Cytojournal最新文献

Objective: Transmembrane Emp24 Domain Containing 2 (TMED2) is a mediator of membrane protein trafficking involved in intracellular protein transport. Recent research suggests that TMED2 plays an important role in the development and metastasis of tumors; however, its exact mechanisms in oral cancer (OC) remain unclear. This study aims to elucidate the role and possible mechanisms of TMED2 in OC.

Material and methods: We investigated the impact of TMED2 knockdown on the invasion, migration, and proliferation capabilities of OC cells. Furthermore, we analyzed the in vitro and in vivo interactions between TMED2 and polypeptide-N-acetylgalactosaminyltransferase 7 (GALNT7) as well as explored the regulatory function of TMED2 on GALNT7. The alterations in stem cell markers were assessed using clone formation assays, western blot, and quantitative real-time polymerase chain reaction.

Results: The upregulation of TMED2 promoted the proliferation and invasion abilities of OC cells. Further analysis revealed that TMED2 enhanced the stem-like properties and tumorigenicity of OC cells by directly regulating the expression of GALNT7. In vivo and in vitro results suggested that silencing TMED2 expression reduced the incidence of OC.

Conclusion: Our data imply that TMED2 stimulates GALNT7 transcription, which in turn amplifies the stem-like characteristics and carcinogenic potential of OC cells. Moreover, the block of TMED2 prevents cancers from growing and spreading in vivo. This finding provides a new therapeutic target for the treatment of OC and highlights the critical role of TMED2 in the condition.

Objective: Soft tissue and bone cancers, collectively known as sarcomas, constitute a diverse array of uncommon tumors originating from connective tissues. Among sarcomas, leiomyosarcoma (LMS) is one of the most frequently encountered subtypes. This study aims to investigate the expression, clinical significance, biological regulation, and dysregulation mechanisms of extra spindle pole bodies like 1 (ESPL1), a gene critical for cell cycle regulation in LMS.

Material and methods: Bioinformatics analysis was performed using the data from The Cancer Genome Atlas-Sarcoma and Genotype-Tissue Expression datasets. Functional experiments to assess cell proliferation and the cell cycle were performed in LMS cells (SK-LMS-1) after ESPL1 knockdown. Bioinformatics analyses were conducted to identify the potential transcriptional regulators of ESPL1. The regulatory relationship between ESPL1 and the E2F transcription factor 1 (E2F1) was validated through the various molecular assays.

Results: ESPL1 is significantly overexpressed in LMS compared with normal muscle tissue. High ESPL1 expression is associated with a shorter progression-free interval (PFI) in sarcoma patients, particularly in the LMS subset. ESPL1 expression might be an independent prognostic factor for poor overall survival and PFI in LMS patients. Functional studies in the LMS cell line SK-LMS-1 demonstrated that ESPL1 knockdown slowed cell proliferation and increased G2/M cell cycle arrest, suggesting its crucial role in maintaining LMS cell viability and genomic integrity. Further bioinformatics analysis identified the E2F1 transcription factor as a key regulator of ESPL1 expression in LMS. Mechanistic investigations demonstrated that E2F1 interacts with the ESPL1 promoter, leading to transcriptional activation.

Conclusion: These findings highlight the ESPL1-E2F1 axis as a potential prognostic biomarker and therapeutic target in LMS.

Objective: Microglial activation is a hallmark of pathogenic retinal conditions such as retinal ischemia-reperfusion (RIR). While sortilin-related vacuolar protein sorting 10 domain containing receptor 2 (Sorcs2) and laminin subunit alpha 1 (Lama1) have been implicated in neuroinflammatory processes, their roles in regulating microglial activation in RIR are not reported. The current work studied the potential of Sorcs2 and Lama1 as negative regulators of microglial activation in RIR and assessed the therapeutic potential of Astragalus polysaccharide (AP).

Material and methods: Transcriptome profiling was conducted in retinal specimens of RIR group 72 h after RIR induction. Oxygen-glucose deprivation/reperfusion (OGD/R) in rat microglial cells was employed as the cellular induction model of RIR. The functional role of Sorcs2 and Lama1 in dictating microglial activation was investigated in vitro and in vivo using lentivirus-based gene expression. Further, the potential effect of AP on RIR-mediated microglial activation was investigated.

