Harmine derivative B-9-3 inhibits non-small cell lung cancer via the VEGFA/PI3K/AKT pathway

BackgroundThis study aimed to investigate the molecular mechanism by which the Harmine derivative B-9-3 inhibits angiogenesis and promotes apoptosis in non-small cell lung cancer (NSCLC).MethodsThree non-small cell lung cancer (NSCLC) models (human NSCLC cell line A549, human lung squamous cell carcinoma cell line H226, human large cell lung carcinoma cell line H460) were established. Cell proliferation was assessed using CCK-8 assays and colony formation assays. Cell motility was evaluated through scratch wound healing, invasion, and migration assays. Cell apoptosis was analyzed by Hoechst 33258 staining, AO/EB fluorescence staining, and flow cytometry. Real-time PCR was used to measure the mRNA expression of B-cell lymphoma/leukemia-2 (Bcl-2), Bcl-2-associated X protein (Bax), and Caspase-3, while Western blotting was performed to assess the protein levels of vascular endothelial growth factor A (VEGFA), phosphatidylinositol 3-kinases p110 Beta (PI3K), phospho-phosphatidylinositol 3-kinases (p-PI3K), protein kinase B (AKT), phosphorylated protein kinase B (p-AKT), Bax, Bcl-2, and Caspase-3.ResultsCompared to the control group, B-9-3 (50, 100, 200 μg/mL) inhibited the growth and motility of the three types of lung cancer cells, suppressed cell invasion and migration, and promoted cell apoptosis and necrosis. The apoptosis rates in three types of non-small cell lung cancer (NSCLC) cells were significantly increased. The mRNA expressions of Bax and Caspase-3 were markedly upregulated, while that of Bcl-2 was significantly downregulated. Additionally, the protein levels of VEGFA, p-PI3K/PI3K, p-AKT/AKT, and Bcl-2 were notably reduced, whereas the protein levels of Bax and Caspase-3 were significantly elevated.ConclusionThe harmine derivative B-9-3 may exert its anti-NSCLC effects by inhibiting angiogenesis and promoting lung cancer cell apoptosis via the VEGFA/PI3K/AKT signaling pathway

Effect of 17α-methyltestosterone (MT) on osmoregulatory responses and apoptosis in genetically improved farmed tilapia (GIFT), Oreochromis niloticus (L.)

Androgenic compounds can affect osmoregulatory response and apoptosis in fish. In the present study, we exposed genetically improved farmed tilapia (GIFT) Oreochromis niloticus (L.) to 17α-methyltestosterone (MT, 0.5 and 5 mg/L) for 7, 14 and 21 days for understanding the phenomenon. The activities of Na+/K+ ATPase (NKA) and Ca2+/Mg2+ ATPase (CMA) were measured in the gill, kidney and intestine to evaluate the change in osmoregulation of GIFT, and genotoxicity was also detected. Results showed that organic NKA were significantly decreased in 5 mg/L MT exposure groups. The intestine NKA was significantly increased (0.5 mg/L MT). MT exposures increased the CMA of kidney and intestine (0.5 mg/L), together with gill CMA (5 mg/L MT). The results of genotoxicity assay showed gill atp1a1a and nkcc2 transcripts significantly increased, while intestine atp1a1a and fxyd7 transcripts revealed significant increases for MT exposure groups. Caspases proteins demonstrated significant increases at 7th and 21st day, and their transcripts were enhanced in 0.5 mg/L MT exposure groups. The results have evidently demonstrated that chronic exposure of MT could result in organic osmoregulatory response and hepatic apoptosis in GIFT O. niloticus

Effect of 17α-methyltestosterone (MT) on osmoregulatory responses and apoptosis in genetically improved farmed tilapia (GIFT), Oreochromis niloticus (L.)

361-368Androgenic compounds can affect osmoregulatory response and apoptosis in fish. In the present study, we exposed genetically improved farmed tilapia (GIFT) Oreochromis niloticus (L.) to 17α-methyltestosterone (MT, 0.5 and 5 mg/L) for 7, 14 and 21 days for understanding the phenomenon. The activities of Na+/K+ ATPase (NKA) and Ca2+/Mg2+ ATPase (CMA) were measured in the gill, kidney and intestine to evaluate the change in osmoregulation of GIFT, and genotoxicity was also detected. Results showed that organic NKA were significantly decreased in 5 mg/L MT exposure groups. The intestine NKA was significantly increased (0.5 mg/L MT). MT exposures increased the CMA of kidney and intestine (0.5 mg/L), together with gill CMA (5 mg/L MT). The results of genotoxicity assay showed gill atp1a1a and nkcc2 transcripts significantly increased, while intestine atp1a1a and fxyd7 transcripts revealed significant increases for MT exposure groups. Caspases proteins demonstrated significant increases at 7th and 21st day, and their transcripts were enhanced in 0.5 mg/L MT exposure groups. The results have evidently demonstrated that chronic exposure of MT could result in organic osmoregulatory response and hepatic apoptosis in GIFT O. niloticus

