Translational cancer research最新文献

Background: Chemotherapy plays a crucial role in the treatment of advanced hepatocellular carcinoma (HCC), but the therapeutic efficacy of cisplatin, a commonly used chemotherapeutic agent, is frequently compromised by drug resistance in advanced stages of the disease. A previous study demonstrated that the combination of emodin and cisplatin alleviates cisplatin resistance by inhibiting epithelial-mesenchymal transition (EMT), but the underlying molecular mechanism remains unclear. This study aimed to systematically elucidate the mechanism of action of emodin in enhancing the antitumor activity of cisplatin.

Methods: The synergistic concentration of emodin and cisplatin was determined using the CCK-8 assay, combined with transcriptome sequencing to analyze the differentially expressed genes and signaling pathways, and Western blot (WB) to validate the expression of key proteins.

Results: The combination of emodin (50 µM) and cisplatin (10 µM) inhibited the proliferation, invasion, and migration of HepG2 cells. Transcriptomic analysis revealed that the combination exerted a synergistic effect through the regulation of the Rap1 pathway. Pathway inhibition assays verified that the combination downregulated Rap1, vimentin, N-cadherin, and p-AKT/AKT, while upregulating the expression of the epithelial marker E-cadherin.

Conclusions: Emodin potentiates the anti-tumor efficacy of cisplatin against HCC while suppressing metastasis, mechanistically through targeted inhibition of the Rap1 signaling pathway and subsequent blockade of the EMT.

Background: Lactate and lactylation-related genes (LRGs) have demonstrated promising effects in mitigating tumor growth and improving clinical outcomes, but their role in the ovarian cancer (OV) microenvironment and immunotherapy has not been extensively examined. This study aimed to characterize the relationship between LRGs and the outcomes of patients with OV.

Methods: We collected bulk RNA-sequencing data from The Cancer Genome Atlas (TCGA) OV dataset via the University of California, Santa Cruz (UCSC) Xena and bulk RNA fragments per kilobase of transcript per million mapped reads (FPKM) sequences of normal ovarian tissues via the Genotype-Tissue Expression (GTEx) project. We then analyzed the relationship of 22 LRGs with programmed cell death protein 1 (PD-1) or programmed death-ligand 1 (PD-L1) and compared the prognosis of patients with different expressions of LRGs in response to anti-PD-1/PD-L1 immunotherapy. We used the hierarchical clustering method to delineate subtypes of patients with OV according to LRG and PD-1 and PD-L1 expression and compared their tumor microenvironment (TME) compositions. A neural network was trained based on LRGs to predict the immunotherapy response in patients with OV, and single-cell RNA (scRNA) analysis was used to clarify the mechanisms underlying the LRGs' predictive capacity. Meanwhile, a tissue array was used to determine the LRG hub genes' influence on the prognosis of patients with OV.

Results: Most of the LRGs examined were associated with the prognosis of patients with OV undergoing anti-PD-1/PD-L1 immunotherapy. According to the expression of the LRG panel, patients with OV could be clustered in subtypes, with each cluster exhibiting a distinct TME composition and immune-cell ratio. The neural network based on LRG expression could predict the immunotherapy response of patients with OV, with LDHA and LDHB being the potential hub LRGs. Patients with OV and low LDHA expression had better prognosis in terms of disease-free survival (DFS) or overall survival (OS), while patients with OV and high LDHB expression had a longer DFS and OS.

Conclusions: The expression of lactylation-related hub genes can be used to stratify patients with OV and predict their response to immunotherapy.

Background: Nasopharyngeal carcinoma (NPC) is a widely prevalent malignant tumor with a marked tendency toward metastasis and recurrence. Ferroptosis-related genes (FRGs) are critically involved in the pathogenesis of NPC. This study aims to employ bioinformatics analysis methods to identify key genes influencing the malignant progression of NPC and to investigate the regulatory mechanisms of these genes.

