Cytojournal最新文献

Objective: Pregnancy-induced hypertension (PIH) is a common complication during pregnancy and is closely associated with vascular endothelial cell damage and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)-mediated pyroptosis. This study aimed to investigate whether mitophagy alleviates vascular endothelial cell damage in PIH by inhibiting NLRP3-mediated pyroptosis. The regulatory mechanisms of pyroptosis-related pathways were systematically investigated by establishing a cellular model of PIH and incorporating mitophagy intervention.

Material and methods: An Nω-nitro-L-arginine methyl ester (L-NAME)-induced gestational hypertension model was established, and the cell samples were grouped as follows: Control group (Control), L-NAME-induced gestational hypertension group (L-NAME), mitochondrial autophagy inhibition group (L-NAME+ 3-methyladenine [3-MA]), and mitochondrial autophagy activation group (L-NAME+ rapamycin [Rapa]). Cell viability was assessed through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, lactate dehydrogenase (LDH) levels were measured to evaluate cell damage, and reactive oxygen species (ROS) kits were used to quantify ROS accumulation. Cell death was evaluated using terminal deoxynucleotidyl transferase dUTP nick end labeling staining to detect apoptotic cells. Immunofluorescence, Western blot analysis, and quantitative real-time polymerase chain reaction were performed to assess the expression levels of proteins and genes associated with mitophagy (e.g., microtubule-associated protein 1 light chain 3 and sequestosome 1) and those linked to pyroptosis (e.g., NLRP3, gasdermin D (GSDMD), cysteinyl aspartate-specific proteinase 1 (caspase-1), interleukin (IL)-1β, and IL-18). The role of NLRP3 in pyroptosis regulation through mitochondrial autophagy was further examined using NLRP3 small interfering RNA (siNLRP3) transfection experiments.

Results: L-NAME treatment substantially decreased vascular endothelial cell viability, elevated LDH release and ROS levels, and upregulated pyroptosis-related proteins (NLRP3, GSDMD, and caspase-1) and inflammatory factors (IL-1β and IL-18). The inhibition of mitochondrial autophagy with 3-MA further enhanced pyroptosis and aggravated cell damage, and its activation with Rapa reduced pyroptosis, improved cell survival, and decreased LDH release and ROS levels. NLRP3 silencing (siNLRP3) significantly inhibited pyroptosis and alleviated the cell damage caused by 3-MA. Meanwhile, Rapa enhanced the protective effect of NLRP3 silencing.

Conclusion: This study demonstrates that mitophagy can effectively alleviate the vascular endothelial cell damage associated with PIH by inhibiting NLRP3-mediated pyroptosis. The findings provide new theoretical support for the treatment of PIH and suggest potential intervention targets.

Objective: Histopathology examination is important for diagnosing autoimmune pancreatitis (AIP), which is suspected to be pancreatic cancer based on imaging findings. Although the validity of endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) in the diagnosis of AIP is still debated globally, this study aimed to evaluate the efficacy of EUS-FNA in the diagnosis of AIP with suspected pancreatic cancer.

Material and methods: From January 2021 to June 2024, 30 AIP patients with radiographically diagnosed pancreatic cancer were enrolled and underwent EUS-FNA. Sex, age, symptoms, CA199, serum immunoglobulin G4 (IgG4), and treatment outcome were included. Tissue sampling conditions, puncture sites, storiform fibrosis, CD38- and IgG4-positive plasma cell counts, and obliterans phlebitis were evaluated.

Results: Thirty patients, 24 males and six females, with an average age of 60.53 ± 11.72 years (32-79 years), were included in the study. Thirty patients had their serum IgG4 and CA199 levels tested. Tissue samples containing ≥10 were obtained from 19 (63.33%) patients. CD38+ plasma cell infiltration and laminar fibrosis were detected in 22 (73.33%) and 10 (33.33%) patients. According to the International Consensus Diagnostic Criteria ( ICDC), 12 patients had histopathological levels of Grade 1, 15 of Grade 2, and three patients could not be classified. The accuracy, sensitivity, and specificity of EUS-FNA in diagnosing AIP with suspected pancreatic cancer on imaging were 96.66% (29/30), 96.42% (27/28), and 100% (2/2), respectively. The area under the curve value of EUS-FNA for patients with AIP who were radiologically suspected of having pancreatic cancer was 0.957.

