ZChronic viral infection impairs immune memory to a different pathogenCheng YangZhicui LiuYing YangLuis J. CockaYongguo LiWeihong ZengHao Shen10.1371/journal.ppat.10121132024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by Cheng Yang, Zhicui Liu, Ying Yang, Luis J. Cocka, Yongguo Li, Weihong Zeng, Hao Shen</p>
Chronic viral infections cause T cell dysfunction in both animal models and human clinical settings, thereby affecting the ability of the host immune system to clear viral pathogens and develop proper virus-specific immune memory. However, the impact of chronic viral infections on the host’s immune memory to other pathogens has not been well described. In this study, we immunized mice with recombinant <i>Listeria monocytogenes</i> expressing OVA (Lm-OVA) to generate immunity to Lm and allow analysis of OVA-specific memory T (Tm) cells. We then infected these mice with lymphocytic choriomeningitis virus (LCMV) strain Cl-13 which establishes a chronic infection. We found that chronically infected mice were unable to protect against <i>Listeria</i> re-challenge. OVA-specific Tm cells showed a progressive loss in total numbers and in their ability to produce effector cytokines in the context of chronic LCMV infection. Unlike virus-specific T cells, OVA-specific Tm cells from chronically infected mice did not up-regulate the expression of inhibitory receptors, a hallmark feature of exhaustion in virus-specific T cells. Finally, OVA-specific Tm cells failed to mount a robust recall response after bacteria re-challenge both in the chronically infected and adoptively transferred naïve hosts. These results show that previously established bacteria-specific Tm cells become functionally impaired in the setting of an unrelated bystander chronic viral infection, which may contribute to poor immunity against other pathogens in the host with chronic viral infection.Behind the scenes: Centromere-driven genomic innovations in fungal pathogensAswathy NarayananMd. Hashim RezaKaustuv Sanyal10.1371/journal.ppat.10120802024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by Aswathy Narayanan, Md. Hashim Reza, Kaustuv Sanyal</p>The <i>Wolbachia</i> WalE1 effector alters <i>Drosophila</i> endocytosisMaryAnn MartinSergio López-MadrigalIrene L. G. Newton10.1371/journal.ppat.10112452024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by MaryAnn Martin, Sergio López-Madrigal, Irene L. G. Newton</p>
The most common intracellular bacterial infection is <i>Wolbachia pipientis</i>, a microbe that manipulates host reproduction and is used in control of insect vectors. Phenotypes induced by <i>Wolbachia</i> have been studied for decades and range from sperm-egg incompatibility to male killing. How <i>Wolbachia</i> alters host biology is less well understood. Previously, we characterized the first <i>Wolbachia</i> effector–WalE1, which encodes an alpha-synuclein domain at the N terminus. Purified WalE1 sediments with and bundles actin and when heterologously expressed in flies, increases <i>Wolbachia</i> titer in the developing oocyte. In this work, we first identify the native expression of WalE1 by <i>Wolbachia</i> infecting both fly cells and whole animals. WalE1 appears as aggregates in the host cell cytosol. We next show that WalE1 co-immunoprecipitates with the host protein Past1, although might not directly interact with it, and that WalE1 manipulates host endocytosis. Yeast expressing WalE1 show deficiency in uptake of FM4-64 dye, and flies harboring mutations in <i>Past1</i> or overexpressing WalE1 are sensitive to AgNO<sub>3</sub>, a hallmark of endocytosis defects. We also show that flies expressing WalE1 suffer from endocytosis defects in larval nephrocytes. Finally, we also show that <i>Past1</i> null flies harbor more <i>Wolbachia</i> overall and in late egg chambers. Our results identify interactions between <i>Wolbachia</i> and a host protein involved in endocytosis and point to yet another important host cell process impinged upon by <i>Wolbachia’s</i> WalE1 effector.Characterization of the complete mitochondrial genome of <i>Desmaulus extinctorium</i> (Littorinimorpha, Calyptraeoidea, Calyptraeidae) and molecular phylogeny of LittorinimorphaYanwen MaBiqi ZhengJiji LiWei MengKaida XuYingying Ye10.1371/journal.pone.03013892024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by Yanwen Ma, Biqi Zheng, Jiji Li, Wei Meng, Kaida Xu, Yingying Ye</p>