Results: Sorcs2 and Lama1 were identified as two downregulated genes in retinal samples following RIR. OGD/R induction triggered pro-inflammatory microglial activation and induced the downregulation of Sorcs2 and Lama1. Sorcs2 or Lama1 overexpression hindered OGD/R-induced microglial activation in vitro and attenuated inflammatory expansion of microglia cells in RIR-induced rat retinal samples. AP treatment was able to neutralize the oxidative stress, promote the expression of Sorcs2 and Lama1, and suppress microglial activation.

Conclusion: Our findings pinpoint Sorcs2 and Lama1 as negative regulators of microglial activation in RIR. AP could be employed as an antioxidant to attenuate microglial activation and ameliorate the inflammatory damages in RIR.

Objective: The conflicting results of the Bethesda system for reporting thyroid cytopathology (BSRTC) and B-Raf proto-oncogene (BRAF) mutation status during pre-operative fine-needle aspiration cytology (FNAC) of thyroid nodules create a dilemma for clinicians in devising appropriate treatment strategies for patients. This study provides a report on the histopathological findings of 687 thyroid nodules with an indeterminate cytological diagnosis after the combination of the BSRTC and BRAF mutation status.

Material and methods: The clinical data of patients with thyroid nodules, suspicious of malignancy at ultrasound (US), who underwent US-guided FNAC between December 2020 and March 2023 at our cancer center were reviewed. Patients with an indeterminate diagnosis, that is, conflicting results of the BSRTC and BRAF mutation status after FNAC, were enrolled. The following four combinations of BSRTC and BRAF mutation status were considered indeterminate: (1) Group 1, BSRTC I and positive for a BRAF mutation; (2) Group 2, BSRTC II and positive for a BRAF mutation; (3) Group 3, BSRTC III and positive for a BRAF mutation; and (4) Group 4, BSRTC V and negative for a BRAF mutation. Finally, only patients who underwent surgical treatment at our center were included in the data analysis.

Results: Among the 1,044 eligible patients, 687 underwent surgical treatment. Of the 687 patients, 117 were in Group 1, 14 in Group 2, 394 in Group 3, and 162 in Group 4. Histopathological examination showed that 677 (98.5%) patients had papillary thyroid cancer, including 585 with papillary thyroid microcarcinoma, whereas only 10 (1.5%) had benign nodules. The malignancy rates were 98.3%, 100%, 98.7%, and 98.1% for Groups 1 to 4, respectively. Among the 387 patients in category 4A by the thyroid imaging reporting and data system (TI-RADS 4A) through the US, the malignancy rate was 98.4%, and for the 116 nodules <5 mm in diameter in the US, the malignancy rate was 99.1%. When combining TI-RADS 4A and a nodule diameter <5 mm, the malignancy rate was 98.9% (88/89). A total of 179 patients (26.1%) had histopathologically confirmed central cervical lymph node metastasis, and 46 (6.8%) had lateral cervical lymph node metastasis. Two nodules in Group 1, five nodules in Group 3, and three nodules in Group 4 were determined to be benign post-surgery. The benign thyroid nodules included seven dysplastic, one adenomatous, one fibrotic, and one hyperplastic.

Conclusion: Thyroid nodules, suspicious of malignancy on US, after the combined interpretation of BSRTC and BRAF mutation status following pre-operative FNAC had a high risk of malignancy. Repeat US-guided FNAC for indeterminate thyroid nodules is highly recommended in clinical practice.

Objective: Crohn's disease (CD) is a chronic inflammatory condition of the bowel that remarkably impairs a patient's quality of life and often has a poor prognosis. Perianal fistulizing CD (PFCD) is one of the most common parenteral symptoms of CD and a huge challenge for the management of this illness. This study aimed to elucidate the molecular mechanisms underlying PFCD and identify potential biomarkers to advance our understanding and management of this condition.

Material and methods: Transcriptome sequencing was performed using the control and PFCD groups to investigate the mechanisms of PFCD development. The expression of tumor necrosis factor receptor-associated factor 5 (TRAF5), nuclear factor-kappa B (NF-κB), and interleukin 13 (IL-13) messenger ribonucleic acid (mRNAs) was detected by quantitative polymerase chain reaction (qPCR). Pathological morphology was observed using hematoxylin and eosin staining. The expression of TRAF5, Epithelial Cadherin (E-cadherin), Snail family transcriptional repressor 1 (SNAIL1), and vimentin protein was detected by immunohistochemistry. Following the knockdown of TRAF5 in human tumor-29 (HT-29) cells, the effects on cell proliferation and migration were assessed using the cell counting kit-8 and Transwell assays. The expression levels of crucial markers were analyzed by qPCR, Western blot, and immunohistochemistry.

