Unveiling the methionine cycle: a key metabolic signature and NR4A2 as a methionine-responsive oncogene in esophageal squamous cell carcinoma.

Esophageal squamous cell carcinoma (ESCC) is a deadly malignancy with notable metabolic reprogramming, yet the pivotal metabolic feature driving ESCC progression remains elusive. Here, we show that methionine cycle exhibits robust activation in ESCC and is reversely associated with patient survival. ESCC cells readily harness exogenous methionine to generate S-adenosyl-methionine (SAM), thus promoting cell proliferation. Mechanistically, methionine augments METTL3-mediated RNA m6A methylation through SAM and revises gene expression. Integrative omics analysis highlights the potent influence of methionine/SAM on NR4A2 expression in a tumor-specific manner, mediated by the IGF2BP2-dependent stabilization of methylated NR4A2 mRNA. We demonstrate that NR4A2 facilitates ESCC growth and negatively impacts patient survival. We further identify celecoxib as an effective inhibitor of NR4A2, offering promise as a new anti-ESCC agent. In summary, our findings underscore the active methionine cycle as a critical metabolic characteristic in ESCC, and pinpoint NR4A2 as a novel methionine-responsive oncogene, thereby presenting a compelling target potentially superior to methionine restriction.

Active post-transcriptional regulation and ACLY-mediated acetyl-CoA synthesis as a pivotal target of Shuang-Huang-Sheng-Bai formula for lung adenocarcinoma treatment.

BACKGROUND: New therapeutic approaches are required to improve the outcomes of lung cancer (LC), a leading cause of cancer-related deaths worldwide. Chinese herbal medicine formulae widely used in China provide a unique opportunity for improving LC treatment, and the Shuang-Huang-Sheng-Bai (SHSB) formula is a typical example. However, the underlying mechanisms of action remains unclear. PURPOSE: This study aimed to confirm the efficacy of SHSB against lung adenocarcinoma (LUAD), which is a major histological type of LC, unveil the downstream targets of this formula, and assess the clinical relevance and biological roles of the newly identified target. METHODS: An experimental metastasis mouse model and a subcutaneous xenograft mouse model were used to evaluate the anti-cancer activity of SHSB. Multi-omics profiling of subcutaneous tumors and metabolomic profiling of sera were performed to identify downstream targets, especially the metabolic targets of SHSB. A clinical trial was conducted to verify the newly identified metabolic targets in patients. Next, the metabolites and enzymes engaged in the metabolic pathway targeted by SHSB were measured in clinical samples. Finally, routine molecular experiments were performed to decipher the biological functions of the metabolic pathways targeted by SHSB. RESULTS: Oral SHSB administration showed overt anti-LUAD efficacy as revealed by the extended overall survival of the metastasis model and impaired growth of implanted tumors in the subcutaneous xenograft model. Mechanistically, SHSB administration altered protein expression in the post-transcriptional layer and modified the metabolome of LUAD xenografts. Integrative analysis demonstrated that SHSB markedly inhibited acetyl-CoA synthesis in tumors by post-transcriptionally downregulating ATP-citrate lyase (ACLY). Consistently, our clinical trial showed that oral SHSB administration declined serum acetyl-CoA levels of patients with LC. Moreover, acetyl-CoA synthesis and ACLY expression were both augmented in clinical LUAD tissues of patients, and high intratumoral ACLY expression predicted a detrimental prognosis. Finally, we showed that ACLY-mediated acetyl-CoA synthesis is essential for LUAD cell growth by promoting G1/S transition and DNA replication. CONCLUSION: Limited downstream targets of SHSB for LC treatment have been reported in previous hypothesis-driven studies. In this study, we conducted a comprehensive multi-omics investigation and demonstrated that SHSB exerted its anti-LUAD efficacy by actively and post-transcriptionally modulating protein expression and particularly restraining ACLY-mediated acetyl-CoA synthesis.

Promoter methylation of glutathione S-transferase pi1 and multidrug resistance gene 1 in bronchioloalveolar carcinoma and its correlation with DNA methyltransferase 1 expression.