For the purpose of determining the placement of Calyptraeidae within the Littorinimorpha, we hereby furnish a thorough analysis of the mitochondrial genome (mitogenome) sequence of <i>Desmaulus extinctorium</i>. This mitogenome spans 16,605 base pairs and encompasses the entire set of 37 genes, including 13 PCGs, 22 tRNAs and two rRNAs, with an evident AT bias. Notably, <i>tRNA</i><sup>Ser1</sup> and <i>tRNA</i><sup>Ser2</sup> lack dihydrouracil (DHU) arms, resulting in an inability to form a secondary structure. Similarly, <i>tRNA</i><sup>Ala</sup> lacks a TΨC arm, rendering it incapable of forming a secondary structure. In contrast, the remaining tRNAs demonstrate a characteristic secondary structure reminiscent of a cloverleaf. A comparison with ancestral gastropods reveals distinct differences in three gene clusters (or genes), encompassing 15 tRNAs and eight PCGs. Notably, inversions and translocations represent the major types of rearrangements observed in <i>D</i>. <i>extinctorium</i>. Phylogenetic analysis demonstrates robust support for a monophyletic grouping of all Littorinimorpha species, with <i>D</i>. <i>extinctorium</i> representing a distinct Calyptraeoidea clade. In summary, this investigation provides the first complete mitochondrial dataset for a species of the Calyptraeidae, thus providing novel insights into the phylogenetic relationships within the Littorinimorpha.Quantitative imaging and semiotic phenotyping of mitochondrial network morphology in live human cellsSophie CharrasseVictor RacineCharlotte Saint-OmerTitouan PoquillonLoïc LionnardMarine LedruChristophe GonindardSandrine DelaunoisKarima KissaRichard E. FryeManuela PastoreChristelle ReynesMathilde FrechetHanane ChajraAbdel Aouacheria10.1371/journal.pone.03013722024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by Sophie Charrasse, Victor Racine, Charlotte Saint-Omer, Titouan Poquillon, Loïc Lionnard, Marine Ledru, Christophe Gonindard, Sandrine Delaunois, Karima Kissa, Richard E. Frye, Manuela Pastore, Christelle Reynes, Mathilde Frechet, Hanane Chajra, Abdel Aouacheria</p>
The importance of mitochondria in tissue homeostasis, stress responses and human diseases, combined to their ability to transition between various structural and functional states, makes them excellent organelles for monitoring cell health. There is therefore a need for technologies to accurately analyze and quantify changes in mitochondrial organization in a variety of cells and cellular contexts. Here we present an innovative computerized method that enables accurate, multiscale, fast and cost-effective analysis of mitochondrial shape and network architecture from confocal fluorescence images by providing more than thirty features. In order to facilitate interpretation of the quantitative results, we introduced two innovations: the use of Kiviat-graphs (herein named MitoSpider plots) to present highly multidimensional data and visualization of the various mito-cellular configurations in the form of morphospace diagrams (called MitoSigils). We tested our fully automated image analysis tool on rich datasets gathered from live normal human skin cells cultured under basal conditions or exposed to specific stress including UVB irradiation and pesticide exposure. We demonstrated the ability of our proprietary software (named MitoTouch) to sensitively discriminate between control and stressed dermal fibroblasts, and between normal fibroblasts and other cell types (including cancer tissue-derived fibroblasts and primary keratinocytes), showing that our automated analysis captures subtle differences in morphology. Based on this novel algorithm, we report the identification of a protective natural ingredient that mitigates the deleterious impact of hydrogen peroxide (H2O2) on mitochondrial organization. Hence we conceived a novel wet-plus-dry pipeline combining cell cultures, quantitative imaging and semiotic analysis for exhaustive analysis of mitochondrial morphology in living adherent cells. Our tool has potential for broader applications in other research areas such as cell biology and medicine, high-throughput drug screening as well as predictive and environmental toxicology.Short-term chemotherapy-related complications and undernutrition in children diagnosed with cancer at Korle Bu Teaching Hospital, Accra, Ghana: A prospective cohort studyNihad SalifuCatherine I. SegbefiaYakubu AlhassanLorna A. RennerEdem M. A. Tette10.1371/journal.pone.03012082024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by Nihad Salifu, Catherine I. Segbefia, Yakubu Alhassan, Lorna A. Renner, Edem M. A. Tette</p>