Methods: Bulk RNA sequencing datasets (GSE53819, GSE61218, GSE64634, GSE12452, and GSE102349) and a single-cell RNA sequencing dataset (GSE150825) were downloaded from the Gene Expression Omnibus. Integrated bioinformatics analyses-including differential expression analysis, weighted gene co-expression network analysis, machine learning, and survival analysis-were conducted to identify key FRGs associated with NPC. Intracellular expression levels of adenosylhomocysteinase (AHCY), acyl-CoA synthetase long chain family member 4, glutathione peroxidase 4, macrophage stimulating 1, and Yes1 associated transcriptional regulator were detected through western blot. Cell viability was assessed using the cell counting kit-8; cell death was determined by flow cytometry; and cell migration and invasion were evaluated using wound-healing and Transwell assays. Intracellular reactive oxygen species levels were determined using the fluorescent probe 2',7'-dichlorodihydrofluorescein diacetate, and malondialdehyde and glutathione levels were detected using their respective detection kits.

Results: Four key FRGs-isocitrate dehydrogenase 1, AHCY, endothelial PAS domain-containing protein 1 (EPAS1), and ARHGEF26 antisense RNA 1-were identified. Survival analysis of publicly available cohorts highlighted AHCY and EPAS1 as potential biomarkers for survival in patients with NPC. We selected AHCY for further in vitro mechanistic analysis and found it to be upregulated in NPC cell lines (NPC/HK1 and c666-1) relative to the nasopharyngeal epithelial cell line NP69. Functionally, AHCY knockdown in c666-1 cells inhibited cell migration, viability, and invasion, as well as suppressing the Hippo-Yes-associated protein (YAP) pathway, while promoting ferroptosis. Conversely, AHCY overexpression in NPC/HK1 cells enhanced cell migration, viability, and invasion, as well as promoting the Hippo-YAP pathway, while inhibiting ferroptosis. Treatment of AHCY-knockdown c666-1 cells with the ferroptosis inhibitor ferrostatin-1 enhanced cell viability, migration, and invasion, while treatment with the Hippo-YAP pathway agonist PY-60 promoted cell viability, migration, and invasion, while inhibiting ferroptosis.

Conclusions: Our in vitro findings indicate that AHCY suppresses ferroptosis, partly via the Hippo-YAP pathway, thereby promoting the invasion and migration of NPC cells. Further in vivo and clinical studies are warranted to validate these findings.

Background: Hepatocellular carcinoma (HCC) is a major cause of cancer-related mortality. Surgical resection offers a curative option, but outcomes vary widely. Accurate preoperative risk stratification is essential. This study evaluated the prognostic value of the albumin-bilirubin (ALBI) score and triglyceride-glucose (TyG) index-markers of liver function and metabolic status-in patients undergoing curative liver resection.

Methods: This retrospective study included 238 patients with pathologically confirmed HCC who underwent curative liver resection. Preoperative clinical and biochemical variables were collected, and a prognostic model was developed using the entire dataset. Multivariate Cox proportional hazards regression was performed to identify independent prognostic factors. A nomogram incorporating the ALBI score and TyG index was constructed to estimate 2-, 3-, and 4-year overall survival (OS). Model performance was assessed through Kaplan-Meier survival analysis, receiver operating characteristic (ROC) curves, calibration plot, and decision curve analysis (DCA). Bootstrap resampling was used for internal validation to generate corrected estimates of model accuracy.

Results: Multivariate Cox regression demonstrated that both the ALBI score [hazard ratio (HR) =5.120; 95% confidence interval (CI): 2.944-8.905; P<0.001] and the TyG index (HR =4.202; 95% CI: 2.459-7.180; P<0.001) were independent predictors of OS. Patients with elevated ALBI or TyG values exhibited markedly shorter median survival compared with those with lower values. The ALBI-TyG nomogram showed robust discriminative ability, yielding an area under the ROC curve of 0.823 (95% CI: 0.768-0.878), outperforming either marker alone as well as traditional liver function scoring systems. Calibration plot demonstrated good agreement between predicted and observed survival probabilities, while DCA indicated superior net clinical benefit across a wide range of threshold probabilities.

Conclusions: Combining ALBI score and TyG index offers an objective and accessible tool for prognostic assessment in HCC patients undergoing liver resection, enabling more personalized perioperative management.

