9-Hydroxycanthin-6-one isolated from stem bark of Ailanthus altissima induces ovarian cancer cell apoptosis and inhibits the activation of tumor-associated macrophages.

The stem bark of Ailanthus altissima is used in traditional medicine in Asia to treat a variety of diseases, including cancer. The aim of this study was to identify compounds with tumoricidal activity from A. altissima stem bark and to investigate their mechanisms of action. Among the 13 compounds isolated from the ethyl acetate fraction of A. altissima stem bark, the ß-carboline alkaloid 9-hydroxycanthin-6-one had potent cytotoxicity in all three ovarian cancer cell types examined. 9-Hydroxycanthin-6-one induced apoptosis through the activation of caspases-3, -8, and -9. 9-Hydroxycanthin-6-one increased the intracellular levels of reactive oxygen species (ROS), and pre-treatment with the antioxidant N-acetyl-l-cysteine (NAC) attenuated the pro-apoptotic activity of 9-hydroxycanthin-6-one. Additionally, 9-hydroxycanthin-6-one was found to decrease the expressions of MCP-1 and RANTES, major determinants of macrophage recruitment at tumor sites, in ovarian cancer cells. Treatment with 9-hydroxycanthin-6-one inhibited the levels of M2 phenotype markers and some cancer-promoting factors, such as MMP-2, MMP-9, and VEGF, in macrophages educated in ovarian cancer conditioned medium. Taken together, these data suggest that 9-hydroxycanthin-6-one isolated from A. altissima stem bark induces apoptosis in human ovarian cancer cells through the caspase- and ROS-dependent pathways and inhibits the activation of tumor-associated macrophages.

Photocatalytic degradation of synthetic food dye, sunset yellow FCF (FD&C yellow no. 6) by Ailanthus excelsa Roxb. possessing antioxidant and cytotoxic activity.

The purpose of our work is to identify the bioactive compounds of bark and leaves extract from Ailanthus excelsa Roxb. and to explore its effectiveness against synthetic food dye. The presence of primary and secondary metabolites was confirmed by carrying out phytochemicals analysis. With the prior knowledge accessible on the indispensable secondary metabolites holding antioxidant and cytotoxicity activity, the quantitative screening of total phenolic and flavonoid content in methanolic and aqueous extract of bark and leaves from Ailanthus excelsa were done. Comparatively, a higher value of flavonoid (161±0.3µg/mg) and phenolic acid content (152.4±0.14µg/mg) was found in bark extract. By FTIR analysis, the characteristic peak was obtained at 1581.63 and 1598.99cm-1 confirmed the presence of functional groups associated to flavonoids and other phenolic groups respectively. In bark extract, 81% of DPPH inhibition was observed when compared to ascorbic acid (standard) 92% of free radical scavenging activity. Bark extract from Ailanthus excelsa exhibited 71% cytotoxicity against HeLa cell line (cervical cancer). In examining the toxicity level of crude extracts with red blood cells (RBC), the bark extract was showed a very less (2.8%) haemolytic activity. They also showed maximum zone of inhibition in antibacterial activity i.e. 13±0.5mm against Escherichia coli culture. At a concentration of 10mg/mL of crude extract from A. excelsa, 55% degradation of sunset yellow dye was observed. It concludes that, the compounds present in the A. excelsa, especially the bark extract showed better photocatalytic, haemolytic, antioxidant, cytotoxicity and antibacterial activity when compared to leaves extract.

Ailanthone induces G2/M cell cycle arrest and apoptosis of SGC­7901 human gastric cancer cells.