Undernutrition in children with cancer is associated with complications during cancer therapy. The study objective was to determine the association between specific anthropometric parameters and short-term chemotherapy-related complications and mortality. This was a hospital-based, prospective cohort study of children, age ≤12 years, with a new cancer diagnosis at the Paediatric Oncology Unit, Korle Bu Teaching Hospital, Ghana. Socio-demographic information, cancer characteristics and anthropometric measurements were obtained at enrolment. Participants were followed up for twelve weeks from commencement of chemotherapy and selected treatment-related complications such as anaemia and thrombocytopenia requiring transfusions, prolonged neutropenia resulting in treatment delays, febrile neutropenia, mucositis and death were recorded. A total of 133 participants were recruited with a median age of 4.5 years. Eighty-one (60.9%) were diagnosed with solid tumours, 31 (23.3%) had leukaemias and 21 (15.8%) had lymphomas. Of the anthropometric parameters assessed, only arm anthropometry using upper arm muscle area (UAMA) and mid-upper arm circumference (MUAC) were associated with complications. Participants with wasting were more likely to develop anaemia and mucositis. However, the incidence of prolonged neutropenia was significantly higher among participants with average UAMA (p = 0.043) and low average UAMA (p = 0.049) compared to those with low UAMA. Risk of neutropenia was also significantly less among those with wasting by MUAC compared to those well-nourished (p = 0.045). Twenty-three participants (17.3%) died with a greater proportion (11/44; 25%) occurring in those who were wasted using MUAC. These findings underscore the need for nutritional surveillance at diagnosis and during chemotherapy, particularly where co-morbid disease is prevalent.Piperine alleviates nonalcoholic steatohepatitis by inhibiting NF-κB-mediated hepatocyte pyroptosisSuye RanLingyu SongHong YangJiangnan YuYunhuan ZhenQi Liu10.1371/journal.pone.03011332024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by Suye Ran, Lingyu Song, Hong Yang, Jiangnan Yu, Yunhuan Zhen, Qi Liu</p>
Purpose <p>Nonalcoholic steatohepatitis (NASH) is the progressive form of nonalcoholic fatty liver disease (NAFLD), which has a high risk of cirrhosis, liver failure, and hepatocellular carcinoma. Piperine (Pip) is an extract of plants with powerful anti-inflammatory effects, however, the function of Pip in NASH remains elusive. Here, we aim to explore the role of Pip in NASH and to find the possible mechanisms.</p> Methods <p>Methionine and choline-deficient (MCD) diets were used to induce steatohepatitis, methionine- and choline-sufficient (MCS) diets were used as the control. After Pip treatment, H&E staining, Oil Red O staining, hepatic triglyceride (TG) content and F4/80 expression were performed to analysis liver steatosis and inflammation; Masson’s staining, COL1A1 and α-SMA were detected liver fibrosis. Lipopolysaccharide (LPS) -treated AML12 cells were used to as the cell model to induce pyroptosis. Then, pyroptosis-related proteins, IL-1β and LDH release were detected <i>in vivo</i> and <i>in vitro</i>. Finally, NF-κB inhibitor, BAY11-7082, was used to further demonstrate the mechanism of Pip in NASH.</p> Results <p>The study found that Pip alleviated liver steatosis, inflammation, hepatocyte injury, and fibrosis in mice fed with MCD diets. Moreover, the pyroptosis markers (NLRP3, ASC, caspase-1 p20, and GSDMD), IL-1β and LDH release were decreased by Pip treatment. NF-κB activation was suppressed by Pip treatment and pyroptosis-related proteins were down regulated by BAY11-7082.</p> Conclusion <p>Pip ameliorates NASH progression, and the therapeutical effect was associated with inhibition of hepatocyte pyroptosis induced by NF-κB.</p>Stochastic modeling of a gene regulatory network driving B cell development in germinal centersAlexey KoshkinUlysse HerbachMaría Rodríguez MartínezOlivier GandrillonFabien Crauste10.1371/journal.pone.03010222024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by Alexey Koshkin, Ulysse Herbach, María Rodríguez Martínez, Olivier Gandrillon, Fabien Crauste</p>