Results: Transcriptomic sequencing revealed a significant upregulation of TRAF5 in the PFCD group, accompanied by elevated mRNA levels of NF-κB and IL-13 compared with those in the control group. In addition, the PFCD group exhibited increased expression of TRAF5, SNAIL, and vimentin and marked reduction in E-cadherin levels, indicating that PFCD may facilitate epithelial-mesenchymal transition (EMT). Knocking down TRAF5 in HT-29 cells reduced cell proliferation and migration; inhibited NF-κB and IL-13 mRNAs, SNAIL1, and vimentin levels; and promoted E-cadherin levels.

Conclusions: The development of PFCD was associated with EMT, and TRAF5 was a key gene of PFCD. Knocking down TRAF5 alleviated the EMT promotion of PFCD, indicating that TRAF5 drove the development of PFCD through EMT.

Objective: Intrahepatic cholangiolithiasis (Intrahepatic bile duct stones, IBDSs) is a common hepatobiliary disease characterized by bile duct obstruction and inflammation, often leading to severe complications such as cholangitis, cirrhosis, and cholangiocarcinoma. This study investigates the role of fat mass and obesity-associated (FTO) protein, an RNA demethylase, in regulating Kupffer cell (KC) polarization, interleukin (IL)-6 secretion, and subsequent human intrahepatic biliary epithelial cell (HiBEC) proliferation in IBDS.

Material and methods: Liver tissues from patients with IBDS were analyzed for FTO expression, KC M2 polarization, and IL-6 levels. In vitro experiments with FTO silencing in KCs were conducted to examine the effects on M2 polarization, IL-6 production, and HiBEC proliferation. Mechanistic analysis focused on the c-Jun N-terminal kinase (JNK)/p38 and phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) pathways.

Results: The patients with IBDS showed significantly higher KC M2 polarization, elevated FTO expression, and increased IL-6 levels relative to the controls. Without FTO silencing, IL-6 secretion and HiBEC proliferation remained at high baseline levels. However, FTO silencing reduced M2 polarization, IL-6 secretion, and HiBEC proliferation through the JNK/p38 pathway. Activating the PI3K/AKT pathway partially reversed these inhibitory effects.

Conclusion: FTO plays a critical role in IBDS by promoting the M2 polarization of KCs, which leads to increased IL-6 secretion and induced pathological HiBEC proliferation. Targeting FTO may represent a novel therapeutic strategy for managing IBDS and preventing disease progression.

Objective: Sputum cytology is recognized as a straightforward and noninvasive way to diagnose lung cancer, although its clinical utility has not yet been investigated. The objective of the study was to detect and classify cancerous cells in sputum by examining their expression of minichromosome maintenance proteins (MCM2 and MCM7). In addition, the study attempted to evaluate these proteins' potential as biomarkers of lung cancer lesions and their relationships with clinicopathological characteristics.

Material and methods: MCM2 and MCM7 expression in sputum samples was evaluated using immunocytochemistry in sputum cell blocks (n = 97), and their correlation with clinicopathological features was examined. Diagnostic performance was evaluated as a function of sensitivity and specificity.

Results: Immunoexpression of MCM2 and MCM7 was confined to the nuclei of malignant cells alone, suggesting its potential as a differential diagnostic marker. They showed significant correlations with tumor cytology (P < 0.001), while MCM7 alone exhibited a significant correlation with tumor stage (P = 0.014). The overexpression of these markers was notably pronounced in lung adenocarcinoma compared to other subtypes. In terms of characterizing malignant cells, MCM7 protein demonstrated the highest sensitivity at 92% with an area under the curve (AUC) of 0.961, whereas MCM2 had a sensitivity of 80% and AUC of 0.901.

Conclusion: This study presents the inaugural use of MCM7 immunocytochemistry on exfoliated cells in sputum samples, proposing that analyzing immunocytochemical markers in sputum could serve as a cost-effective approach for diagnosing lung cancer. Integrating these assessed markers into routine cytopathology laboratories could augment traditional morphological evaluations, thereby improving the sensitivity of sputum cytology.