Conclusion: Approximately 90% of patients with EUS-FNA results are diagnosed with an ICDC level of 2 or higher. Our results suggest that for cases where malignant tumors are suspected after imaging or cannot be ruled out, obtaining pancreatic tissue through EUS-FNA puncture for pathological diagnosis is recommended.

Objective: This study examined the role of sex-determining region Y-box 4 (SOX4) in sorafenib-resistant hepatocellular carcinoma (HCC) cells and its potential therapeutic relevance by focusing on the effects of SOX4 knockdown on tumor growth, apoptosis, and immune infiltration.

Material and methods: A sorafenib-resistant HCC cell line (sorafenib-resistant HepG2 [SR-HepG2]) was established by gradually increasing the sorafenib dose (1-7 μM) over 12 months. The messenger RNA and protein expression levels of SOX4 in HepG2 and SR-HepG2 cells were analyzed by a quantitative reverse transcription-polymerase chain reaction and Western blot. Small interfering RNA (SOX4) or SOX4 overexpression plasmids were introduced into SR-HepG2 cells through transfection, and the effects on cell proliferation, colony formation, and apoptosis were evaluated using 5-ethynyl-2'-deoxyuridine staining, colony formation assays, and terminal deoxynucleotidyl transferase dUTP nick end labeling assays. For in vivo experiments, HepG2 or SR-HepG2 cells were subcutaneously injected into BALB/c nude mice to monitor tumor growth. In the sorafenib-resistant HCC mouse model, SOX4 knockdown (small-interfering RNA SOX4 [si-SOX4]) was delivered through lentiviral vectors to assess its effect on tumor growth. Immune cell infiltration was assessed by immunofluorescence staining, and the influences on immune escape markers were evaluated by Western blot.

Results: Compared with those in the parental HepG2 cells, the transcriptional and translational expression levels of SOX4 were significantly elevated in the SR-HepG2 cells (P < 0.001). Si-SOX4 markedly suppressed the proliferation and colony formation of SR-HepG2 cells and increased their cell apoptosis (P < 0.001). In vivo experiments revealed that si-SOX4 inhibited tumor growth in the sorafenib-resistant HCC model, accompanied by a significant reduction in tumor volume and weight (P < 0.001). Histological analysis showed that si-SOX4 disrupted the tumor structure, characterized by increased necrosis and reduced collagen fibers. In addition, si-SOX4 decreased the infiltration of Forkhead box P3+regulatory T cells and cluster of differentiation 11b + myeloid-derived suppressor cells while increasing the number of cluster of differentiation 8 (CD8)+ T cells and granzyme B + CD8+ cytotoxic T cells (P < 0.001). SOX4 knockdown also reduced the expression of two immune escape markers, programmed cell death ligand 1 and C-C motif chemokine ligand 12 (P < 0.001).

Conclusions: SOX4 overexpression drives sorafenib resistance in HCC cells by promoting cellular growth, inhibiting apoptosis, and enhancing immune evasion. Conversely, SOX4 knockdown inhibits tumor growth, alters immune cell infiltration, and reduces immune escape. Hence, targeting SOX4 is a promising therapeutic approach to overcome sorafenib resistan

Objective: In recent years, several publications have described the use of ultrasound-guided fine-needle aspiration (FNA) by cytopathologists to achieve better diagnostic accuracy. Some cytopathologists enroll in courses to learn and apply ultrasound (US) guidance themselves. However, no standard procedure has been established that cytopathologists can follow to perform US for FNA. Alternatively, FNA can be a useful tool when cytopathologists collaborate with radiologists. Here, we aimed to evaluate the diagnostic accuracy of FNA for non-thyroidal head-and-neck masses retrieved by a cytopathologist with US guidance provided by a radiologist.

