Neferine, an alkaloid ingredient in lotus seed embryo, inhibits proliferation of human osteosarcoma cells by promoting p38 MAPK-mediated p21 stabilization

Key Laboratory of Experimental Teratology, Ministry of Education, Institute of Molecular Medicine and Genetics, Shandong University School of Medicine, Jinan, Shandong 250012, China.
European journal of pharmacology (Impact Factor: 2.53). 02/2012; 677(1-3):47-54. DOI: 10.1016/j.ejphar.2011.12.035
Source: PubMed


Identification of natural products that have antitumor activity is invaluable to the chemoprevention and therapy of cancer. The embryos of lotus (Nelumbo nucifera) seeds are consumed in beverage in some parts of the world for their presumed health-benefiting effects. In this report we studied the effects of neferine, a major alkaloid component in lotus embryos, on human osteosarcoma cells and the underlying mechanisms. We found that neferine possessed a potent growth-inhibitory effect on human osteosarcoma cells, but not on non-neoplastic human osteoblast cells. The inhibitory effect of neferine on human osteosarcoma cells was largely attributed to cell cycle arrest at G1. The induction of G1 arrest was p21(WAF1/CIP1)-dependent, but was independent of p53 or RB (retinoblastoma-associated protein). The up-regulation of p21 by neferine was due to an increase in the half-life of p21 protein. We examined four kinases that are known to affect the stabilization of p21, and found that p38 MAPK and JNK were activated by neferine. However, only SB203580 (an inhibitor of p38), but not SP600125 (the inhibitor of JNK), can attenuate the up-regulation of p21 in response to neferine. Furthermore, the p21-stabilizing effect of neferine was abolished when p38 was silenced by RNA interference. Finally, we showed that neferine treatment led to an increased phosphorylation of p21 at Ser130 that was dependent on p38. Our results for the first time showed a direct antitumor effect of neferine, suggesting that consumption of neferine may have cancer-preventive and cancer-therapeutic benefit.

