ZFHX3 acts as a tumor suppressor in prostate cancer by targeting FTO-mediated m6A demethylation.

Zinc-finger homeobox 3 (ZFHX3, also known as ATBF1) suppresses prostatic tumorigenesis. ZFHX3 is frequently found to have numerous deletions in human prostate cancer (PCa). However, the underlying molecular function of ZFHX3 during prostatic tumorigenesis is not well understood. N6-methyladenosine (m6A) modification in RNA plays a critical role in the development of cancers; however, the relationship between ZFHX3 and m6A modification is largely unknown in PCa. In this study, we found that ZFHX3 knockdown decreased total m6A levels through enhancing the transcriptional activity of FTO in PCa cells. Importantly, FTO inhibition suppressed cell proliferation and rescued the promoting function of ZFHX3 knockdown on cell proliferation. In vivo, we verified that FTO was upregulated and ZFHX3 was decreased in PCa patients and that a high level of ZFHX3 is indispensable for low FTO expression and is correlated with better patient survival. Through transcriptome sequencing and MeRIP sequencing, we revealed that E2F2 and CDKN2C were the direct targets of FTO-mediated m6A modification and ZFXH3 was required for the regulation of FTO on E2F2 and CDKN2C expression. Unexpectedly, we uncovered that ZFHX3 expression was in return regulated by FTO in an m6A-dependent way. These findings establish a novel crosstalk mechanism between ZFHX3 and FTO in prostatic tumorigenesis.

Characterization of Pseudomonas aeruginosa bacteriophages and control hemorrhagic pneumonia on a mice model.

Pseudomonas aeruginosa is one of the most common pathogens causing hemorrhagic pneumonia in Chinese forest musk deer. Multidrug-resistant P. aeruginosa is frequently isolated from the lungs of affected musk deer in Shaanxi Province, China. With the increasing bacterial drug resistance, commonly used antibiotics have shown limited efficacy against drug-resistant P. aeruginosa. Therefore, phages have garnered attention as a promising alternative to antibiotics among researchers. In this study, phages vB_PaeP_YL1 and vB_PaeP_YL2 (respectively referred to as YL1 and YL2) were isolated from mixed sewage samples from a farm. YL1 and YL2 exhibit an icosahedral head and a non-contractile short tail, belonging to the Podoviridae family. Identification results demonstrate good tolerance to low temperatures and pH levels, with minimal variation in potency within 30 min of UV irradiation. The MOI for both YL1 and YL2 was 0.1, and their one-step growth curve latent periods were 10 min and 20 min, respectively. Moreover, both single phage and phage cocktail effectively inhibited the growth of the host bacteria in vitro, with the phage cocktail showing superior inhibitory effects compared to the single phage. YL1 and YL2 possess double-stranded DNA genomes, with YL1 having a genome size of 72,187 bp and a total G + C content of 55.02%, while YL2 has a genome size of 72,060 bp and a total G + C content of 54.98%. YL1 and YL2 are predicted to have 93 and 92 open reading frames (ORFs), respectively, and no ORFs related to drug resistance or lysogeny were found in both phages. Genome annotation and phylogenetic analysis revealed that YL1 is closely related to vB_PaeP_FBPa1 (ON857943), while YL2 is closely related to vB_PaeP_FBPa1 (ON857943) and Phage26 (NC041907). In a mouse model of hemorrhagic pneumonia, phage cocktail treatment showed better control of the disease and significantly reduced lung bacterial load compared to single phage treatment. Therefore, YL1 and YL2 have the potential for the prevention and treatment of multidrug-resistant P. aeruginosa infections.

Fluorescent probes for ferroptosis bioimaging: advances, challenges, and prospects.

Ferroptosis is a form of regulatory cell death distinct from caspase-dependent apoptosis and plays an important role in life entities. Since ferroptosis involves a variety of complex regulatory factors, the levels of certain biological species and microenvironments would change during this process. Thus, the investigation of the level fluctuation of key target analytes during ferroptosis is of great significance for disease treatment and drug design. Toward this aim, multiple organic fluorescent probes with simple preparation and non-destructive detection have been developed, and research over the past decade has uncovered a vast array of homeostasis and other physiological characteristics of ferroptosis. However, this significant and cutting-edge topic has not yet been reviewed. In this work, we aim to highlight the latest breakthrough results of fluorescent probes for monitoring various bio-related molecules and microenvironments during ferroptosis at the cellular, tissue and in vivo levels. Accordingly, this tutorial review has been organized according to the target molecules identified by the probes including ionic species, reactive sulfur species, reactive oxygen species, biomacromolecules, microenvironment, and others. In addition to providing new insights into the findings of each fluorescent probe in ferroptosis studies, we also discuss the defects and limitations of the probes developed, and highlight the potential challenges and further prospects in this domain. We anticipate that this review will convey profound implications for designing powerful fluorescent probes to decrypt changes in key molecules and microenvironments during ferroptosis.

