Previous studies showed that sodium ferulate, the effective component of Chinese herb, can inhibit platelet aggregation and decrease serum lipid. However, it is still unknown if sodium ferulate could prevent atherosclerogenesis. The experiments were designed to study its effects and mechanisms on atherosclerogenesis. Blood samples and thoracic aortas obtained from Japanese rabbits fed by high-lipid or high-lipid plus sodium ferulate forage were analyzed and compared. Simultaneously, vascular endothelial cells were cultured and treated by hyperlipidemic serum solely or plus sodium ferulate. Cellular ultrastructure, nitric oxide (NO) production, and cytokines expressions were studied and compared. In vivo experiment, aorta atherosclerotic plaque area of sodium ferulate-treated rabbits was much smaller than that of high-lipid-fed rabbits and serum triglyceride was correlated positively with the plaque area in both groups. In vitro, endothelial cells incubated with hyperlipidemic serum exhibited pronounced ultrastructural abnormalities, transforming growth factor beta1 expression and NO release were significantly decreased, while basic fibroblast growth factor expression was increased. Interestingly, the treatment group results clearly demonstrated that sodium ferulate was effective to protect cells from detrimental effects of hyperlipidemic serum and to help maintain normal NO and cytokines expressions. We concluded that sodium ferulate could inhibit rabbit aorta atherosclerogenesis, possibly through decreasing the serum lipid concentration and preventing vascular endothelial cells from the injury of hyperlipidemic serum.

Background Lung cancer, one of the most common malignant tumors, exhibits high inter- and intra-tumor heterogeneity which contributes significantly to treatment resistance and failure. Single-cell RNA sequencing (scRNA-seq) has been widely used to dissect the cellular composition and characterize the molecular properties of cancer cells and their tumor microenvironment in lung cancer. However, the transcriptomic heterogeneity among various cancer cells in non-small cell lung cancer (NSCLC) warrants further illustration. Methods To comprehensively analyze the molecular heterogeneity of NSCLC, we performed high-precision single-cell RNA-seq analyses on 7364 individual cells from tumor tissues and matched normal tissues from 19 primary lung cancer patients and 1 pulmonary chondroid hamartoma patient. Results In 6 of 16 patients sequenced, we identified a significant proportion of cancer cells simultaneously expressing classical marker genes for two or even three histologic subtypes of NSCLC—adenocarcinoma (ADC), squamous cell carcinoma (SCC), and neuroendocrine tumor (NET) in the same individual cell, which we defined as mixed-lineage tumor cells; this was verified by both co-immunostaining and RNA in situ hybridization. These data suggest that mixed-lineage tumor cells are highly plastic with mixed features of different types of NSCLC. Both copy number variation (CNV) patterns and mitochondrial mutations clearly showed that the mixed-lineage and single-lineage tumor cells from the same patient had common tumor ancestors rather than different origins. Moreover, we revealed that patients with high mixed-lineage features of different cancer subtypes had worse survival than patients with low mixed-lineage features, indicating that mixed-lineage tumor features were associated with poorer prognosis. In addition, gene signatures specific to mixed-lineage tumor cells were identified, including AKR1B1. Gene knockdown and small molecule inhibition of AKR1B1 can significantly decrease cell proliferation and promote cell apoptosis, suggesting that AKR1B1 plays an important role in tumorigenesis and can serve as a candidate target for tumor therapy of NSCLC patients with mixed-lineage tumor features. Conclusions In summary, our work provides novel insights into the tumor heterogeneity of NSCLC in terms of the identification of prevalent mixed-lineage subpopulations of cancer cells with combined signatures of SCC, ADC, and NET and offers clues for potential treatment strategies in these patients.

The HeLa cells are the earliest and mostly used laboratory human cells for biomedical particularly cancer research. They were derived from a patient’s cervical cancerous tissue, and are known for their heterogeneous cellular origin and variable genomic landscapes. Single-cell sequencing techniques with faithful linear and uniformly amplified genomes (DNA) and transcriptomes (RNA) may facilitate the study of cellular differences at the individual cell level. In this work, we have performed single-cell DNA and RNA sequencing with HeLa-CCL2 cells to study their heterogeneity. We have studied the complexity of copy number variations (CNVs) of HeLa-CCL2 genome at the single cell level, and revealed the transcriptomic heterogeneity of HeLa-CCL2. We also analyzed the relationship between genome and transcriptome at the single-cell level, and found overall correlation between CNV and transcriptome expression patterns. Finally, we concluded that although single-cell sequencing techniques are applicable to study heterogeneous cells such as HeLa-CCL2, the data analyses need to be more careful and well controlled.