Germinal centers (GCs) are the key histological structures of the adaptive immune system, responsible for the development and selection of B cells producing high-affinity antibodies against antigens. Due to their level of complexity, unexpected malfunctioning may lead to a range of pathologies, including various malignant formations. One promising way to improve the understanding of malignant transformation is to study the underlying gene regulatory networks (GRNs) associated with cell development and differentiation. Evaluation and inference of the GRN structure from gene expression data is a challenging task in systems biology: recent achievements in single-cell (SC) transcriptomics allow the generation of SC gene expression data, which can be used to sharpen the knowledge on GRN structure. In order to understand whether a particular network of three key gene regulators (BCL6, IRF4, BLIMP1), influenced by two external stimuli signals (surface receptors BCR and CD40), is able to describe GC B cell differentiation, we used a stochastic model to fit SC transcriptomic data from a human lymphoid organ dataset. The model is defined mathematically as a piecewise-deterministic Markov process. We showed that after parameter tuning, the model qualitatively recapitulates mRNA distributions corresponding to GC and plasmablast stages of B cell differentiation. Thus, the model can assist in validating the GRN structure and, in the future, could lead to better understanding of the different types of dysfunction of the regulatory mechanisms.Transcriptomics analysis of the bovine endometrium during the perioestrus periodMohammed A. AlfattahCarolina N. CorreiaJohn A. BrownePaul A. McGettiganKatarzyna PlutaStephen D. CarringtonDavid E. MacHughJane A. Irwin10.1371/journal.pone.03010052024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by Mohammed A. Alfattah, Carolina N. Correia, John A. Browne, Paul A. McGettigan, Katarzyna Pluta, Stephen D. Carrington, David E. MacHugh, Jane A. Irwin</p>
During the oestrous cycle, the bovine endometrium undergoes morphological and functional changes, which are regulated by alterations in the levels of oestrogen and progesterone and consequent changes in gene expression. To clarify these changes before and after oestrus, RNA-seq was used to profile the transcriptome of oestrus-synchronized beef heifers. Endometrial samples were collected from 29 animals, which were slaughtered in six groups beginning 12 h after the withdrawal of intravaginal progesterone releasing devices until seven days post-oestrus onset (luteal phase). The groups represented proestrus, early oestrus, metoestrus and early dioestrus (luteal phase). Changes in gene expression were estimated relative to gene expression at oestrus. Ingenuity Pathway Analysis (IPA) was used to identify canonical pathways and functional processes of biological importance. A total of 5,845 differentially expressed genes (DEGs) were identified. The lowest number of DEGs was observed at the 12 h post-oestrus time point, whereas the greatest number was observed at Day 7 post-oestrus onset (luteal phase). A total of 2,748 DEGs at this time point did not overlap with any other time points. Prior to oestrus, <i>Neurological disease</i> and <i>Organismal injury and abnormalities</i> appeared among the top IPA diseases and functions categories, with upregulation of genes involved in neurogenesis. Lipid metabolism was upregulated before oestrus and downregulated at 48h post-oestrus, at which point an upregulation of immune-related pathways was observed. In contrast, in the luteal phase the <i>Lipid metabolism</i> and <i>Small molecule biochemistry pathways</i> were upregulated.Characterization of a hemolytic and antibiotic-resistant <i>Pseudomonas aeruginosa</i> strain S3 pathogenic to fish isolated from Mahananda River in IndiaDipanwita GhoshPreeti MangarAbhinandan ChoudhuryAnoop KumarAniruddha SahaProtip BasuDipanwita Saha10.1371/journal.pone.03001342024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by Dipanwita Ghosh, Preeti Mangar, Abhinandan Choudhury, Anoop Kumar, Aniruddha Saha, Protip Basu, Dipanwita Saha</p>
Virulent strain <i>Pseudomonas aeruginosa</i> isolated from Mahananda River exhibited the highest hemolytic activity and virulence factors and was pathogenic to fish as clinical signs of hemorrhagic spots, loss of scales, and fin erosions were found. S3 was cytotoxic to the human liver cell line (WRL-68) in the trypan blue dye exclusion assay. Genotype characterization using whole genome analysis showed that S3 was similar to <i>P</i>. <i>aeruginosa</i> PAO1. The draft genome sequence had an estimated length of 62,69,783 bp, a GC content of 66.3%, and contained 5916 coding sequences. Eight genes across the genome were predicted to be related to hemolysin action. Antibiotic resistance genes such as class C and class D beta-lactamases, <i>fosA</i>, <i>APH</i>, and <i>catB</i> were detected, along with the strong presence of multiple efflux system genes. This study shows that river water is contaminated by pathogenic <i>P</i>. <i>aeruginosa</i> harboring an array of virulence and antibiotic resistance genes which warrants periodic monitoring to prevent disease outbreaks.Changes in the intestinal microbiota of individuals with non-alcoholic fatty liver disease based on sequencing: An updated systematic review and meta-analysisWenpin CaiTing QiuWeitao HuTaiyong Fang10.1371/journal.pone.02999462024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by Wenpin Cai, Ting Qiu, Weitao Hu, Taiyong Fang</p>
