Cytojournal最新文献

Objective: Breast fine-needle aspiration cytology (FNAC) is a rapid, cost-effective, and minimally invasive diagnostic procedure. The diagnosis of a proliferative breast disease with atypia (PBDA) is established based on the presence of areas with disordered cellular arrangement and mildly discerned cytological features. We have aimed to explore the cytohistological correlation of PBDA on FNAC.

Material and methods: A review of the hospital database was undertaken to retrieve cases of breast FNAC diagnosed as PBDA between January 2011 and September 2020.

Results: A total of 3125 breast FNAC specimens were examined, and 107 (3.4%) of them received the diagnosis of PBDA. A total of 68 PBDA cases were included in this cytohistological evaluation. The risk of malignancy was 44%. Except for one case, all of the invasive or microinvasive carcinomas were grade 1 or 2 malignancies according to the Nottingham grading system of breast cancers. The result of a repeat FNAC of the case with a poorly differentiated invasive breast cancer was reported as a high-grade malignancy. A statistically significant correlation was observed between older age and malignant outcome (P < 0.001).

Conclusion: This is one of the largest datasets of cases with PBDA. Based on the advanced age of the patient, and relevant clinical and radiological information, cytopathological diagnosis of PBDA may prompt the clinician to take further action.

Objective: Hepatocellular carcinoma (HCC) represents a primary liver tumor characterized by rapid disease progression and unfavorable clinical outcomes. Most patients with HCC are identified in advanced stage, where targeted therapies are considered an effective treatment method for advanced disease. The tyrosine kinase inhibitor (TKI) donafenib has shown efficacy in managing HCC. However, drug resistance often occurs after treatment with donafenib, which limits its widespread clinical application. Thus, this study aims to identify small-molecule TKIs that can enhance the sensitivity of HCC to donafenib.

Material and methods: The HCC cells HepG2 and SNU449 were treated with five drugs, namely, dimethyl sulfoxide, AZ-628, SU-5402, TG-101209, and SPP-86, combined with donafenib to determine half-maximal inhibitory concentration values. RNA sequencing data obtained from The Cancer Genome Atlas (TCGA) were analyzed using Differential Expression analysis for Sequence data 2 (DESeq2)/limma and Gene Set Enrichment Analysis (GSEA). The effects of AZ-628 on proliferation, viability, apoptosis, and migration were assessed. The expression level of early growth response gene 1 (EGR1) was measured through Western blotting/quantitative polymerase chain reaction and silenced by cell transfection. Donafenib-resistant HepG2 cells negative control shRNA (shNC)/shRNA targeting EGR1 (shEGR1) were treated with AZ-628 combined with donafenib. Ferrous ion (Fe²⁺) and reactive oxygen species levels were measured after Erastin/RSL3 induction. The synergy between AZ-628 and donafenib was analyzed using Combenefit2. In vivo, tumor growth, and Ki67 expression were evaluated in nude mice treated with DMSO, AZ-628, donafenib, or their combination.

Results: This study showed that AZ-628 reduced donafenib resistance in HCC by targeting the tyrosine kinase (TK) pathway. Cell counting kit-8 and colony formation assay validated that AZ-628 significantly improved the sensitivity of HCC cells to donafenib (P < 0.0001). Rescue experiments showed that AZ-628 regulated HCC cell proliferation and drug resistance through EGR1 (P < 0.001). In addition, AZ-628 was found to affect donafenib resistance in HCC by regulating epithelial-mesenchymal transition, apoptosis, and ferroptosis (P < 0.0001). In vivo experiments demonstrated a combined anti-tumor efficacy of AZ-628 and donafenib in HCC models (P < 0.0001).

Conclusion: The findings of this study reveal a new combination therapy targeting the TK pathway for the treatment of HCC and provide a theoretical foundation for addressing donafenib resistance.

Objective: The function of tumor protein p53-inducible nuclear protein 2 (TP53INP2) in numerous cancers has been elucidated, but its role across the development of spinal cord glioma (SCG) remains largely unexplored. This study aims to explore the anti-cancer effect of TP53INP2/nuclear factor-kappa B (NF-kB)/nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway in SCG.

Material and methods: Quantitative real-time polymerase chain reaction was employed to determine TP53INP2 messenger RNA expression in vitro. Western blot analysis was conducted to detect TP53INP2, epithelial-tomesenchymal transition (EMT) marker, and NF-kB/Nrf2/HO-1 pathway protein. The proliferative potentials of glioma cells were assessed by 5-ethynyl-2'-deoxyuridine, colony formation, and cell counting kit-8 assays. Transwell assays were used to evaluate migratory and invasive capacities. Apoptotic cells and reactive oxygen species were analyzed using flow cytometer. Enzyme-linked immunosorbent assay was performed to measure superoxide dismutase and glutathione peroxidase levels. A tumor xenograft model in mouse was established.

