Cytojournal最新文献

Objective: Acute cerebral infarction (ACI) has a high incidence and complex etiology, so searching for specific diagnosis and therapeutic target molecules is highly important. This paper aimed to explore the mechanism of karyopherin alpha 4 (KPNA4) in ACI.

Material and methods: Four groups were established: sham group, middle cerebral artery occlusion (MCAO) group, MCAO + small interfering negative control (si-NC group), and MCAO + si-KPNA4 group. The Zea-Longa scoring standard was adopted to assess the neurological impairment of ACI rats. The expression of KPNA4 was verified by real-time quantity polymerase chain reaction assay and Western blot. 2,3,5-Triphenyltetrazolium chloride and hematoxylin and eosin staining were used to examine the effects of KPNA4 knockdown on brain injury in ACI rats. The apoptosis of oxygen-glucose deprivation (OGD)-SH-SY5Y cells was measured by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling method. Western blot analysis was performed to analyze the expression levels of apoptosis-related proteins, inflammatory factors, and nuclear factor kappa B (NF-κB) pathway-related proteins.

Results: KPNA4 was overexpressed in the MCAO rat brain (P < 0.0001) and OGD-SH-SY5Y cells (P < 0.0001). Knocking down KPNA4 significantly reduced the degree of brain damage (P < 0.0001) and reduced the apoptosis of OGD-SH-SY5Y cells (P < 0.001). KPNA4 knockdown also significantly decreased the expression levels of inflammatory factors (interleukin [IL]-1β, tumor necrosis factor-α, and IL-18; P < 0.0001) and prohibited the phosphorylation of the inhibitor of NF-κB and p65 (P < 0.0001). However, NF-κB agonist lipopolysaccharide reversed the inhibition of KPNA4 knockdown.

Conclusion: In general, KPNA4 knockdown can mitigate ACI-induced damage by regulating the NF-kB pathway. This finding provides a new perspective on the treatment of ACI.

Objective: Coronary microvascular dysfunction following myocardial infarction (MI) serves as a critical factor affecting cardiac repair and functional recovery. Hyperhomocysteinemia (HHcy) has been closely associated with cardiovascular diseases, particularly in terms of its detrimental effects on microvasculature post-MI. Although transcription factor SP1 plays crucial roles in various physiological and pathological processes, its specific mechanism in the reversal of HHcy-induced microvascular dysfunction after MI remains unclear. The purpose of this study was to explore the possible mechanism of SP1 on HHcy-induced microvascular dysfunction.

Material and methods: This study utilized an HHcy mouse model and an in vitro model of human coronary artery endothelial cells (HCAECs) to systematically investigate the role of SP1 in post-MI microvascular dysfunction. Cardiac microvascular perfusion was assessed using fluorescein isothiocyanate (FITC)-labeled tomato lectin. Western blot analysis was employed to examine the expression levels of signal transducer and activator of transcription 3 (STAT3), conductance calcium-activated potassium channel protein 4 (KCNN4, also known as KCa3.1), and endothelial nitric oxide synthase (eNOS). The eNOS inhibitor N(ω)-nitro-L-arginine methyl ester (L-NAME) and STAT3 inhibitor Stattic were used to validate the signaling pathway.

Results: SP1 considerably improved microvascular dysfunction and angiogenic capacity in HHcy mice after MI. It enhanced cardiac microvascular function recovery by activating the STAT3/KCa3.1/eNOS signaling pathway. The eNOS inhibitor L-NAME reversed the protective effects of SP1, which indicates the crucial role of eNOS in SP1-mediated cardiovascular protection. Furthermore, SP1 alleviated homocysteine and hypoxia-induced cytotoxicity in HCAECs through this pathway, and the inhibition of the STAT3/KCa3.1/eNOS pathway blocked SP1's protective effects.

Conclusion: This study revealed for the first time the mechanism by which SP1 reverses HHcy-induced post-MI microvascular dysfunction through the activation of the STAT3/KCa3.1/eNOS pathway. The findings not only deepen our understanding of the pathological mechanisms of post-MI microvascular dysfunction but also provide an important theoretical basis for the development of new cardiovascular disease treatment strategies. SP1, as a potential therapeutic target, may play a crucial role in future cardiovascular disease treatments.

Objective: Clear-cell renal cell carcinoma (ccRCC) is the most prevalent type of renal cancer. Solute carrier family 6 member 3 (SLC6A3) is associated with ccRCC. This research aimed to investigate the function of SLC6A3 in promoting the metabolism and development of ccRCC tumor cells.