A Chromosome-Level Genome Assembly of the Mandarin Fish (Siniperca chuatsi)

The mandarin fish, Siniperca chuatsi, is an economically important perciform species with widespread aquaculture practices in China. Its special feeding habit, acceptance of only live prey fishes, contributes to its delicious meat. However, little is currently known about related genetic mechanisms. Here, we performed whole-genome sequencing and assembled a 758.78 Mb genome assembly of the mandarin fish, with the scaffold and contig N50 values reaching 2.64 Mb and 46.11 Kb, respectively. Approximately 92.8% of the scaffolds were ordered onto 24 chromosomes (Chrs) with the assistance of a previously established genetic linkage map. The chromosome-level genome contained 19,904 protein-coding genes, of which 19,059 (95.75%) genes were functionally annotated. The special feeding behavior of mandarin fish could be attributable to the interaction of a variety of sense organs (such as vision, smell, and endocrine organs). Through comparative genomics analysis, some interesting results were found. For example, olfactory receptor (OR) genes (especially the beta and delta types) underwent a significant expansion, and endocrinology/vision related npy, spexin, and opsin genes presented various functional mutations. These may contribute to the special feeding habit of the mandarin fish by strengthening the olfactory and visual systems. Meanwhile, previously identified sex-related genes and quantitative trait locis (QTLs) were localized on the Chr14 and Chr17, respectively. 155 toxin proteins were predicted from mandarin fish genome. In summary, the high-quality genome assembly of the mandarin fish provides novel insights into the feeding habit of live prey and offers a valuable genetic resource for the quality improvement of this freshwater fish

Characterization of the growth-related transcriptome in the Liver and Brain of mandarin fish (Siniperca chuatsi) through RNA-Seq analysis

The brain and liver of mandarin fish (Siniperca chuatsi) produce essential hormones for their growth and development. However, the mechanisms underlying the interplay of these hormones remain unclear. This study aimed to assess the slow – and fast-growing traits, including body weight and length of mandarin fish reared in a single pond. To compare the differentially expressed genes (DEGs) in slow – and fast-growing fishes, we performed RNA sequencing analysis. After assembling the clean reads, 192,280 transcripts and 100,416 unigenes were obtained. 344 and 845 DEGs in the brain and liver in the fast-and slow-growing group were annotated respectively. These DEGs were associated with signaling pathways involved in growth and development, such as AGE-RAGE, Transforming growth factor-β/Smad, Mitogen-activated protein kinase, Wingless/Integrated, and Insulin-like growth factor (IGF) 1. Furthermore, our fasting experiment revealed a concurrent decrease in body weight and the expression levels of genes related to the growth hormone (GH)/IGF system, alongside an increase in the insulin-like growth factor binding protein gene IGFBP3 expression. These findings suggest the GH/IGF system regulates body weight during fasting periods. The vast amount of transcriptome data generated in this study substantially expands the existing gene and genome resources for the growth traits of mandarin fish

Molecular characterization and expression profile of the estrogen receptor α gene during different reproductive phases in Monopterus albus

AbstractTo understand the molecular mechanism of estrogen and to evaluate the role of the estrogen receptor in mediating estrogen action, the full-length cDNA of estrogen receptor α (ERα) was cloned from Monopterus albus and its expression pattern and distribution were investigated. The ERα cDNA of M. albus includes an open reading frame of 1863 bp, a 140-bp 5’-untranslated region and a 797-bp 3’-untranslated region. Amino acid sequence homology analysis showed that the Monopterus albus ERα has a moderate degree of similarity with Sebastes schlegelii, Zoarces viviparus and Haplochromis burtoni (81.1%, 80.7% and 80.4%, respectively). Quantitative PCR results showed that the highest level of ERα expression was in the liver; the next highest level of expression was observed in the gonads, where it was expressed at high levels particularly in the ovary in developmental stages IV and V and in the testis in developmental stage II/III. Immunohistochemistry analysis showed that ERα was present as slender particles distributed mainly in the membranes of spermatocytes and oocytes in the testis and ovary, whereas no positive signal was observed in the cytoplasm of sperm cells. This report describes the first molecular characterization of full-length ERα and its tissue-specific distribution in M. albus.</jats:p