Material and methods: The FNA results for non-thyroidal head-and-neck masses at a private clinic using the Scandinavian FNA model with radiologist‒cytopathologist collaboration were compared with the histopathology results.

Results: In all, 1890 patients who underwent FNA were identified, among whom 1435 (76%) also had histopathological results. Non-cystic lesions were obtained from lymph nodes (LNs), salivary glands, and soft tissue, while the other lesions were cystic in nature. For FNA, the accuracy was 99.4%, the sensitivity was 99.6%, the specificity was 99.3%, the positive predictive value was 99.3%, and the negative predictive value was 99.6%. No FNA results were non-diagnostic. Surgical follow-up revealed that only eight of the 1435 assessments (0.5%), all performed for LN lesions, yielded false-negative or false-positive results.

Conclusion: The present study is based on single-center observations. The use of FNA, when performed by a specialized cytopathologist and with US assistance from a radiologist, produces accurate results and sufficient material for analysis, especially for LNs in extrathyroidal head-and-neck lesions. This study also reveals that the technique is a low-cost and effective process. The way in which FNA is presented here indicates that this procedure would be useful and ideal for any health service.

Objective: The Japanese System for Reporting Thyroid Cytopathology (JSRTC) does not include the risks of malignancies (ROMs) or recommended clinical management. This multi-institutional study aimed to determine the frequency, re-aspiration rate, resection rate, ROM, and clinical management options in seven different categories.

Material and methods: For 15,495 cases of thyroid fine-needle aspiration performed at seven Japanese institutions without molecular testing, the frequency, re-aspiration rate, resection rate, ROM, and clinical management options of each diagnostic category were examined. The categorization was based on JSRTC, and cases were subdivided into those with nuclear atypia and other subtypes for undetermined significance.

Results: Re-aspiration of unsatisfactory and undermined significance diagnostic categories was mainly performed for cases of suspected malignancy on ultrasound. The median re-aspiration rate of cyst fluid nodules was 4.9%, which was significantly different from that (17.8%) of unsatisfactory cases (P < 0.05). The resected ROMs for nodules that were suspicious for malignancy and malignant were 94.2% and 99.6%, respectively. The low resection rates of nodules that were suspicious for malignancy (77.8%) and malignant (70.8%) could be attributed to active surveillance for low-risk papillary microcarcinoma. The overall ROMs of unsatisfactory, cyst fluid, benign, undetermined significance, and follicular neoplasms were 4.5%, 0.4%, 0.7%, 16.7%, and 11.4%, respectively. In the subtype of undetermined significance, the overall ROM of nuclear atypia (27.6%) was higher than that of the others (6.7%).

Conclusion: Overall, this study determines the frequency, ROM, and recommended clinical management for thyroid cytopathology in Japan. These results were different from those proposed by the Bethesda System for Reporting Thyroid Cytopathology. In the future, our results will be helpful in the revision of JSRTC and will contribute to improving the outcomes among Japanese patients with thyroid nodules.

Objective: Endometrial cancer (EC) ranks among the most prevalent malignant tumors affecting women, with metastasis and dissemination as the major contributors to poor prognosis. This study explores the involvement of forkhead box protein P2 (FOXP2) in EC cell invasion and migration, which is mediated through the activation of myosin light-chain kinase (MYLK).

Material and methods: Bioinformatic analysis was conducted to determine whether FOXP2 is expressed in EC. FOXP2 overexpression was achieved using a FOXP2 overexpression vector (oeFOXP2), and negative control (NC) was used for cell transfection. Cell counting kit-8, 5-ethynyl-2'-deoxyuridine, enzyme-linked immunosorbent assay, colony formation, wound healing, and Transwell assay were used to assess the capabilities of cell viability, invasion, migration, and proliferation. Western blot and quantitative real-time polymerase chain reaction (qRTPCR) analysis were used to measure the expression levels of proteins linked to the epithelial-mesenchymal transition. The correlation between FOXP2 and MYLK was analyzed using bioinformatics and validated by Western blot and qRT-PCR analysis. The MYLK-specific inhibitor ML-7 was employed to study the impact of MYLK-mediated FOXP2 on regulating the malignant biological processes of EC.

