Gentiana macrophylla flavonoids from Tibetan medicine decreases circ_0059665 to alleviate the progression of non-small cell lung cancer under hypoxia.

BACKGROUND: Gentiana macrophylla Pall. is a traditional Tibetan medicinal herb possessing antinociceptive and anti-inflammatory activities. Circular RNAs (circRNAs) have been identified to be involved in the tumorigenesis of non-small cell lung cancer (NSCLC). Here, this study focused on investigating the function and mechanism of Gentiana macrophylla flavonoids (GF) and circ_0059665 in NSCLC progression. METHODS: The contents of mRNA and protein were detected using qRT-PCR and western blotting analysis. Cell proliferative and invasive abilities were evaluated by cell counting kit-8, EdU, colony formation and transwell assays, respectively. M2 macrophage polarization was analyzed by flow cytometry. RESULTS: GF treatment suppressed NSCLC cell proliferation, invasion and M2 macrophage polarization under hypoxic conditions. Circ_0059665 was highly expressed in NSCLC tissues and cells. Its expression was increased under hypoxic conditions but was reduced following GF treatment. Furthermore, circ_0059665 overexpression reversed the anticancer effects of GF on NSCLC cells under hypoxic conditions. Mechanistically, circ_0059665 acted as a sponge for miR-512-5p to regulate NOVA2 expression. Hypoxia decreased miR-512-5p levels, and increased NOVA2 levels in NSCLC cells, while these tendencies were abolished after GF treatment. Circ_0059665 silencing inhibited NSCLC cell proliferation, invasion and M2 macrophage polarization in hypoxic environments, which were counteracted by NOVA2 overexpression. Moreover, NOVA2 upregulation reversed the suppressive effects of GF on NSCLC cells with hypoxia treatment. In addition, GF impeded NSCLC tumor growth in vivo via suppressing circ_0059665. CONCLUSION: GF treatment in hypoxic environments suppressed NSCLC cell proliferation, invasion and M2 macrophage polarization via the circ_0059665/miR-512-5p/NOVA2 axis.

The complete chloroplast genome sequence of Heracleum millefolium Diels (Apiaceae).

We assembled the complete chloroplast genome of Heracleum millefolium which is a traditional widely used medicinal plant in China. The whole genome is 150,025 bp in length which was divided into four subregions: a large single-copy region (93,645 bp), a pair of 19,458 bp inverted repeats regions, and a small single-copy region (17,464 bp), respectively. Additionally, the chloroplast genome of H. millefolium detected 128 genes, including 85 protein coding genes, 36 transfer RNAs, and eight ribosomal RNAs. The overall GC content of this chloroplast genome is 37.5% and the mean coverage value is 1752.4x. Phylogenetic analysis based on 17 chloroplast genomes dataset was conducted to clarify the relationships of the major clades in Apiaceae. The results strongly supported the monophyly of Heracleum and the closer relationship of H. millefolium and H. candicans.

Determination of volatile components in Tibetan medicine Heracleum millefolium by HS-GC-MS / 中草药

Objective: To establish a rapid analytical method for volatile components in Tibetan medicine Heracleum millefolium and to determine the volatile components from its roots, stems, leaves, and flower parts, respectively. Methods: Headspace sampling incorporation with gas-chromatography-mass spectrum (HS-GC-MS) determination was introduced to analyze the powder directly. Static headspace equilibration was performed at 100℃ for 40 min, and 1 mL of the headspace gas was injected in split mode of 10:1. The split inlet temperature was 260℃. The carrier gas was He at a constant flow rate of 1.8 mL/min. The column oven temperature was initially set at 50℃ for 2 min, then increased to 100℃ at 2℃/min, held for 6 min, then increased to 300℃ at 10℃/min and held for 2 min. The GC/MS interface temperature was maintained at 280℃. The solvent delay time was 3 min (to bypass the solvent peak). The volatile components were confirmed by NIST11.L database, and volatile organic compounds from roots, stems, leaves, and flower parts were compared. Results: The types of compounds in the roots, stems, leaves, and flowers of H. millefolium are mainly aldehyde, benzene, alcohols, and alkene. Octanal, hexanal, and γ-terpinene are the main components in the roots and stems. While o-isopropyltoluene and terpinolene are the main components in the leaves and flowers. Conclusion: HS-GC-MS method is easy, simple, and feasible, and can be widely used in other Chinese materia medica samples for analysis of volatile components.