Results: High expression of TP53INP2 was observed in glioma cells. TP53INP2 depletion significantly inhibited tumor growth, metastasis, EMT, and oxidative stress and increased the apoptosis rate and number of immune cells. The silenced TP53INP2 hampered the activation of NF-kB and promoted the activation of the Nrf2/HO-1 pathway.

Conclusion: This work highlights the therapeutic potential of TP53INP2/NF-kB/Nrf2/HO-1 axis in SCG.

Objective: Ras p21 protein activator 1 (RASA1) plays a crucial role in the placenta. However, its effects and mechanisms of RASA1 on trophoblast function in preeclampsia (PE) are unclear. This study aims to investigate the relationship between the regulation of the Ras/mitogen-activated protein kinase (MAPK) pathway by RASA1 and the function of trophoblast cells in PE.

Material and methods: Placental tissues were collected from patients with early-onset PE and late-onset PE and their gestational age-matched normal pregnancies. Western blot analysis and quantitative reverse transcription polymerase chain reaction were employed to assess RASA1 levels in placental tissues and trophoblast cells, as well as Ras activation and p38 MAPK phosphorylation levels in the Ras/MAPK pathway. Wound healing, cell counting kit-8, and Transwell migration and invasion assays and flow cytometry experiments were conducted to detect the proliferation, migration, invasion, and apoptosis capabilities of trophoblast cells.

Results: RASA1 was significantly overexpressed in the placental tissues of patients with PE (P < 0.05). In cells, it downregulated the activation of Ras and phosphorylation of p38 MAPK (P < 0.05) and reduced proliferation and inhibited migration and invasive capabilities (P < 0.05). Moreover, RASA1 increased the rate of apoptosis, promoted the protein expression levels of cleaved caspase3 and Bax, and inhibited the expression of Bcl2 in cells (P < 0.05). The P38/MAPK inhibitor SB203580 reversed the activation of the Ras/MAPK pathway and the effects on proliferation, migration, invasion, and apoptosis of cells induced by si-RASA1 (P < 0.05).

Conclusion: The activity of the Ras/MAPK pathway could be inhibited by high RASA1 expression, which suppresses cell invasion, migration, and proliferation and boosts apoptosis. The abnormal regulation of the RASA1-Ras/MAPK axis may be a key factor in the development of PE and, therefore, provides new ideas and clinically effective strategies for the diagnosis and treatment of this condition.

Objective: Patients with chronic kidney disease (CKD) exhibit increased vascular calcification (VC) risks, worsened by high-dose erythropoietin (EPO). While EPO treats anemia, its role in VC pathogenesis remains unclear. Ginsenoside Rb1 (Rb1), a Panax ginseng compound with anti-calcification properties, may counteract EPO-induced VC through the GATA binding protein 6 (GATA6)/bone morphogenetic protein 2 (BMP2)/Smad1/5/9 pathway. This article aims to explore whether Rb1 could counteract EPO-induced VC through the GATA6/BMP2/Smad1/5/9 pathway.

Material and methods: Adenine-induced CKD rats and b-glycerophosphate-treated vascular smooth muscle cells (VSMCs) received EPO ± Rb1. Calcification was assessed through von Kossa/alizarin red staining. Smooth muscle protein 22-a (SM22a)/a-Smooth muscle actin (a-SMA) expression was measured by immunofluorescence and real-time-quantitative polymerase chain reaction (RT-qPCR). GATA6/BMP2/Smad1/5/9 activation was analyzed using RT-qPCR/Western blot. Rb1-BMP2 interactions were tested through biotin pulldown, micro-thermophoresis, and Co-immunoprecipitation (Co-IP). GATA6 knockdown validated pathway roles.

Results: High-dose EPO significantly worsened CKD-associated calcification and VSMC calcification (P < 0.01), suppressed SM22a and a-SMA expression levels, and activated the GATA6/BMP2/Smad1/5/9 pathway (P < 0.01). GATA6 knockdown reduced EPO-exacerbated calcification and modulated BMP2/Smad1/5/9 signaling (P < 0.01). Rb1 increased SM22a and a-SMA expression levels and inhibited Smad 1/5/9 phosphorylation (P < 0.01), without affecting GATA6 or BMP2 expression (P > 0.05). Molecular docking and Co-IP experiments revealed that Rb1 binds directly to BMP2, blocking its interaction with bone morphogenetic protein receptor and inhibiting Smad 1/5/9 phosphorylation (P < 0.01).

Conclusion: Rb1 mitigates EPO-aggravated VC in CKD by disrupting BMP2/Smad1/5/9 signaling, positioning it as a promising molecular intervention strategy to reduce EPO-induced vascular toxicity.

