, Volume 70, Issue 2, pp 831–842 | Cite as

Melilotus indicus extract induces apoptosis in hepatocellular carcinoma cells via a mechanism involving mitochondria-mediated pathways

  • Amer Ali Abd El-Hafeez
  • Hazim O. Khalifa
  • Rania Abdelrahman Elgawish
  • Samia A. Shouman
  • Magdy Hussein Abd El-Twab
  • Seiji Kawamoto
Original Article


Melilotus indicus, is a traditional medicine used as analgesic and emollient. Although Melilotus indicus extract (MIE) has recently been shown to suppress growth of several tumor cell lines, information regarding its antitumor mechanism is completely unknown. Here, we report the mechanism underlying the effects of MIE on human hepatocellular carcinoma cells, specifically HepG2, and SNU-182 cells. Methanolic MIE impaired the proliferation, and induced cell death in both HepG2 and SNU-182 cells but not in normal hepatic L-02 cells. Mechanistically, flow cytometric analysis revealed that MIE induces apoptosis in HepG2, and SNU-182 cells. However, MIE-induced apoptosis were not affected by a pan caspase inhibitor z-VAD-fmk as well as MIE did not stimulate caspase activation. Furthermore we found that MIE-induced apoptosis could be attributed to a mechanism involving mitochondria-mediated pathways evidenced by decrease in the mitochondrial membrane potential (ΔΨm), increase in the Bax/Bcl-2 ratio, and translocation of apoptosis inducing factor (AIF) from the mitochondria to the nucleus. Suppression in AIF expression by siRNA reduced MIE-induced apoptosis which suggested the dependency of MIE on AIF to induce apoptosis in hepatocellular carcinoma cells. To the best of our knowledge this is the first report elucidating the anticancer mechanism of MIE. Our findings suggested that MIE might be a good extract for developing anticancer drugs for human hepatocellular carcinoma treatment.


Melilotus indicus Cancer Apoptosis AIF 



This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflicts of interest.

Supplementary material

10616_2018_195_MOESM1_ESM.pptx (59 kb)
Supplementary material 1 (PPTX 58 kb)


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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Pharmacoloy and Experimental Oncology Unit, Cancer Biology Department, National Cancer InstituteCairo UniversityCairoEgypt
  2. 2.Hiroshima Research Center for Healthy Aging (HiHA), Department of Molecular Biotechnology, Graduate School of Advanced Sciences of MatterHiroshima UniversityHigashihiroshimaJapan
  3. 3.Department of Pharmacology, Faculty of Veterinary MedicineKafrelsheikh UniversityKafrelsheikhEgypt
  4. 4.Department of Forensic Medicine and Toxicology, Faculty of Veterinary MedicineSuez Canal UniversityIsmailiaEgypt
  5. 5.Graduate School of Biosphere ScienceHiroshima UniversityHigashihiroshimaJapan
  6. 6.Department of Botany and Microbiology, Faculty of ScienceMinia UniversityEl-Minia CityEgypt

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