Background <p>Alterations in the composition and abundance of the intestinal microbiota occur in non-alcoholic fatty liver disease (NAFLD). However, the results are inconsistent because of differences in the study design, subject area, and sequencing methodology. In this study, we compared the diversity and abundance of the intestinal microbiota of patients with NAFLD and healthy individuals through a systematic review and meta-analysis.</p> Methods <p>Three databases (PubMed, EMBASE, and Cochrane Library) were searched from their inception to March 20, 2023. A meta-analysis was performed using Stata software to analyze variations in the richness and abundance of the intestinal microbiota in patients with NAFLD. The Newcastle-Ottawa Quality Assessment Scale (NOS) was used for quality assessment.</p> Results <p>A total of 28 articles were included. Shannon diversity was reduced in patients with NAFLD (SMD = -0.24 (95% CI -0.43–0.05, I<sup>2</sup> = 71.7%). The relative abundance of <i>Ruminococcus</i>, <i>Faecalibacterium</i>, and <i>Coprococcus</i> all decreased, with total SMDs of -0.96 (95% CI -1.29 to -0.63, I<sup>2</sup> = 4.8%), -1.13 (95% CI -2.07 to -0.19, I<sup>2</sup> = 80.5%), and -1.66 (95% CI -3.04 to -0.28, I<sup>2</sup> = 91.5%). <i>Escherichia</i> was increased in individuals with NAFLD (SMD = 1.78, 95% CI 0.12 to 3.45, I<sup>2</sup> = 94.4%).</p> Conclusion <p>Increasing the species diversity and altering the abundance of specific gut microbiota, including <i>Coprococcus</i>, <i>Faecalibacterium</i>, <i>Ruminococcus</i>, and <i>Escherichia</i>, may be beneficial for improving NAFLD.</p>The effect of cooperator recognition on competition among clones in spatially structured microbial communitiesAdrienna BinghamAparajita SurLeah B. ShawHelen A. Murphy10.1371/journal.pone.02995462024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by Adrienna Bingham, Aparajita Sur, Leah B. Shaw, Helen A. Murphy</p>
In spatially structured microbial communities, clonal growth of stationary cells passively generates clusters of related individuals. This can lead to stable cooperation without the need for recognition mechanisms. However, recent research suggests that some biofilm-forming microbes may have mechanisms of kin recognition. To explore this unexpected observation, we studied the effects of different types of cooperation in a microbial colony using spatially explicit, agent-based simulations of two interacting strains. We found scenarios that favor a form of kin recognition in spatially structured microbial communities. In the presence of a “cheater” strain, a strain with greenbeard cooperation was able to increase in frequency more than a strain with obligate cooperation. This effect was most noticeable in high density colonies and when the cooperators were not as abundant as the cheaters. We also studied whether a polychromatic greenbeard, in which cells only cooperate with their own type, could provide a numerical benefit beyond a simple, binary greenbeard. We found the greatest benefit to a polychromatic greenbeard when cooperation is highly effective. These results suggest that in some ecological scenarios, recognition mechanisms may be beneficial even in spatially structured communities.Describe the morphology and mitochondrial genome of <i>Mecidea indica</i> Dallas, 1851 (Hemiptera, Pentatomidae), with its phylogenetic positionChao ChenDongmei BaiZhenhua ZhangXiaofei DingShuzhen YangQing ZhaoHufang Zhang10.1371/journal.pone.02992982024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by Chao Chen, Dongmei Bai, Zhenhua Zhang, Xiaofei Ding, Shuzhen Yang, Qing Zhao, Hufang Zhang</p>