Results: The oeFOXP2 group of EC cells exhibited a significant decrease in cell viability, colony formation, migration rate, and metastatic cell count compared with the NC group (P < 0.05). FOXP2 overexpression markedly increased caspase-3, caspase-8, caspase-9 activity (P < 0.05). Significant changes were detected in the expression of epithelial-mesenchymal transition marker proteins, with vimentin and N-cadherin expression noticeably declining and E-cadherin expression sharply rising (P < 0.05). The addition of the MYLK-specific inhibitor ML-7 reversed the effect of FOXP2 overexpression on the invasion and migration of EC cells.

Conclusion: FOXP2 suppresses the proliferation, invasion, and migration of EC cells through the activation of MYLK.

Objective: Concurrent chemotherapy and radiotherapy (CCRT) has been applied as a therapeutic modality for cervical squamous cell carcinoma (CESC). Our aim is to investigate the potential marker(s) of the efficacy of CCRT in CESC.

Material and methods: Potential candidates predictive of the efficacy of CCRT in CESC were identified. Differentially expressed genes (DEGs) were screened, followed by performing functional enrichment analyses. CCRT-related biomarkers were identified. In addition, the CIBERSORT algorithm was employed to determine the immune cell infiltration. Immune cell subsets from donors and specific cytokines were evaluated, and the biological functions of CESC cells following cisplatin treatment or coculture with M2 macrophages were explored.

Results: A total of 56 DEGs were singled out. These DEGs were enriched in pathways relevant to CESC and CCRT. They were narrowed down to eight CCRT-related biomarkers with good predictive values. Notably, most of the biomarkers were negatively correlated with M2 macrophages (P < 0.05), and regulator of G-protein signaling 2 (RGS2) exhibited low expression in CESC (P < 0.05). Flow cytometry results revealed that patients with CCRT-resistant CESC had high percentages of M2 macrophages, CD4 T cells, regulatory T cells and T helper 2 cells but low percentages of T helper 1 cells, and T helper 17 cells, M1 macrophages, and CD8 T cells (P < 0.05). Aside from interleukin (IL4) and IL-10, the remaining specific cytokines exhibited low expression in patients with CCRT-resistant CESC (P < 0.05). Furthermore, the cell cycle progression and metastasis of CESC cells were evidently promoted by M2 macrophages but were suppressed by cisplatin intervention (P < 0.05). Moreover, in CESC cells, cisplatin repressed the levels of IL-4 and IL-10 yet boosted those of the remaining cytokines, whereas M2 macrophages had the opposite effects (P < 0.05). RGS2 silencing promoted the phosphorylation of phosphatidylinositol 3-kinase/protein kinase B/transcriptional signal transducer and activator 6 in macrophages, whereas RGS2 overexpression had the opposite effect (P < 0.05).

Conclusion: This study interpreted and explored the possible predictive values of RGS2 in the efficacy of CCRT in CESC. It may provide other insights for the management of CESC.

Objective: PDZ domain containing 1-interacting protein 1 (PDZK1IP1) is commonly overexpressed in a wide variety of cancer. Hence, the objective of the present study is to ascertain the influences of PDZK1IP1 on colorectal carcinoma (CRC) development.

Material and methods: PDZK1IP1 expression was tested through reverse transcription-quantitative polymerase chain reaction and Western blot analysis, and its correlation with prognosis was analyzed using the GEPIA website. Small interfering RNA against PDZK1IP1 was adopted to downregulate PDZK1IP1 expression in CRC cells. The effects of PDZK1IP1 on cell growth were ascertained using colony formation and CCK-8 tests, and CRC cell apoptosis was analyzed through flow cytometry. Cell migration capability and invasiveness were measured using Matrigel Transwell and scratch-healing assays.

Results: PDZK1IP1 was highly expressed in the CRC tissues (P < 0.001) and cells (P < 0.05), and its knockdown restrained cell growth (P < 0.05), migratory potential (P < 0.01), and invasive capacities (P < 0.001) and accelerated cell apoptosis (P < 0.001). Mechanically, PDZK1IP1 silencing blocked CRC progression by inactivating the phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of the rapamycin pathway.

