Anti-cancer activity and mechanism of flurbiprofen organoselenium compound RY-1-92 in non-small cell lung cancer.

Lung cancer is one of the malignancies with the highest incidence and mortality rates worldwide, and non-small cell lung cancer (NSCLC) accounts for about 85% of all lung cancer types. In this study, the anti-cancer activities of a novel flurbiprofen organic selenium compound, RY-1-92, on NSCLC cells and a mouse model and the underlying molecular mechanisms were explored. We found that compound RY-1-92 can significantly inhibit the viability, colony formation and migration of A549, NCI-H460 lung cancer cells. Flow cytometry analysis showed that RY-1-92 also can lead to G2/M cell cycle arrest and apoptosis induced in lung cancer cells. Further, RY-1-92 can decrease the tumor size in the Lewis lung cancer tumor-bearing mouse model. The protein levels of cell cycle-related proteins CDK1/cyclinB1 were decreased, while the apoptosis-related protein BAX was increased dramatically after RY-1-92 treatment in vitro and in vivo. Impressively, it was found that TRPV1 might act as a potential molecular target of RY-1-92 using the SEA search server. Furthermore, down-regulation on TRPV1 and its downstream associated factors including p-AKT protein and MAPK signaling pathway-related proteins after RY-1-92 treatment was observed in A549, NCI-H460 lung cancer cells. Taken together, our findings shed light on the potential of RY-1-92 as a novel small molecular drug for NSCLC, and it is of great significance for its further in-depth research and development.

Comparative Analysis of Key Transcription Factors Involved in Development in Sea Lamprey (Petromyzon marinus) through Systematic Mining of Lamprey Gene Expression Dataset.

Transcription factors (TFs), key regulators for gene expression, play varied yet crucial roles throughout cellular polarization, migration, proliferation, differentiation and apoptosis. An important function of TFs is acting in embryogenesis and organogenesis. To identify the candidate TFs in the progression of lamprey early embryogenesis, datasetGSE76037 was downloaded from the Gene Expression Omnibus (GEO) database and an integrated bioinformatic analysis was performed. Our findings revealed a total of 152 TFs in the dataset. The function enrichment analysis showed that these genes were mainly enriched in transcription from RNA polymerase II promoter, cell differentiation, embryonic digestive tract morphogenesis and so on. Hierarchical clustering analysis suggested the expression of TFs was significantly different during early embryogenesis. Moreover, volcano plots and Venn diagrams analysis identified 36 key TFs, which were considered to play an important role during embryogenesis. The weighted correlation network analysis (WGCNA) was constructed and the Pearson correlation coefficient was performed, indicating these TFs might involve in early development of lamprey germline by synergistically regulating each other. The result was confirmed by real-time polymerase chain reaction and Western blotting analysis. In conclusion, differentially expressed genes identified in the present study help us understand the molecular mechanisms underlying the lamprey embryogenesis and provide candidate TFs for further study of vertebrate embryonic development.

Molecular Evolution of Transforming Growth Factor-ß (TGF-ß) Gene Family and the Functional Characterization of Lamprey TGF-ß2.

The transforming growth factor-ßs (TGF-ßs) are multifunctional cytokines capable of regulating a wide range of cellular behaviors and play a key role in maintaining the homeostasis of the immune system. The TGF-ß subfamily, which is only present in deuterostomes, expands from a single gene in invertebrates to multiple members in jawed vertebrates. However, the evolutionary processes of the TGF-ß subfamily in vertebrates still lack sufficient elucidation. In this study, the TGF-ß homologs are identified at the genome-wide level in the reissner lamprey (Lethenteron reissneri), the sea lamprey (Petromyzon marinus), and the Japanese lamprey (Lampetra japonica), which are the extant representatives of jawless vertebrates with a history of more than 350 million years. The molecular evolutionary analyses reveal that the lamprey TGF-ß subfamily contains two members representing ancestors of TGF-ß2 and 3 in vertebrates, respectively, but TGF-ß1 is absent. The transcriptional expression patterns show that the lamprey TGF-ß2 may play a central regulatory role in the innate immune response of the lamprey since it exhibits a more rapid and significant upregulation of expression than TGF-ß3 during lipopolysaccharide stimuli. The incorporation of BrdU assay reveals that the lamprey TGF-ß2 recombinant protein exerts the bipolar regulation on the proliferation of the supraneural myeloid body cells (SMB cells) in the quiescent and LPS-activated state, while plays an inhibitory role in the proliferation of quiescent and activated leukocytes in lampreys. Furthermore, caspase-3/7 activity analysis indicates that the lamprey TGF-ß2 protects SMB cells from apoptosis after serum deprivation, in contrast to promoting apoptosis of leukocytes. Our composite results offer valuable clues to the origin and evolution of the TGF-ß subfamily and imply that TGF-ßs are among the most ancestral immune regulators in vertebrates.

Adaptive immune stimuli altered the cargo proteins of exosomes released by supraneural myeloid body cells in Lampetra japonica.

As one of the representatives of jawless vertebrates, lamprey is an important animal model for research on the evolution of adaptive immune system. Although it is widely accepted that the key characteristic of the immune response in jawed vertebrates is the functional cooperation between different immune effector cell lineages, whether immune cells of lamprey can communicate with each other is still unclear. Recently, mounting evidence has revealed the emerging role of exosomes in mediating intercellular communication. In this study, by means of ultrafiltration followed by size exclusion chromatography, exosomes are purified from conditioned growth medium of the primary supraneural myeloid body cells (SMB cells) in LPS-immunized and control Lampetra japonica (Japanese lamprey). The proteomic profiling and qualitative comparison are performed between protein components enriched in exosomes released by SMB cells under quiescent and activated conditions. Our results demonstrate that SMB cells can release exosomes with well-defined characteristics comparable to their mammalian counterparts on their size, morphology and protein markers, which supports exosomes are evolutionarily conserved between lamprey and other higher vertebrate species. In terms of comparison with exosomes released by quiescent SMB cells, activated SMB cell-derived exosomes contain more and significantly different protein components. The unique exosomal protein 'fingerprint' might reflect that exosomes from the SMB cells with different states of activation have distinguishing functional roles as well as targeting properties. Further bioinformatic analysis suggests that compared to quiescent exosomes, activated SMB cell-derived exosomes mainly participate in distinctive biological processes including activation of cellular component biogenesis and regulation of cell proliferation. Moreover, activated exosomes may function through the specific signaling pathways associated with the regulation of cell mitosis and immune response. Our results give valuable clues about the immunomodulatory functions of SMB cell-derived exosomes and provide the foundation for further investigation in the regulation mechanism of the adaptive immune response in lamprey.