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    • "Jasminum sambac: Flower beta-primeveroside, 2-phenylethyl-beta primeveroside, beta-rutinoside, linalool, methylanthranilate (Inagaki et al., 1995) Flower 4-hexanolide, 4-nonanolide, 2-hexenyl-hexanoate, 4-hydroxy-2,5-diethyl-3(2H) furanone (Ito et al., 2002) Flower cis-3-hexenyl-acetate, (E)-beta-ocimene, linalool, benzylacetate, (E,E)-alpha-farnesene (Pragadheesh et al., 2011) flower coumarins, cardiac glycosides, essential oils, flavonoids, phenolics, saponins, steroids (Kunhachan et al., 2012) Flower linalool, benzyl-acetate, a-farnesene (Kanlayavattanakul et al., 2013) Mammea siamensis: Flower mammea B/AC cyclo D4-alkylcoumarin; mammea A/AC cyclo D4-phenylcoumarin (Kaweetripob et al., 2000) Flower mammea A/AA cyclo D, mammea A/AD cyclo D, mammea A/AB cyclo D, mammea A/AC cyclo F, mammea A/AB cyclo F, mammea A/AA cyclo F, mammea B/AC cyclo F (Prachyawarakorn et al., 2000) Flower mammea E/BA cyclo D, mammea E/BC cyclo D, mammea E/D cyclo BD, mammea E/AC cyclo D (Mahidol et al., 2002) Flower mammeasin A and B, suragins B and D, kayeassamins E,F and G (Morikawa et al., 2012) Flower mammeanoyl (Tung et al., 2013) Mesua ferrea: Flower 4-alkyl and 4-phenyl 5,7-dihydroxycoumarins (Verotta et al., 2004) Michela alba: Flower alpha-myrcene, (S)-limonene, (R)-fenchone, linalool, camphor, caryophyllene, germacrene D (Shang et al., 2002) Flower linalool (Xia et al., 2010) Mimusops elengi: Flowers beta-sitosterol, beta-sitosterol glycosides, quercitol, ursolic acid, lupeol (Gami et al., 2012) Nelumbo nucifera: Flower myricetin, quercetin, kaempferol, isorhamnetin, diosmetin, syringetin (Chen et al., 2012) Flower N-methylasimilobine N-oxide, 2-hydroxy-1-methoxy-6a, 7-dehydroaporphine (Nakamura et al., 2013) Stamens myricetin, quercetin, kaempferol, isorhamnetin, diosmetin, syringetin, anthoxyanins (Chen et al., 2013) Stamens kaempherol glycosides, beta-sitosterol glycoside (Jung et al., 2003) Stamens kaempferol (Shim et al., 2009) Petals syringetin glucoside, quercetin glucoside, isorhamnetin glucoside, kaempferol glucoside (Xingfeng et al., 2010) Petals myricetin, quercetin, kaempferol, isorhamnetin, diosmetin, syringetin, anthoxyanins (Chen et al., 2013) Petals malvidin 3-O-glucoside (Deng et al., 2013) Pistills myricetin, quercetin, kaempferol, isorhamnetin, diosmetin, syringetin, anthoxyanins (Chen et al., 2013) Receptacles hyperoside glycosides, isoquercitrin glycosides, quercetin glycosides, isorhamnetin glycosides, syringetin glycosides (Wu et al., 2012; Wu et al., 2013) pathway activation (Zhang et al., 2012; Poornima et al., 2013a, 2013b, 2014; Yoon et al., 2013). Liensinine, neferine, and isoliensinine are substrates. "
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    ABSTRACT: Ethnopharmacological relevance: Thai medicine has a long tradition of tonifying medicinal plants. In the present investigation, we studied the flower extracts of Jasminum sambac, Mammea siamensis, Mesua ferrea, Michelia alba, Mimusops elengi, and Nelumbo nucifera and speculated that these plants might influence metabolism and substance flow in the body. Materials and methods: Isolation of porcine brain capillary endothelial cells (PBCECs) as well as multidrug-resistance CEM/ADR5000 leukemia cells, MDA-M;B-231 breast cancer, U-251 brain tumor, and HCT-116 colon cancer cells were used. The calcein-acetoxymethylester (AM) assay was used to measure inhibition of P-glycoprotein transport. XTT and resazurin assays served for measuring cytotoxicity. Results: The extracts revealed cytotoxicity towards CCRF-CEM leukemia cells to a different extent. The strongest growth inhibition was found for the n-hexane extracts of Mammea siamensis and Mesua ferrea, and the dichloromethane extracts of Mesua ferrea and Michelia alba. The flower extracts also inhibited P-glycoprotein function in porcine brain capillary endothelial cells and CEM/ADR5000 leukemia cells, indicating modulation of the blood-brain barrier and multidrug resistance of tumors. Bioactivity-guided isolation of coumarins from Mammea siamensis flowers revealed considerable cytotoxicity of mammea A/AA, deacetylmammea E/BA and deacetylmammea E/BB towards human MDA-MB-231 breast cancer, U-251 brain tumor, HCT-116 colon cancer, and CCRF-CEM leukemia cells. Conclusion: The plants analyzed may be valuable in developing novel treatment strategies to overcome the blood-brain barrier and multidrug-resistance in tumor cells mediated by P-glycoprotein.
    Full-text · Article · Jun 2014 · Journal of ethnopharmacology
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    • "Recently, we have reported that neferine could induce ROS dependent mitochondrial mediated apoptosis in liver and lung cancer cells (Poornima et al., 2013a, 2013b). Zhang et al. (2012) showed that neferine could inhibit the proliferation of osteosarcoma cells. It could reverse the multidrug resistance in MCF-7/ ADM, HepG2 (Cheng et al., 2008) and human gastric carcinoma cells (Cao et al., 2004). "
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    ABSTRACT: Doxorubicin (DOX) is the best anticancer agent that has ever been used, but acquired tumor resistance and dose limiting toxicity are major road blocks. Concomitant use of natural compounds is a promising strategy to overcome this problem. Neferine, a proven anticancer-agent is found in green embryos of lotus seed. The study demonstrates that neferine acts as an effective enhancer of DOX-induced cell death in A549 cells through ROS mediated apoptosis with MAPK activation and inhibition of NF-κB nuclear translocation. Cotreatment of cells with neferine significantly enhanced intracellular DOX-accumulation. Neferine and DOX in combination also triggered oxidative stress through intracellular Ca2+ accumulation and dissipation of mitochondrial membrane potential in addition to significant loss of cellular antioxidant pool. The MAPK inhibitor effectively decreased the cell-death induced by neferine and DOX. Pretreatment of cells with glutathione reversed the apoptosis induced by the combined regimen and recovered the Bcl2/Bax ratio. Moreover, neferine treatment significantly increased the cell viability of DOX-treated cardiomyocytes indicating a possible protective role of neferine towards DOX-induced cardiotoxicity. Taken together, our results suggest that a strategy of using neferine and DOX in combination could be helpful to increase the efficacy of DOX and to achieve anticancer synergism by curbing the toxicity.
    Full-text · Article · Mar 2014 · Food and chemical toxicology: an international journal published for the British Industrial Biological Research Association
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    • "Previous studies have shown that neferine has a number of pharmacological actions and is useful in the treatment of high fevers and hyposomnia, arrhythmia , platelet aggregation, thrombosis, obesity, and in the inhibition of pulmonary fibrosis [8]. In recent years, it has been reported that neferine inhibits the proliferation of vascular smooth muscle cells [9], hypertrophic scar fibroblasts [10], and osteosarcoma cells [11]. A pharmacokinetic study showed that the distribution of neferine to the lung is the highest of all the tissues after an oral administration of 50 mg/kg of neferine to rats [12]. "
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    ABSTRACT: Neferine is the major bisbenzylisoquinoline alkaloid isolated from the seed embryo of a traditional medicinal plant Nelumbo nucifera (Lotus). Epidemiological studies have revealed the therapeutic potential of lotus seed embryo. Although several mechanisms have been proposed, a clear anticancer action mechanism of neferine on lung cancer cells is still not known. Lung cancer is the most common cause of cancer death in the world, and the patients with advanced stage of nonsmall lung cancer require adjunct chemotherapy after surgical resection for the eradication of cancer cells. In this study, the effects of neferine were evaluated and characterized in A549 cells. Neferine induced apoptosis in a dose-dependent manner with the hypergeneration of reactive oxygen species, activation of MAPKs, lipid peroxidation, depletion of cellular antioxidant pool, loss of mitochondrial membrane potential, and intracellular calcium accumulation. Furthermore, neferine treatment leads to the inhibition of nuclear factor kappaB and Bcl2, upregulation of Bax and Bad, release of cytochrome C, activation of caspase cascade, and DNA fragmentation. In addition, neferine could induce p53 and its effector protein p21 and downregulation of cell cycle regulatory protein cyclin D1 thereby inducing G1 cell cycle arrest. These results suggest a novel function of neferine as an apoptosis inducer in lung cancer cells. © 2013 BioFactors, 2013.
    Full-text · Article · Feb 2014 · BioFactors
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