Low-grade fetal adenocarcinoma (L-FLAC) is an exceedingly rare subtype of lung adenocarcinoma, accounting for merely 0.3% of all pulmonary adenocarcinomas. Its accurate intraoperative cytological diagnosis poses substantial challenges, particularly in distinguishing it from carcinoid tumors, which have overlapping morphological features but different therapeutic approaches. We present the case of a 47-year-old female non-smoker with an incidental 21-mm pulmonary nodule spanning the right upper and middle lobes. Intraoperative fine-needle aspiration cytology demonstrated features suggestive of an atypical carcinoid tumor, primarily due to the presence of planar atypical cells with neuroendocrine-like characteristics. Comprehensive cytological assessment revealed two distinct cellular patterns: loosely cohesive planar cells alongside densely nucleated, three-dimensional cohesive clusters with cribriform architecture resembling endometrial tissue. Most importantly, focal cell aggregates with ground-glass nuclei and intranuclear inclusions, corresponding to morule-like structures - a pathognomonic feature of L-FLAC - were identified during detailed examination. Histopathological evaluation confirmed L-FLAC, characterized by atypical glandular proliferation with clear cytoplasm, subnuclear vacuolation, and distinctive morule-like structures demonstrating strong nuclear b-catenin positivity. The patient underwent lobectomy with lymph node dissection and remained recurrence-free at the 2-year follow-up. This case highlights four critical cytomorphological features essential for accurate intraoperative diagnosis of L-FLAC: (1) recognition of dual cell populations (loose planar cells versus cohesive endometrial-like clusters), which contrasts with the monomorphic presentation of carcinoid tumors; (2) identification of ground-glass nuclei and intranuclear inclusions in morule-like structures, features absent in carcinoid tumors; (3) presence of cribriform patterns within cohesive clusters; and (4) awareness that neuroendocrine-like features can dominate the cytological presentation. Accurate distinction between these entities is crucial, as carcinoid tumors may be amenable to limited resection in selected cases, whereas adenocarcinomas generally warrant lobectomy with lymph node dissection. Cytopathologists should remain vigilant for the subtle but diagnostic features of L-FLAC, particularly ground-glass nuclei and intranuclear inclusions, which provide definitive evidence for differentiating this entity from carcinoid tumors in challenging intraoperative settings.

Objective: Ovarian cancer is a disease that seriously endangers the health and life safety of women. At present, no effective preventive and therapeutic measures are available. This study probed the impact of cell division cycle-associated 2 (CDCA2) on ovarian cancer development and cisplatin sensitivity, which provides a new research direction for the study of ovarian cancer.

Material and methods: The protein expression level of CDCA2 was tested by Western blot assay. Cell proliferation was evaluated by cell cloning formation assay and Celigo cell counting. Cell invasion and migration were assessed by Transwell assay. An experiment for nude mouse tumor formation was conducted to analyze the influence of CDCA2 knockdown on tumor growth in vivo. We treated CDCA2 knockdown cells with gradient cisplatin and measured cell viability using the cell counting kit-8 assay. Apoptosis and DNA damage induced by CDCA2 knockdown were investigated by flow cytometry and histone family member X (H2AX) phosphorylated on Ser 139 (γ-H2AX) immunofluorescence, respectively.

Results: CDCA2 expression was knocked down in A2780 and SKOV3 cells. After CDCA2 knockdown, cell proliferation, migration, and invasion ability decreased significantly, and tumor growth in vivo was also limited (P < 0.01). The phosphorylation levels of protein kinase B (AKT) and mechanistic target of rapamycin (mTOR) were reduced by CDCA2 knockdown (P < 0.01), but the effect was reversed by the AKT activator SC-79 (P < 0.01). Knockdown of CDCA2 increased the cisplatin sensitivity of ovarian cancer cells by enhancing apoptosis and DNA damage (P < 0.01).

Conclusion: CDCA2 knockdown inhibited the development of ovarian cancer through the AKT/mTOR pathway and enhanced cisplatin sensitivity. CDCA2 is a potential target to reverse cisplatin resistance in ovarian cancer. It can also be used as a new research direction for the development of ovarian cancer therapy.

Objective: The objectives of the study are to investigate the differential metabolic paradigms and distribution of regulatory T (Tregs) cells between primary prostate cancer (PCa) and lymph node (LN) metastases.

Material and methods: Single-cell RNA sequencing analysis of primary PCa and LN metastases was employed to reveal the immune infiltration, identify Treg cell clusters, and analyze their metabolic regulation. Immunohistochemical (IHC) for FOXP3 and cluster of differentiation antigen 45 was used to verify different distribution and infiltration of Treg cells.

Results: Immune cell infiltration was prominent around PCa cells, with Tregs significantly enriched in node-positive samples, suggesting an immunosuppressive microenvironment. Three Treg subsets were identified: Inhibitory Tregs, effector Tregs, and double-positive Tregs, each exhibiting distinct metabolic profiles. IHC confirmed higher Treg infiltration in LN metastases compared to primary tumors, particularly within tumor stroma.

Conclusion: Tregs promote lymphatic metastasis in PCa through metabolic reprogramming, with their infiltration levels serving as a potential biomarker for metastatic risk.