Background: Cervical cancer remains a major malignancy seriously threatening women's health. Rosa roxburghii Tratt fruit polysaccharides (RTFP), a natural extract possessing diverse biological activities, can inhibit tumors through mechanisms such as inducing apoptosis and cell cycle arrest. However, the potential effects of RTFP on cervical cancer and the associated molecular mechanisms remain incompletely elucidated. Therefore, this study aimed to investigate the effects of RTFP sourced on the proliferation, migration, invasion, and apoptosis of human cervical cancer cell line.

Methods: Cell viability was assessed using the Cell Counting Kit-8 (CCK-8) assay. HeLa cells were treated with RTFP at concentrations of 2, 4, 6, 8, and 10 mg/mL for 24, 48, and 72 h. Cell migration and invasion were evaluated using scratch wound healing and Transwell assays, respectively, after exposure to RTFP (3 and 6 mg/mL) for 24 and 48 h. For cell cycle and apoptosis analysis, cells treated with RTFP (3 and 6 mg/mL) for 48 h were analyzed by flow cytometry. The messenger RNA (mRNA) and protein expression levels of cell cycle regulators (CDK-1 and cyclin B1) and apoptosis-related markers (Bax, Bcl-2, caspase-3, caspase-8, and caspase-9) were quantified by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively, following 48 h of treatment with RTFP (3 and 6 mg/mL). All experiments were performed in three independent replicates (n=3), and data are presented as mean ± standard deviation (SD).

Results: RTFP inhibited the proliferation, migration, and invasion of HeLa cells in a time- and dose-dependent manner (P<0.01). It induced cell cycle arrest at the G2/M phase (P<0.01) and downregulated the protein expression of CDK-1 and cyclin B1 (P<0.05). Additionally, RTFP promoted apoptosis through modulating apoptosis-related proteins: it upregulated the pro-apoptotic proteins Bax, caspase-3, caspase-8, and caspase-9 (P<0.05), while downregulated the anti-apoptotic protein Bcl-2 (P<0.01).

Conclusions: RTFP inhibits the proliferation, migration, and invasion of HeLa cells, arrests the cell cycle at the G2/M phase, and induces apoptosis. These in vitro findings preliminarily demonstrate the potential anti-cervical cancer activity of RTFP in HeLa cells, warranting further in-depth investigations.

Background: Angiogenesis plays a pivotal role in driving tumour progression, including tumour growth, local invasion, and distant metastasis. Calponin 3 (CNN3) belongs to the actin-binding protein family and has been shown to be aberrantly expressed in osteosarcoma specimens, but its exact role in the regulation of angiogenesis remains unknown. This study aimed to investigate the function and underlying mechanism of CNN3 in osteosarcoma angiogenesis.

Methods: CNN3 and CD31 expression in osteosarcoma specimens was detected by immunohistochemistry. CNN3 in MG-63 and Saos-2 cells was silenced or overexpressed by transfection with small interfering RNA or overexpression plasmids, respectively. Vascular endothelial growth factor-A (VEGF-A) levels were determined by enzyme-linked immunosorbent assay. Human umbilical vein endothelial cells (HUVECs) were co-cultured with CNN3-silenced or -overexpressing osteosarcoma cells using a Transwell chamber. The function of HUVECs were assessed using cell counting, scratch wound healing, Transwell Matrigel invasion, and Matrigel vascular mimicry assays. The protein levels of endothelial-to-mesenchymal transition (EndMT) markers and phosphorylated Akt in HUVECs was determined using western blotting.

Results: Immunohistochemical analysis demonstrated a significant positive correlation between CNN3 expression and CD31 (microvascular density marker) levels in osteosarcoma tissues (r=0.7264, P=0.0003). HUVECs co-cultured with CNN3-overexpressing cells exhibited enhanced proliferative, migratory, invasive, and tube-forming capacities along with promoted EndMT, whereas CNN3 silencing suppressed these effects. CNN3 silencing reduced, while its overexpression elevated, VEGF-A secretion in both MG-63 and Saos-2 cells. Furthermore, silencing CNN3 in MG-63 and Saos-2 cells decreased Akt phosphorylation in co-cultured HUVECs, whereas CNN3 overexpression concomitantly increased it.

Conclusions: CNN3 promotes osteosarcoma angiogenesis, possibly through upregulating VEGF-A secretion and enhancing endothelial cell functions, suggesting its potential as the therapeutic target for anti-angiogenic strategies.