Ailanthone is a major quassinoid extracted from the Chinese medicinal herb Ailanthus altissima, which has been reported to exert antiproliferative effects on various cancer cells. The present study aimed to investigate the antitumor effects of ailanthone on SGC­7901 cells, and to analyze its underlying molecular mechanisms. Following treatment with ailanthone, Cell Counting kit­8 was used to detect the cytotoxic effects of ailanthone on SGC­7901 cells in vitro. The typical apoptotic morphology of SGC­7901 cells was observed by Hoechst 33258 staining. Cell cycle progression and apoptosis were measured by flow cytometry, and the protein and mRNA expression levels of Bcl­2 and Bax were analyzed by western blot analysis and reverse transcription­quantitative polymerase chain reaction (RT­qPCR) respectively, in SGC­7901 cells. The results of the present study indicated that ailanthone inhibited the proliferation of SGC­7901 cells in a dose­ and time­dependent manner in vitro, and also demonstrated that ailanthone induced G2/M phase cell cycle arrest and apoptosis of SGC­7901 cells. Furthermore, analysis of the underlying molecular mechanisms revealed that ailanthone downregulated the expression levels of Bcl­2, whereas the expression levels of Bax were upregulated at the protein and mRNA levels. In conclusion, ailanthone may inhibit the proliferation of SGC­7901 cells by inducing G2/M phase cell cycle arrest and apoptosis via altering the protein and mRNA expression levels of Bcl­2 and Bax in SGC­7901 cells.

Ailanthone Inhibits Huh7 Cancer Cell Growth via Cell Cycle Arrest and Apoptosis In Vitro and In Vivo.

While searching for natural anti-hepatocellular carcinoma (HCC) components in Ailanthus altissima, we discovered that ailanthone had potent antineoplastic activity against HCC. However, the molecular mechanisms underlying the antitumor effect of ailanthone on HCC have not been examined. In this study, the antitumor activity and the underlying mechanisms of ailanthone were evaluated in vitro and in vivo. Mechanistic studies showed that ailanthone induced G0/G1-phase cell cycle arrest, as indicated by decreased expression of cyclins and CDKs and increased expression of p21 and p27. Our results demonstrated that ailanthone triggered DNA damage characterized by activation of the ATM/ATR pathway. Moreover, ailanthone-induced cell death was associated with apoptosis, as evidenced by an increased ratio of cells in the subG1 phase and by PARP cleavage and caspase activation. Ailanthone-induced apoptosis was mitochondrion-mediated and involved the PI3K/AKT signaling pathway in Huh7 cells. In vivo studies demonstrated that ailanthone inhibited the growth and angiogenesis of tumor xenografts without significant secondary adverse effects, indicating its safety for treating HCC. In conclusion, our study is the first to report the efficacy of ailanthone against Huh7 cells and to elucidate its underlying molecular mechanisms. These findings suggest that ailanthone is a potential agent for the treatment of liver cancer.

The effects of iron deficiency on lead accumulation in Ailanthus altissima (Mill.) Swingle seedlings.

Understanding the effects of root Fe deficiency on the uptake and translocation of toxic metals can be important for improving the phytoremediation strategies of polluted soils. The present study investigated how Fe nutritional status affects the uptake and root-to-shoot-translocation of Pb in hydroponically grown seedlings of (Mill.) Swingle. The interactions of Fe deficiency and Pb were assessed by measuring the root Fe(III) reductase activity, carboxylic acids concentration in root exudates, root and shoot biomass, and accumulation of Pb and other metals (Fe, Zn, Mn, and Cu) in roots and leaves of Fe-sufficient (+Fe) and Fe-deficient (-Fe) plants. The results indicate that Fe deficiency induced 18-fold higher Fe(III) reductase activity in roots compared with +Fe plants, which was followed by increased root exudation of citric acid (28.2 ± 1.39 in +Fe and 498 ± 256.4 µmol g DW 2 h in -Fe plants). Iron deficiency also induces a significant decrease of root and shoot dry weight compared with the control +Fe plants, whereas 2-wk Pb (20 µM) treatment did not influence root and shoot growth. Iron-sufficient plants accumulated more Pb (56.8 ± 17.29 µg g) in leaves than -Fe plants (21.5 ± 8.10 µg g). Two weeks of exposure to Pb significantly decreased Fe(III) reductase activity and accumulation of Fe, Zn, and Mn in the roots of -Fe plants. It is hypothesized that 2 wk of root exposure to Pb blocks functioning of a specific Fe transport system activated under Fe deficiency.