Histochemistry and Cell Biology最新文献

Fanconi anemia (FA) is primarily an autosomal recessive genetic disorder that leads to bone marrow failure, increased risk of developing cancer, and a plethora of developmental abnormalities. Patients are prone to recurrent infections and increased risk of hemorrhage, as well as delayed wound healing with poor results. FA is caused by a genetic mutation in the proteins needed for FA pathway activation; FA group D2 protein (FANCD2) is an indispensable part of this pathway and plays essential roles in some aspects of cellular life, especially in the cellular responses to DNA damage. Here, we found that depletion of FANCD2 induced reduction of proliferation and migration of NIH3T3 cells. Moreover, FANCD2 knockout decreased production of extracellular matrix (ECM) protein collagen III and cytoskeleton protein alpha-smooth muscle actin (α-SMA). In this process, FANCD2 knockout decreased the expression of DNA methyltransferase 1 (DNMT1), and DNMT1 inhibitor 5-aza-2'-deoxycytidine (5-AZA-CdR) also induced the decline of proliferation and migration ability of NIH3T3 cells, and reduced the expression of collagen III and α-SMA. These findings suggest that FANCD2 affects wound healing through DNMT1. These findings may provide novel therapeutic ideas for clinical treatment of patients with FA with poor wound healing.

The adrenal glands are essential endocrine organs whose cortex and medulla maintain systemic homeostasis and mediate stress responses via steroid hormone and catecholamine secretion. Despite anatomical and functional similarities between human and mouse adrenal glands, notable species-specific differences exist. Here, we leveraged spatial transcriptomics (10× Genomics Visium) to comprehensively map gene expression in adult human and mouse adrenal glands, aiming to identify canonical marker genes conserved across species. The analysis was based on a 31-year-old female human sample (GEO dataset) and four 10-week-old male CD-1 mice. Human adrenal sections were processed using optimal cutting temperature (OCT) embedding, whereas mouse adrenal sections were processed as formalin-fixed paraffin-embedded (FFPE) samples, highlighting differences in sample preparation. Using unsupervised clustering of spatial gene expression data, we delineated distinct adrenal cortex and medulla zones in both species, confirming known zonation patterns. Our cross-species analysis revealed highly conserved spatial expression of key known marker genes characteristic of the adrenal cortex (e.g., CYP11B2 for ZG, CYP11B1 for ZF) and medullary chromaffin cells (e.g., TH), as well as a core set of additional marker genes previously less characterized in adrenal biology. By integrating transcriptional profiles, we generated a catalogue of conserved canonical marker genes that define adrenal zonation and function in both humans and mice. These results highlight the fundamental molecular conservation of adrenal gland organization and support the translational value of mouse models in adrenal research. Our findings provide new insights into the evolutionary preservation of adrenal function and a valuable resource for studies on adrenal physiology and disease.

The incidence of colorectal cancer (CRC) and the associated mortality in CRC patients have been rising in recent years. Quiescin sulfhydryl oxidase 1 (QSOX1), a secreted disulfide catalyst essential for extracellular matrix (ECM) assembly, is upregulated in several tumors (e.g. pancreatic, breast, and lung cancer), often correlating with aggressive tumor phenotypes and worse prognosis. In contrast, colorectal and hepatocellular carcinoma specimens show significant downregulation of QSOX1 compared to normal or adjacent tissue. Recognizing cancer as a heterocellular tissue where stromal cell types are crucial to tumor behavior, we evaluated by immunohistochemistry stromal and epithelial QSOX1 expression in 140 CRC cases (mean age: 64 years, 56% female, 83% without neoadjuvant therapy) and 10 normal colon samples. We found that stromal QSOX1 expression is significantly reduced in CRC compared to normal colon tissue (p < 0.0001). The stromal, rather than the epithelial, compartment determines total QSOX1 levels in tumor samples. Stromal QSOX1 demonstrates a negative relation with tumor size (ß = -0.04, p < 0.05), but not with other histopathological characteristics. This finding suggests that stromal QSOX1 loss may play a key role in early tumor cell proliferation rather than in clinical progression. Importantly, stromal QSOX1 expression shows excellent discriminatory power between tumor and non-tumor tissues, with an AUC of 0.98 in ROC analysis. Altogether, QSOX1 downregulation is a defining feature of tumor-associated stroma in CRC, likely affecting ECM integrity and modulating epithelial-stromal crosstalk. Thus, further characterization of stromal molecular signatures may identify novel biomarkers and therapeutic target strategies in CRC.

