Background The poor prognosis of ovarian cancer patients is strongly related to peritoneal metastasis with the production of malignant ascites. However, it remains largely unclear how ascites in the peritoneal cavity influences tumor metabolism and recurrence. This study is an explorative approach aimed at for a deeper molecular and physical–chemical characterization of malignant ascites and to investigate their effect on in vitro ovarian cancer cell proliferation. Methods This study included 10 malignant ascites specimens from patients undergoing ovarian cancer resection. Ascites samples were deeply phenotyped by 1H-NMR based metabolomics, blood-gas analyzer based gas flow analysis and flow cytomertry based a 13-plex cytokine panel. Characteristics of tumor cells were investigated in a 3D spheroid model by SEM and metabolic activity, adhesion, anti-apoptosis, migratory ability evaluated by MTT assay, adhesion assay, flowcytometry and scratch assay. The effect of different pH values was assessed by adding 10% malignant ascites to the test samples. Results  The overall extracellular (peritoneal) environment was alkaline, with pH of ascites at stage II-III = 7.51 ± 0.16, and stage IV = 7.78 ± 0.16. Ovarian cancer spheroids grew rapidly in a slightly alkaline environment. Decreasing pH of the cell culture medium suppressed tumor features, metabolic activity, adhesion, anti-apoptosis, and migratory ability. However, 10% ascites could prevent tumor cells from being affected by acidic pH. Metabolomics analysis identified stage IV patients had significantly higher concentrations of alanine, isoleucine, phenylalanine, and glutamine than stage II-III patients, while stage II-III patients had significantly higher concentrations of 3-hydroxybutyrate. pH was positively correlated with acetate, and acetate positively correlated with lipid compounds. IL-8 was positively correlated with lipid metabolites and acetate. Glutathione and carnitine were negatively correlated with cytokines IL-6 and chemokines (IL-8 & MCP-1). Conclusion Alkaline malignant ascites facilitated ovarian cancer progression. Additionally, deep ascites phenotyping by metabolomics and cytokine investigations allows for a refined stratification of ovarian cancer patients. These findings contribute to the understanding of ascites pathology in ovarian cancer.

Background: Genes of the centromere protein (CENP) are involved in the kinetochore assembly during mitosis. The expression of some CENPgenes has been associated with various carcinogenesis and development. However, it is unclear whether abnormal expression of CENP genes as a unit affects the clinical outcome, immune cell infiltration, and immunotherapy of breast cancer (BRCA) patients. Methods: We systematically evaluated CENP genes and comprehensively determined the correlation between high and low expression and prognosis, staging, malignant phenotype, and drug sensitivity of BRCA. The correlation between CENP genes and immune cell infiltration was comprehensively determined. Further screening for immune- and metabolic-related hub genes was used to quantify subtypes of patients. Then, we evaluated their value in prognosis prediction and treatment response to BRCA. The correlation between CENP genes and drug resistance was also determined by sequencing the tissues of patients. Single-cell sequencing to further analyze CENP genes expression in metastatic cancer. Results: The CENP genes were significantly differentially expressed in BRCA compared to para-cancer. Highly expressed CENP genes had a poor prognosis, late stage, and a high proportion of malignant phenotypes. We identified high and low CENPclusters and observed that high expression was associated with poor prognosis, high immune escape, and reduced drug sensitivity in patients. Afterward, we screened immune- and metabolic-related hub genes and predicted the prognosis of BRCA patients based on them. Furthermore, CENP genes were elevated in drug-resistant breast cancer tissues and metastatic breast cancer tissues. Conclusion: Collectively, we determined the prognostic impact of the CENP genes on BRCA patients. Furthermore, by further screening the hub genes to subgroup breast cancer patients to predict prognosis and to be able to explore more effective treatment strategies based on the expression of CENP genes.

Background The poor prognosis of ovarian cancer patients is strongly related to peritoneal metastasis with the production of malignant ascites. However, it remains largely unclear how ascites in the peritoneal cavity influences tumor metabolism and recurrence. This study is an explorative approach aimed at for a deeper molecular and physical-chemical characterization of malignant ascites and to investigate their effect on in vitro ovarian cancer cell proliferation. Methods This study included 10 malignant ascites specimens from patients undergoing ovarian cancer resection. Ascites samples were deeply phenotyped by 1H-NMR based metabolomics, gas flow analysis and a 13-plex cytokine panel. Characteristics of tumor cells were investigated in a 3D spheroid model by metabolic activity, adhesion, anti-apoptosis, migratory ability adhesion assay, flowcytometry and scratch assay. The effect of different pH values was assessed by adding 10% malignant ascites to the test samples. Results The overall extracellular (peritoneal) environment was alkaline, with pH of ascites at stage II-III = 7.51 ± 0.16, and stage IV = 7.78 ± 0.16. Ovarian cancer spheroids grew rapidly in a slightly alkaline environment. Decreasing pH of the cell culture medium suppressed tumor features, metabolic activity, adhesion, anti-apoptosis, and migratory ability. However, 10% ascites could prevent tumor cells from being affected by acidic pH. Metabolomics analysis identified stage IV patients had significantly higher concentrations of alanine, isoleucine, phenylalanine, and glutamine than stage II-III patients, while stage II-III patients had significantly higher concentrations of 3-hydroxybutyrate. Various positive and negative correlations were observed between physical-chemical parameters, polar/lipid metabolites, and cytokines. Conclusion Malignant ascites facilitated ovarian cancer progression. Additionally, deep ascites phenotyping by metabolomics and cytokine investigations allows for a refined stratification of ovarian cancer patients.