BACKGROUND: The presence of glutathione S-transferase (GST) pi1 (GSTP1) or multidrug resistance gene 1 (MDR1) promoter methylation in lung cancer was studied for the first time to the authors' knowledge; and, to date, the clinical significance of methylation is not clear. The objective of the current study was to determine the promoter methylation status of GSTP1 and MDR1, which encode GST-pi and P-glycoprotein (Pgp), respectively, in patients with bronchioloalveolar carcinoma (BAC) and to investigate whether methyltransferase 1 (DNMT1)-mediated GSTP1 or MDR1 methylation are responsible for disease progression and prognosis in patients with BAC. METHODS: Protein expression levels of DNTM1, GST-pi, and Pgp were determined by immunohistochemistry in samples from 36 patients with BAC. Promoter methylation status of the GSTP1 and MDR1 genes was determined by using methylation-specific polymerase chain reaction analysis. RESULTS: The results demonstrated a significant correlation between the methylation of the GSTP1 or MDR1 promoters and negative expression of their respective proteins in BAC (P < .05). A significant correlation also was demonstrated between GSTP1 methylation and recurrence-free and overall survival of patients with BAC. DNMT1 protein expression levels were correlated with GSTP1 promoter methylation and patient prognosis (P < .05). However, no correlation was observed between DNMT1 expression and MDR1 methylation. CONCLUSIONS: GSTP1 promoter methylation mediated by DNMT1 may promote BAC progression and could serve as a poor prognostic indicator for patients with this disease. DNMT1 protein expression also may be considered as a prognostic indicator. Methylation of the MDR1 promoter may be mediated through pathways other than DNMT1 in BAC and does not appear to be associated with disease progression or patient prognosis.

[Microvessel density and expressions of survivin and vascular endothelial growth factor in non-small cell lung cancer and their correlations to clinicopathologic features].

BACKGROUND & OBJECTIVE: Survivin, an anti-apoptosis gene, expresses in most tumors, and takes part in tumor angiogenesis. This study was to investigate microvessel density (MVD) and expressions of Survivin and vascular endothelial growth factor (VEGF) in non-small cell lung cancer (NSCLC), and explore their correlations to clinicopathologic features of NSCLC. METHODS: MVD and expressions of Survivin and VEGF in 96 specimens of NSCLC tissues, 31 specimens of tumor adjacent tissues, and 20 specimens of benign lesions were detected by SP immunohistochemistry; their interrelations and correlations to clinicopathologic features of NSCLC were analyzed. RESULTS: Positive rate of Survivin was significantly higher in NSCLC than in adjacent tissues and benign lesions (69.8% vs. 16.1% and 0, P<0.05); its expression was related with differentiation and TNM stage of NSCLC. Positive rate of VEGF was significantly higher in NSCLC than in adjacent tissues and benign lesions (72.9% vs. 45.2% and 25.0%, P<0.05); its expression was related with lymph node metastasis and TNM stage of NSCLC. MVD was significantly higher in NSCLC than in adjacent tissues and benign lesions (24.44+/-7.79 vs. 19.37+/-5.26 and 11.83+/-6.25, P<0.05), and was related with lymph node metastasis and TNM stage of NSCLC. Survivin expression was positively correlated with VEGF expression and MVD. CONCLUSION: Survivin is overexpressed in NSCLC, which relates with differentiation and TNM stage of NSCLC and takes part in angiogenesis.

[Changes of serum interleukin (IL)-12 and IL-13 in asthmatic patients and regulatory effects of glucocorticoids on them].

OBJECTIVE: To observe the changes of serum interleukin (IL)-12, IL-13 and lung function in asthmatic patients and to evaluate the influence of glucocorticoid on them. METHODS: The serum samples were obtained from (1) 25 asthmatic patients with acute asthma attack before and after one week course of oral prednisone, (2) 20 asthmatic patients in remission stage, and (3) 15 healthy volunteers. Serum IL-12 and IL-13 were determined with sandwich ELISA. Lung ventilatory function forced expiratory volume in one second (FEV(1)) and respiratory impedance airway resistance (R(5)) were measured in all patients. RESULTS: (1) Serum level of IL-12 in asthma attack group was significantly lower than that in asthma remission group (P < 0.05); both were significantly lower than that in normal control group (P < 0.01); in asthma attack group IL-12 level after prednisone treatment was significantly higher than that before treatment (P < 0.01). (2) Serum level of IL-13 in asthma attack group was significantly higher than that in asthma remission group (P < 0.01); both were significantly higher than that in normal control group (P < 0.01); in asthma attack group IL-13 level after prednisone treatment was significantly lower than that before treatment (P < 0.01). Correlation analysis showed that the serum level of IL-12 was positively correlated to FEV(1) and negatively correlated to R(5) and to serum level of IL-13 (r(1) = 0.458, r(2) = -0.516, and r(3) = -0.549, respectively; P < 0.05 and P < 0.01); the serum level of IL-13 was negatively correlated to FEV(1) and positively correlated to R(5) (r(1) = -0.493, and r(2) = 0.528, respectively; P < 0.05). CONCLUSION: The secretion of IL-12 and IL-13 was impaired in asthma with a significant increase in serum level of IL-13 and decrease in serum level of IL-12. Glucocorticoid could downregulate the serum level of IL-13 and upregulate the serum level of IL-12, redress the imbalance of IL-12/IL-13, and improve lung function in asthmatic patients.