We here describe the external morphology and complete mitochondrial genome characteristics of <i>Mecidea indica</i> Dallas, 1851, and clarify the evolutionary rate and divergence time. The <i>M</i>. <i>indica</i> mitochondrial genome length is 15,670 bp, and it exhibits a typical high A+T-skew (76.31%). The sequence shows strong synteny with the original gene arrangement of <i>Drosophila yakuba</i> Burla, 1954 without rearrangement. The <i>M</i>. <i>indica</i> mitochondrial genome characteristics were analyzed, and phylogenetic trees of Pentatomidae were reconstructed using Bayesian methods based on different datasets of the mitochondrial genome datasets. Phylogenetic analysis shows that <i>M</i>. <i>indica</i> belongs to Pentaotominae and form a sister-group with <i>Anaxilaus musgravei</i> Gross, 1976, and Asopinae is highly supported as monophyletic. Molecular clock analysis estimates a divergence time of Pentatomidae of 122.75 Mya (95% HPD: 98.76–145.43 Mya), within the Mesozoic Cretaceous; the divergence time of <i>M</i>. <i>indica</i> and <i>A</i>. <i>musgravii</i> was no later than 50.50 Mya (95% HPD: 37.20–64.80 Mya). In addition, the divergence time of Asopinae was 62.32 Mya (95% HPD: 47.08–78.23 Mya), which was in the Paleogene of the Cenozoic era. This study is of great significance for reconstructing the phylogeny of Pentatomidae and providing insights into its evolutionary history.Transcriptome-wide analysis of the differences between MCF7 cells cultured in DMEM or αMEMYang JiaoHongbo ZhaoLin LuXiangyu ZhaoYanchun WangBingrong Zheng10.1371/journal.pone.02982622024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by Yang Jiao, Hongbo Zhao, Lin Lu, Xiangyu Zhao, Yanchun Wang, Bingrong Zheng</p>
MCF7 cells have been used as an experimental model for breast cancer for decades. Typically, a culture medium is designed to supply cells with the nutrients essential for their continuous proliferation. Each medium has a specific nutritional composition. Therefore, cells cultured in different media may exhibit differences in their metabolism. However, only a few studies have investigated the effects of media on cells. In this study, we compared the effects of Dulbecco’s modified Eagle medium (DMEM) and minimum essential medium alpha modification (αMEM) on MCF7 cells. The two media differentially affected the morphology, cell cycle, and proliferation of MCF7 cells, but had no effect on cell death. Replacement of DMEM with αMEM led to a decrease in ATP production and an increase in reactive oxygen species production, but did not affect the cell viability. RNA-sequencing and bioinformatic analyses revealed 721 significantly upregulated and 1247 downregulated genes in cells cultured in αMEM for 48 h compared with that in cells cultured in DMEM. The enriched gene ontology terms were related to mitosis and cell proliferation. Kyoto encyclopedia of genes and genomes analysis revealed cell cycle and DNA replication as the top two significant pathways. MCF7 cells were hypoxic when cultured in αMEM. These results show that the culture medium considerably affects cultured cells. Thus, the stability of the culture system in a study is very important to obtain reliable results.17β-Estradiol promotes metastasis in triple-negative breast cancer through the Calpain/YAP/β-catenin signaling axisXuemei NiuJianan WangJinguang LiuQinglong YuMingwei Ci10.1371/journal.pone.02981842024-03-28T14:00:00Z2024-03-28T14:00:00Z<p>by Xuemei Niu, Jianan Wang, Jinguang Liu, Qinglong Yu, Mingwei Ci</p>
β-catenin is an important regulator of malignant progression. 17β-Estradiol (E2), an important sex hormone in women, promotes the growth and metastasis of triple-negative breast cancer (TNBC). However, whether β-catenin is involved in E2-induced metastasis of TNBC remains unknown. In this study, we show that E2 induces the proliferation, migration, invasion, and metastasis of TNBC cells. E2 induces β-catenin protein expression and nuclear translocation, thereby regulating the expression of target genes such as Cyclin D1 and MMP-9. The inhibition of β-catenin reversed the E2-induced cell malignant behaviors. Additionally, E2 activated Calpain by increasing intracellular Ca<sup>2+</sup> levels and reducing calpastatin levels. When Calpain was inhibited, E2 did not induce the proliferation, migration, invasion, or metastasis of TNBC cells. In addition, E2 promoted translocation of YAP into the nucleus by inhibiting its phosphorylation. Calpain inhibition reversed the E2-induced YAP dephosphorylation. Inhibition of YAP transcriptional activity reversed the effects of E2 on the proliferation, migration, invasion, and β-catenin of TNBC cells. In conclusion, we demonstrated that E2 induced metastasis-related behaviors in TNBC cells and this effect was mediated through the Calpain/YAP/β-catenin signaling pathway.