Antiangiogenic Effects and Therapeutic Targets of Azadirachta indica Leaf Extract in Endothelial Cells

Department of Urology, Mayo Clinic/Foundation, Rochester, MN 55905, USA.
Evidence-based Complementary and Alternative Medicine (Impact Factor: 1.88). 02/2012; 2012:303019. DOI: 10.1155/2012/303019
Source: PubMed


Azadirachta indica (common name: neem) leaves have been found to possess immunomodulatory, anti-inflammatory and anti-carcinogenic properties. The present study evaluates anti-angiogenic potential of ethanol extract of neem leaves (EENL) in human umbilical vein endothelial cells (HUVECs). Treatment of HUVECs with EENL inhibited VEGF induced angiogenic response in vitro and in vivo. The in vitro proliferation, invasion and migration of HUVECs were suppressed with EENL. Nuclear fragmentation and abnormally small mitochondria with dilated cristae were observed in EENL treated HUVECs by transmission electron microscopy. Genome-wide mRNA expression profiling after treatment with EENL revealed differentially regulated genes. Expression changes of the genes were validated by quantitative real-time polymerase chain reaction. Additionally, increase in the expression of HMOX1, ATF3 and EGR1 proteins were determined by immunoblotting. Analysis of the compounds in the EENL by mass spectrometry suggests the presence of nimbolide, 2',3'-dehydrosalannol, 6-desacetyl nimbinene and nimolinone. We further confirmed antiproliferative activity of nimbolide and 2',3'-dehydrosalannol in HUVECs. Our results suggest that EENL by regulating the genes involved in cellular development and cell death functions could control cell proliferation, attenuate the stimulatory effects of VEGF and exert antiangiogenic effects. EENL treatment could have a potential therapeutic role during cancer progression.

Download full-text


Available from: Donald J Tindall,
    • "Surface antigen of B16 melanoma cell (B16MelSAg), breast tumor associated antigen (BTAA), and carcinoembryonic antigen (CEA) B16 melanoma tumor, breast cancer cells, and CEA + colorectal cancer cells [53] [55] [56] Alleviates mutagenicity of carcinogens Likely drug metabolizing enzymes In vivo bone marrow micronuclei test [55] [68] [71] Maintain cellular redox balance Antioxidant phase II enzymes, glutathione level, protein oxidation, and lipid oxidation and peroxidation Benzo(a)pyrene-induced stomach tumor model and DMBA-induced skin papilloma model, DMBA-induced rat mammary carcinogenesis model, MNNG-induced carcinogenesis model, and DMBA-induced HBP oral carcinogenesis model [72–76,85,87] Attenuates angiogenesis Human umbilical vein endothelial cells (HUVECs), vascular endothelial growth factor (VEGF) DMBA-induced HBP carcinogenesis model, chemical carcinogen-induced mammary tumorigenesis [13] [73] [89] 2. Azadirachtin Induces apoptosis Bcl-2 family proteins, survivin and caspase-3, -8, and -9 "
    [Show abstract] [Hide abstract]
    ABSTRACT: Azadirachta indica, also known as neem, is commonly found in many semi-tropical and tropical countries including India, Pakistan, and Bangladesh. The components extracted from neem plant have been used in traditional medicine for the cure of multiple diseases including cancer for centuries. The extracts of seeds, leaves, flowers, and fruits of neem have consistently shown chemopreventive and antitumor effects in different types of cancer. Azadirachtin and nimbolide are among the few bioactive components in neem that have been studied extensively, but research on a great number of additional bioactive components is warranted. The key anticancer effects of neem components on malignant cells include inhibition of cell proliferation, induction of cell death, suppression of cancer angiogenesis, restoration of cellular reduction/oxidation (redox) balance, and enhancement of the host immune responses against tumor cells. While the underlying mechanisms of these effects are mostly unclear, the suppression of NF-κB signaling pathway is, at least partially, involved in the anticancer functions of neem components. Importantly, the anti-proliferative and apoptosis-inducing effects of neem components are tumor selective as the effects on normal cells are significantly weaker. In addition, neem extracts sensitize cancer cells to immunotherapy and radiotherapy, and enhance the efficacy of certain cancer chemotherapeutic agents. This review summarizes the current updates on the anticancer effects of neem components and their possible impact on managing cancer incidence and treatment.
    Biochimica et Biophysica Acta (BBA) - Reviews on Cancer 08/2014; 1846(1):247-257. DOI:10.1016/j.bbcan.2014.07.002 · 7.85 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Neem (Azadirachta indica A. Juss), one of the most versatile medicinal plants that grows ubiquitously in India has attained worldwide prominence owing to its wide range of medicinal properties. The bioactivity of neem has been attributed to its rich content of complex limonoids. Neem extracts and limonoids have been documented to exert antiproliferative effects both in vitro and in vivo. Accumulating evidence indicates that the anticancer effects of neem extracts and neem limonoids are mediated by preventing carcinogen activation; enhancing host antioxidant and detoxification systems; inhibiting cell proliferation, inflammation, invasion and angiogenesis; inducing apoptosis; modulating oncogenic transcription factors and signalling kinases; and influencing the epigenome. Neem and its constituent limonoids that target multiple signalling pathways aberrant in cancer are promising candidates for anticancer drug development.
    Perspectives in Cancer Prevention-Translational Cancer Research, 01/2014: pages 45-60; , ISBN: 978-81-322-1532-5
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The present study was designed to gain insight into the antiproliferative activity of ethanolic neem leaves extract (ENLE) alone or in combination with cisplatin by cell viability assay on human breast (MCF-7) and cervical (HeLa) cancer cells. Nuclear morphological examination and cell cycle analysis were performed to determine the mode of cell death. Further, to identify its molecular targets, the expression of genes involved in apoptosis, cell cycle progression, and drug metabolism was analyzed by RT-PCR. Treatment of MCF-7, HeLa, and normal cells with ENLE differentially suppressed the growth of cancer cells in a dose- and time-dependent manner through apoptosis. Additionally, lower dose combinations of ENLE with cisplatin resulted in synergistic growth inhibition of these cells compared to the individual drugs (combination index <1). ENLE significantly modulated the expression of bax, cyclin D1, and cytochrome P450 monooxygenases (CYP 1A1 and CYP 1A2) in a time-dependent manner in these cells. Conclusively, these results emphasize the chemopreventive ability of neem alone or in combination with chemotherapeutic treatment to reduce the cytotoxic effects on normal cells, while potentiating their efficacy at lower doses. Thus, neem may be a prospective therapeutic agent to combat gynecological cancers.
    Journal of Oncology 01/2014; 2014(11):321754. DOI:10.1155/2014/321754
Show more