Exogenous Arginine Treatment Maintains the Appearance and Nutraceutical Properties of Hard- and Soft-Seed Pomegranates in Cold Storage.

Arginine is a natural preservative; however, its effects on the storage of different cultivars of pomegranates have not been investigated extensively. Therefore, the fruit quality of soft-seed Tunisia and hard-seed Yudazi pomegranates was investigated after treatment with arginine at four concentrations during cold storage for 80 days. Pomegranates treated with 1.0 mM arginine exhibited a relatively lower loss of vitamin C, soluble solid, total phenol, and anthocyanin contents in arils, together with a better fruit appearance. Combined with principal component analysis (PCA), the storage life of fruits treated with 1.0 mM arginine showed a higher correlation with antioxidant enzyme activity (e.g., superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT)) during the first 40 days of cold storage, whereas after 40 days of cold storage, storage life was more dependent on the integrity of the cell membrane affected by malondialdehyde (MDA) content, electrolyte leakage (EL), and hydrogen peroxide (H2O2) accumulation. Arginine treatment contributed significantly to the appearance and inner quality of the hard-seed pomegranate cv. Yudazi fruit during cold storage compared to those of soft-seed Tunisia. Taken together, arginine application combined with cold storage enhanced the nutraceutical properties and marketability of pomegranate fruits.

Genome-Wide Identification and Comprehensive Analysis of the AP2/ERF Gene Family in Pomegranate Fruit Development and Postharvest Preservation.

Pomegranate (Punica granatum L.) is a kind of fruit with significant economic, ecological and health values. AP2/ERF transcription factors belong to a large group of factors mainly found in plants and play key roles in plant growth and development. However, AP2/ERF genes in pomegranate and their implication in development and postharvest preservation have been little described. In this study, 116 PgAP2/ERF genes in pomegranate were identified and renamed based on their chromosomal distributions. Phylogenetic relationship with genes from other species, structures, duplications, annotations, cis-elements in promoter sequences, and protein-protein interaction networks among PgAP2/ERF proteins were comprehensively explored. Expression analysis revealed several PgAP2/ERFs associated with the phenotypes of pomegranate seed hardness, including PgAP2/ERF5, PgAP2/ERF36, PgAP2/ERF58, and PgAP2/ERF86. Subsequent analysis indicated that many differentially expressed PgAP2/ERF genes are potentially important regulators of pomegranate fruit development. Furthermore, expression of more than one-half of PgAP2/ERFs was repressed in 'Tunisian soft seed' pomegranate fruit under low-temperature cold storage. The results showed that 1-MCP implicated in promoting postharvest preservation of 'Tunisian soft seed' pomegranate upregulated the PgAP2/ERF4, PgAP2/ERF15, PgAP2/ERF26, PgAP2/ERF30, PgAP2/ERF35 and PgAP2/ERF45 genes compared to those under low-temperature cold storage. This indicates that these genes are important candidate genes involved in pomegranate postharvest preservation. In summary, the findings of the present study provide an important basis for characterizing the PgAP2/ERF family genes and provide information on the candidate genes involved in pomegranate fruit development and postharvest preservation.

Zfhx3 is essential for progesterone/progesterone receptor signaling to drive ductal side-branching and alveologenesis in mouse mammary glands.

Progesterone (Pg)/progesterone receptor (PR) signaling drives mammary gland side-branching and alveologenesis, but the mechanisms through which Pg/PR signaling functions remain to be clarified. Using in vitro and in vivo models and histological and molecular analyses, we determined the role of Zfhx3 transcription factor in mammary gland development driven by Pg/PR signaling. Postnatal deletion of Zfhx3 in mouse mammary epithelial cells attenuated side-branching morphogenesis and alveologenesis. These effects were undetectable in the absence of Pg/PR signaling. During the estrus cycle, Zfhx3 expression corresponded to that of Pg, being at the highest level at the diestrus stage; Zfhx3 deletion inhibited mammary gland branching more potently at diestrus than estrus stage. Loss of Zfhx3 not only attenuated the expansion of stem/progenitor cells driven by Pg/PR signaling, but also impaired the function of Pg/PR signaling in the transcriptional activation of multiple genes. In addition, Pg/PR signaling significantly expanded PR- and Zfhx3-positive epithelial cells, and induced the physical association of ZFHX3 with PR. These findings establish Zfhx3 as an integral transcription factor of Pg/PR signaling in driving side-branching and alveologenesis during mammary gland development.

ZFHX3 is indispensable for ERß to inhibit cell proliferation via MYC downregulation in prostate cancer cells.