Material and methods: SLC6A3 expression levels in ccRCC cell lines were assessed through quantitative real-time polymerase chain reaction and Western blot (WB). A stable overexpression of SLC6A3 was achieved in the 786-O cell line, and stable knockdown was established in the OS-RC-2 cell line through lentiviral transfection. Cell biological behavior was evaluated through flow cytometry, terminal deoxynucleotidyl transferase media dUTP nick end labeling staining, Cell counting kit-8 assays, and Transwell assays. Extracellular acidification rate measurements, WB, and biochemical assays were performed to detect cellular glycolysis and ferroptosis following alterations in SLC6A3 expression. The effects of SLC6A3 overexpression or knockdown on ccRCC tumor were further verified in xenograft models using nude mice.

Results: SLC6A3 was elevated in various ccRCC cell lines. The 786-O cells exhibited a relatively low baseline SLC6A3 expression (P < 0.01), which prompted subsequent overexpression experiments, whereas OSRC-2 cells showed the highest baseline expression (P < 0.001), which led to subsequent knockdown studies. The overexpression of SLC6A3 in 786-O cells increased the glycolytic activity, decreased ferroptosis levels, and promoted viability and invasion of ccRCC cells. Conversely, knockdown of SLC6A3 in OS-RC-2 cells reduced glycolytic activity, increased ferroptosis levels, and inhibited viability and invasion of ccRCC cells. All results were statistically significant. These findings were corroborated by in vivo experiments.

Conclusion: SLC6A3 is markedly overexpressed in ccRCC. It influences the promotion of tumor growth by enhancing glycolysis and suppressing ferroptosis. These insights highlight the potential of SLC6A3 for innovative therapeutic strategies in ccRCC management.

Metaplastic breast carcinoma comprises a heterogeneous group of morphologic variants characterized by markedly aggressive pathological behavior and diverse histological subtypes. Accurate diagnosis of metaplastic breast carcinoma is essential for implementing effective and individualized treatment approaches; however, its differentiation from similar tumors may occasionally be challenging. We report two cases of breast tumors diagnosed using comprehensive tumor genome analysis.

Lung cancer is the leading cause of cancer-related deaths worldwide, with genetic- and protein-based diagnostics playing a crucial role in disease detection and improving patient outcomes. Glycosylation, a major post-translational modification, has recently emerged as a factor influencing cancer progression, immune evasion, and therapeutic resistance. Aberrant glycosylation patterns, particularly among receptor tyrosine kinases (RTKs), have been shown to modulate oncogenic signaling pathways and influence tumor growth. This review provides a comprehensive overview of how glycosylation alterations affect the stability, function, and therapeutic targeting of key RTKs relevant in lung adenocarcinoma: Epidermal growth factor receptor, human epidermal growth factor receptor 2, and cellular mesenchymal-epithelial transition factor, rearranged during transfection, anaplastic lymphoma kinase, and ROS proto-oncogene 1 receptor tyrosine kinase. Despite substantial advances in targeted therapies, initial and acquired resistance remain a major challenge in the treatment of lung cancer. There is growing evidence that strategies targeting glycosylation can be combined with established treatment protocols to help overcome resistance. Finally, we propose future directions for the advancement of glycosylation-based approaches to improve precision medicine.

Objective: This study aims to explore the role of microfibrillar-associated protein-2 (MFAP2) in oral squamous cell carcinoma (OSCC).

Material and methods: Analysis of MFAP2 expression in diverse cancers and its relationship with head-and-neck squamous cell carcinoma (HNSC) prognosis. MFAP2 abundance was identified in OSCC cells and in human oral epithelial cells using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot assays. Knockdown and overexpression techniques were utilized to examine the mechanism by which MFAP2 and nuclear factor erythroid 2-related factor 2 (NRF2) affect OSCC malignancy. Cell viability, proliferation, and apoptosis were assessed using cell counting kit-8, colony formation, flow cytometry, wound healing, and Transwell tests. Messenger ribonucleic acid expression was detected using qRT-PCR, whereas protein level was analyzed using Western blot.

Results: MFAP2 and Kelch-like E3 ubiquitin ligase (ECH)-associated protein 1 (KEAP1) had high expression levels in numerous tumors, including OSCC, and the high expression level of MFAP2 was associated with unfavorable HNSC outcomes. MFAP2 was abundantly expressed in five OSCC cell lines, with the peak expression observed in squamous cell carcinoma (SCC)-15 and SCC-9 cells, making them suitable for subsequent studies. MFAP2 knockdown hindered the proliferative and mobile capacity of OSCC cells, yet it supported cell apoptosis. MFAP2 silencing led to a notable drop in KEAP1 and NRF2 expression levels in OSCC cells. NRF2 overexpression could counteract the effects of MFAP2 knockout, which included diminished proliferation and movement and heightened apoptosis in OSCC cells.

Conclusion: The results of this study indicated that MFAP2 facilitated the malignant progression of OSCC and provided insights into the downstream regulatory mechanism of the KEAP1/NRF2 axis, highlighting the potential application of MFAP2 in OSCC management.

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