Conclusion: PDZK1IP1 contributes to the oncogenesis of CRC. This finding provides a basis for the diagnosis, treatment, and prevention of CRC.

Objective: This study aims to investigate the role of fat mass and obesity-associated protein (FTO) in renal clear cell carcinoma (RCC), particularly its regulatory effects on glycolysis, cell proliferation, and sirtuin 1/forkhead box O1 (SIRT1/FOXO1) signaling pathway.

Material and methods: The messenger RNA and protein expression levels of FTO in human proximal tubular epithelial cells (human kidney 2 [HK-2]) and the RCC cell line A498 were determined by quantitative reverse transcription polymerase chain reaction and Western blot. FTO expression was downregulated by FTO short hairpin RNA and overexpressed using plasmids. Glycolysis levels were assessed by measuring glucose uptake, lactate secretion, extracellular acidification rate, and adenosine triphosphate/adenosine diphosphate (ATP/ADP) ratio. The effects of FTO on cell proliferation and cell cycle were evaluated through colony formation assays, 5-ethynyl-2'-deoxyuridine (EdU) staining, and flow cytometry. The SIRT1/FOXO1 signaling pathway was analyzed through Western blot, and FOXO1 pathway inhibitor (AS1842856) was used to further explore the role of SIRT1/FOXO1 in the FTO-mediated regulation of RCC.

Results: FTO was downregulated in A498 cells compared with that in HK-2 cells. FTO downregulation markedly increased glucose uptake, lactate secretion, and the ATP/ADP ratio in A498 cells, and its overexpression inhibited these processes. FTO downregulation also promoted RCC cell proliferation, as evidenced by an increase in colony formation and the number of EdU-positive cells. Meanwhile, FTO overexpression suppressed the proliferation of these cells. Flow cytometry analysis revealed that FTO downregulation notably increased the proportion of cells in the S phase, and its overexpression increased the proportion of cells in the G0/G1 phase. Further analysis indicated that FTO downregulation activated the SIRT1/FOXO1 signaling pathway, and its overexpression inhibited this pathway. Treatment with the FOXO1 inhibitor AS1842856 significantly reversed the pro-glycolysis and pro-proliferation effects of FTO downregulation, supporting the role of the SIRT1/FOXO1 pathway in FTO-mediated regulation.

Conclusion: FTO downregulation promotes glycolysis and proliferation in RCC cells by activating the SIRT1/ FOXO1 signaling pathway. Targeting the FTO and SIRT1/FOXO1 pathway may provide potential therapeutic strategies for the treatment of RCC.

Objective: Pancreatic cancer is a major global health challenge with high mortality rates and limited therapeutic options. Fine-needle aspiration (FNA) cytology is a key diagnostic tool, but discrepancies between cytological and histological diagnoses can impact patient management. This study aims to evaluate the diagnostic accuracy of pancreatic FNA using the World Health Organization (WHO) reporting system to assess the risk of malignancy (ROM) across different diagnostic categories.

Material and methods: The WHO reporting system was employed to reclassify 122 FNAs, with 37 cases undergoing subsequent histological correlation to evaluate the ROM. The sensitivity, specificity, positive and negative predictive values, and accuracy of ROM using the WHO system were determined through statistical analyses.

Results: The discrepancy rate between cytology and histology diagnoses was 16.2%. Category 6 (malignant) showed consistent ROM values (89%), confirming its reliability in predicting malignancy. However, Categories 1, 2, and 3 had higher ROM values than previously reported, while Category 4 had a lower ROM. Factors such as small lesion size, poor cellularity, and sampling limitations contributed to diagnostic discrepancies.

Conclusion: The study offers significant insights into the cyto-histopathological correlation in pancreatic FNA, highlighting the effectiveness of the WHO reporting system in ROM assessment. Future research with larger samples is necessary to enhance the accuracy of pancreatic FNA cytology for improved patient outcomes.