In the human placenta, trophoblast cells give rise to two unique cell layers that envelop the surface of chorionic villi: the outer syncytiotrophoblast (STB) layer and the inner cytotrophoblast (CTB) cell layer. The structural changes in CTB cells during gestation are still not fully understood. This study examined the ultrastructural integrity of the CTB layer within chorionic villi of the human term placenta using Fast Red immunohistochemistry in conjunction with proteinase K/Triton X-100 tissue-clearing and serial block-face scanning electron microscopy (SBF-SEM). Visualization of the CTB layer in whole-mount peripheral villous trees was facilitated by Fast Red immunohistochemistry of SPINT1, a marker specific to CTB cells, under both bright-field and fluorescence imaging modes in light microscopy. CTB cells displayed a thin, flattened morphology and extended multiple cellular projections, resulting in a spider-like transformation that envelops grape-like terminal villi. In contrast, in more proximal villi (e.g., stem villi), CTB cells showed a thick, cuboidal, or polygonal appearance, covering the villous subsurface. SBF-SEM imaging demonstrated the structure of a thin, mesh-like CTB layer, where the basal domain of the villous surface STB infiltrated through the small gaps of the CTB layer and contacted fetal capillaries via the basal lamina in terminal villi. Our data suggest that terminal villi undergo structural changes to facilitate fetomaternal exchange.

The expression of different sarco/endoplasmic reticulum calcium ATPase (SERCA) isoforms is controversial in various cancers and is not clear in the experimental cancer model. The present study attempts to evaluate the expression dynamics of SERCA isoforms in the azoxymethane/dextran sulphate sodium salt (AOM/DSS) model of colorectal carcinogenesis in mice. Inflammation-associated colorectal cancer was induced in the mice by administration of a single dose of AOM and three alternative cycles of DSS in drinking water. Body weights were recorded weekly. Mice were killed at weeks 0, 8, 12 and 16. At those times, the number of tumours was recorded, and colon tissues were processed for histopathological, immunohistochemical and gene expression analysis. The number of tumours and the formation of aberrant crypt foci were found to be significantly higher in the AOM/DSS group compared to the control. Histopathology of the colon revealed a higher percentage of dysplasia, adenoma and adenocarcinoma formation in the AOM/DSS group, further supported by high intensity of immunohistochemical staining for PCNA in the same. Gene expression analysis indicated higher expression of cyclin D1, β-catenin and low expression of E-cadherin, suggesting carcinogenic transformation of the colon. Immunohistochemical and gene expression analysis of SERCA isoforms indicated higher expression of SERCA1 and SERCA2 and low expression of SERCA3 in colon tissues of the AOM/DSS-exposed animals. The present study confirmed a similar expression pattern of SERCA isoforms in the AOM/DSS model of carcinogenesis as reported in clinical samples. Further, this study highlights the fact that altered SERCA patterns could be a contributing factor in the development of colorectal carcinogenesis.