Both estrogen receptor 2 (ESR2, also known as estrogen receptor beta (ERß)) and the zinc-finger homeobox 3 (ZFHX3, also known as ATBF1 for AT motif-binding factor 1) modulate prostate development and suppress prostatic tumorigenesis in mice. ZFHX3 is integral to proper functions of ESR1 (i.e., estrogen receptor alpha (ERα)), which belongs to the same family of proteins as ESR2, but is hardly expressed in prostate epithelial cells. It is not clear how ZFHX3 suppresses prostatic tumorigenesis. In this study, we investigated whether ZFHX3 and ERß functionally interact with each other in the suppression of prostatic tumorigenesis. In two androgen receptor (AR)-positive prostate cancer cell lines, C4-2B and LNCaP, we first validated ERß's tumor suppressor activity indicated by the inhibition of cell proliferation and repression of MYC expression. We found that loss of ZFHX3 increased cell proliferation and MYC expression, and downregulation of MYC was necessary for ZFHX3 to inhibit cell proliferation in the same cell lines. Importantly, loss of ZFHX3 prevented ERß from suppressing cell proliferation and repressing MYC transcription. Biochemically, ERß and ZFHX3 physically interacted with each other and they both occupied the same region of the common MYC promoter, even though ZFHX3 also bound to another region of the MYC promoter. Higher levels of ZFHX3 and ERß in human prostate cancer tissue samples correlated with better patient survival. These findings establish MYC repression as a mechanism for ZFHX3's tumor suppressor activity and ZFHX3 as an indispensable factor for ERß's tumor suppressor activity in prostate cancer cells. Our data also suggest that intact ZFHX3 function is required for using ERß-selective agonists to effectively treat prostate cancer.

LEM4 confers tamoxifen resistance to breast cancer cells by activating cyclin D-CDK4/6-Rb and ERα pathway.

The elucidation of molecular events that confer tamoxifen resistance to estrogen receptor α (ER) positive breast cancer is of major scientific and therapeutic importance. Here, we report that LEM4 overexpression renders ER+ breast cancer cells resistant to tamoxifen by activating the cyclin D-CDK4/6 axis and the ERα signaling. We show that LEM4 overexpression accelerates tumor growth. Interaction with LEM4 stabilizes CDK4 and Rb, promotes Rb phosphorylation and the G1/S phase transition. LEM4 depletion or combined tamoxifen and PD0332991 treatment significantly reverses tamoxifen resistance. Furthermore, LEM4 interacts with and stabilizes both Aurora-A and ERα, promotes Aurora-A mediated phosphorylation of ERα-Ser167, leading to increase in ERα DNA-binding and transactivation activity. Elevated levels of LEM4 correlates with poorer relapse-free survival in patients with ER+ breast cancer undergoing endocrine therapy. Thus, LEM4 represents a prognostic marker and an attractive target for breast cancer therapeutics. Functional antagonism of LEM4 could overcome tamoxifen resistance.

Demineralized Bone Scaffolds with Tunable Matrix Stiffness for Efficient Bone Integration.

As a biophysical cue, matrix stiffness can decide the stem cell fate. However, most methods to construct three-dimensional (3D) scaffolds may change the 3D microstructure while altering their mechanical properties. In this study, demineralized bone matrix scaffolds with different compressive modulus (66.06 ± 27.83 MPa (high), 26.90 ± 13.16 MPa (medium), and 0.67 ± 0.14 MPa (low)) were constructed by controlling the decalcification duration (1 h, 12 h, and 5 days), respectively. The pore size and porosity have no significant difference between the scaffolds before and after decalcification. Cell experiments indicated that the low scaffolds could promote the osteogenic differentiation of bone marrow mesenchymal stem cells (MSCs) in vitro. Rat subcutaneous implantation experiments further demonstrated that the low scaffolds could efficiently improve the cell infiltration, deposition of collagen fibers, and positive osteocalcin and osteopontin expression of endogenous cells as well as angiogenesis. Finally, rabbit femoral condylar defect experiments proved that the low scaffolds could significantly promote the bone repair and integration and stromal cell derived factor-1α/CXC chemokine receptor signal pathway was essential for the stiffness-mediated bone repair. These investigations provided a novel method for fabricating 3D bone grafts with different stiffness, which is also of great significance for studying the effect of stiffness on the biological behavior of MSCs in three dimensions.

[Structure and biological functions of mammalian LEM-domain proteins].

During the process of open mitosis in higher eukaryotic cells, the nuclear envelope (NE) is disassembled and reassembled with highly organized and periodical dynamic morphological changes. Recent studies demonstrated that LEM-domain protein family mediates interactions among inner nuclear membrane, nuclear lamina protein and chromatin by interacting with barrier-to-autointegration-factor (BAF). The structure and function of the ternary complex formed by LEM-domain protein, nuclear lamina protein and BAF are dependent on each other. Moreover, the network formation based on this structure and function is critical for the development of basic biological processes of nuclear, and it plays important roles in chromatin separation in late metaphase and anaphase, NE reassembly after mitosis, morphological maintenance of nuclear and NE in interphase, regulation of DNA replication and DNA damage repair, regulation of gene expression and signaling pathway, and infection of retrovirus. Mutations in genes encoding LEM family proteins have important impacts on development and progression of laminopathic diseases and tumorigenesis. This review provides a detailed summary of structural and functional studies of the LEM family proteins.