Objective: Flow cytometry (FC) can be an adjunct to fine-needle aspiration cytology (FNAC) in the diagnosis and subclassification of hematolymphoid lesions. This study evaluates the utility of FC in diagnosing extramedullary hematolymphoid lesions in fine-needle aspirate samples.

Material and methods: This cross-sectional study enrolled patients who presented to the FNAC clinic and suspected to have hematolymphoid lesions (nodal and extra nodal lesions) from August 2020 to June 2022. Sixty-seven cases of hematolymphoid malignancies and 67 cases without hematolymphoid malignancies were included. The combined FNAC/FC diagnosis was compared with the gold standard: Biopsy, cell block, or peripheral blood/bone marrow aspirate FC.

Results: Of 67 lymphoma cases, 63 were of the primary type and 4 were diagnosed with suspected recurrence/residual disease. Moreover, 57 cases were nodal, and 10 were extranodal. Four of the patients had discordant findings between FNAC and FC. The gold standard was available only in 56 cases, of which 5 had discordant findings between combined FNAC/FC and the gold standard. We subclassified 34 cases based on combined FNAC/FC. The five cases included anaplastic large cell lymphoma (1/5), classic Hodgkin lymphoma (3/5), and one case of atypical lymphoid hyperplasia. Non-contributory FC was attributed to the presence of large cells/necrosis/nodular lymphocyte predominant Hodgkin lymphoma/metastasis/Castleman disease/technical problem.

Conclusion: Combining FC with FNAC enhances diagnostic accuracy and helps subclassify lymphoma. Future works should explore whether it can replace excision biopsy, especially in recurrent cases.

Objective: Geniposide (GP) provides myocardial cells with protection against pyroptosis-induced damage. However, the mechanisms governing GP's effect on the thioredoxin-interacting protein (TXNIP)/nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) signaling pathway remain unclear. This study aimed to explore how GP alleviates post-myocardial infarction (MI)-induced pyroptosis through regulation of the TXNIP/NLRP3 pathway.

Material and methods: In vivo studies: MI models were established, mouse body weight, heart rate, and blood glucose levels were monitored, and methods, such as cardiac ultrasound, hematoxylin-eosin staining, triphenyltetrazolium chloride staining, terminal deoxynucleotidyl transferase 2'-deoxyuridine 5'-triphosphate nick end labeling staining, quantitative polymerase chain reaction (qPCR), and Western blot (WB), were used to explore the effect of GP on myocardial cell pyroptosis. We explored the role of NLRP3 in GP's antimyocardial cell pyroptosis through qPCR, WB, immunofluorescence, enzyme-linked immunosorbent assay (ELISA), and other methods. In vitro studies: A chronic hypoxia (CH) cell model was established, and detection methods, such as cell counting kit-8 assay, transmission electron microscopy, ELISA, and immunological assays, were used to explore the effects of GP on CH myocardial cell pyroptosis and GP's inhibition of the TXNIP/NLRP3 signaling pathway to resist CH myocardial cell pyroptosis.

Results: In vivo studies revealed that after the treatment with GP, the infarct area of mice's hearts significantly decreased, cardiac structure and function notably improved, fibroblast proliferation in cardiac tissues decreased significantly, and the pyroptosis level of myocardial cells decreased. GP treatment significantly downregulated the expression levels of type I collagen (Col I), Col III, TXNIP NLRP3, caspase-1, and gasdermin D N-terminal (GSDMD-N). The inhibition of NLRP3 also reduced the expressions of NLRP3, TXNIP, caspase-1, and GSDMD-N in the cardiac tissue, which is concomitant with a decline in reactive oxygen species (ROS) production. In addition, in vitro studies unveiled that GP effectively alleviated pyroptosis in CH myocardial cells, reducing pyroptosis rates, interleukin (IL)-1β, IL-18, lactate dehydrogenase, and creatine kinase-muscle/brain levels. This protective effect was achieved by inhibiting the TXNIP/NLRP3 signaling pathway.

Conclusion: GP greatly diminishes the extent of infarcted myocardial tissue and mitigates pyroptosis, which improves cardiac structure and function through modulation of the TXNIP/NLRP3 pathway. Furthermore, the inhibition of NLRP3 lowers the expressions of factors associated with pyroptosis in the cardiac tissue and reduces ROS production.