Telocytes (TCs) are interstitial cells characterized by long, thin cytoplasmic extensions called telopodes. Although their presence has been established in various organs, data on their existence and organization within the human ovary remain limited. This study aimed to identify and describe telocytes in the ovarian stroma using immunohistochemical, immunofluorescence, and ultrastructural methods. Ovarian tissues from women aged 24-65 years who underwent total hysterectomy and bilateral salpingo-oophorectomy for nonmalignant uterine diseases were analyzed. TCs were identified by CD34 co-expression with c-KIT, vimentin, platelet-derived growth factor receptor (PDGFR)-β, and alpha-smooth muscle actin (α-SMA) and confirmed by transmission electron microscopy with immunogold labeling. The results demonstrated that telocytes form two to three discontinuous layers around the adventitia of large and medium-sized blood vessels and establish close contacts with stromal components, including smooth muscle cells. This spatial organization suggests their involvement in intercellular communication and stromal coordination within the ovarian microenvironment. These findings provide the first ultrastructural and immunohistochemical evidence of telocytes in the human ovarian stroma and highlight the need for further studies to clarify their physiological and pathological roles in ovarian function, including potential morphological and molecular differences among females of different age groups.

Neoadjuvant chemoradiotherapy (nCRT) followed by surgery is the standard treatment for locally advanced rectal cancer. However, the response to nCRT is variable and not always associated with improved survival. Colorectal cancer (CRC) is characterized by a complex tumor microenvironment (TME), a key component of which is cancer-associated fibroblasts (CAF). While their functions and interactions with tumor cells are under active investigation, the role of distinct CAF subpopulations and their plasticity remains largely undefined. A comprehensive characterization of CAFs during cancer progression could therefore contribute to the development of novel anticancer diagnostic and therapeutic strategies. In this study, we used immunohistochemistry to demonstrate that nCRT induces a significant reorganization of the TME in rectal adenocarcinoma. This reorganization was characterized by a redistribution of myofibroblasts (alpha-smooth muscle actin, αSMA⁺) and tumor-associated fibroblasts (fibroblast activation protein, FAP⁺; and fibroblast-specific protein 1, FSP1⁺), leading to pronounced fibrosis in both central and peripheral tumor regions. A decrease in E-cadherin expression, coupled with increased vimentin and transforming growth factor beta (TGFβ) levels in a subset of patients, indicated activation of epithelial-mesenchymal transition and the emergence of vasculogenic mimicry as an alternative mechanism of tumor vascularization. Ultrastructural analysis revealed structural changes in the cytoplasm of fibroblasts suggestive of active membrane remodeling and fibroblast-myofibroblast transition, particularly at the tumor periphery. Collectively, these findings suggest that nCRT may be accompanied by a reorganization of the tumor stroma, leading to fibroblast activation, epithelial-mesenchymal transition, and vasculogenic mimicry, all of which could potentially contribute to tumor progression.

Oxidative stress plays a crucial role in ovarian physiology and pathology, yet its dynamic changes across the estrous cycle remain unclear. This study aimed to investigate oxidative stress in rat ovarian tissue during the stages of the estrous cycle through histological and biochemical analyses. Vaginal smears identified estrous cycle stages, and ovarian tissues were examined histologically under a light microscope. Follicle counts, corpus luteum (CL) classification, oxidative stress marker (8-hydroxy-2'-deoxyguanosine/8-OHdG) staining, and apoptotic activity (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labeling/TUNEL) staining were analyzed. Serum malondialdehyde (MDA) and superoxide dismutase (SOD) activity was measured. Results showed strong 8-OHdG positivity in follicular epithelial cells surrounding the oocyte, granulosa cells of antral follicles, and CL cells. The detection of 8-OHdG positivity in CL cells, in both the nuclei and cytoplasm, suggests that oxidative stress affects both nuclear and mitochondrial DNA. Serum MDA levels were highest during estrus, while SOD activity was highest during metestrus. The increase in oxidative stress was associated with ovulation during estrus, while variations in SOD activity reflected changing defense mechanisms throughout the cycle. 8-OHdG positivity was high in follicular and luteal cells during CL regression, indicating ROS impact on follicular development and luteal function. Apoptotic cells were present mainly in antral follicles and luteal cells. These findings highlight the critical role of oxidative stress in ovarian function, with potential implications for fertility regulation and reproductive medicine. With rising oxidative stress levels, alternative cell death mechanisms likely contribute alongside apoptosis during CL regression.