Cuscuta campestris induces apoptosis by increasing reactive oxygen species generation in human leukemic cells

Document Type : Original Research Article


1 Golestan Rheumatology Research Center, Golestan University of Medical Sciences, Gorgan, Iran

2 Pharmacological Research Center of Medicinal Plants, Mashhad University of Medical Sciences, Mashhad, Iran

3 Division of Neurocognitive Sciences, Psychiatry and Behavioral Sciences Research Center, Mashhad University of Medical Sciences, Mashhad, Iran


Objective: Cuscuta campestris or common dodder is a holoparasitic  plant  that  has  been valorized for treatment of liver injury and cancer prevention in traditional medicine. Recently, extract of C. campestris had shown moderate antimicrobial properties and cytotoxic effects. In this study, we examined the level of cellular oxidants, cytotoxicity, apoptosis and differentiation induced by hydroalcoholic extract of C. campestris(CCE)(12.5-200 µg/ml), as well as arsenic trioxide (As2O3, 50 µM), in human leukemic (HL60 and NB4) and normal polymorph nuclear cells after 72 hr treatment.
Materials and Methods: Resazurin assay was used to determine cell viability following treatment with C. campestris. Intracellular reactive oxygen species (ROS) and apoptotic cells were measured by fluorimetry using carboxy 2′, 7′-dichlorofluorescein diacetate and propidium iodide (PI), as staining reagents, respectively. The differentiation of leukemic cells was evaluated by Giemsa staining and nitro blue tetrazolium (NBT) reduction.
Results: C. campestris inhibited cell viability with IC50 values of 23.9 µg/ml for HL60 and 60.3 µg/ml for NB4 cells after 72 hr treatment. ROS formation was also concentration-dependently increased following treatment with C. campestris. In addition, the number of apoptotic cells significantly increased to 88.4% and 62.3% in CCE (200 µg/ml)-treated HL60 and NB4 cells, respectively, which was higher than that of As2O3 (50 µM)-treated leukemic cells (p<0.001). Nonetheless, C. campestris did not induce differentiation of leukemic cells towards granulocytic pattern.
Conclusion: The present study demonstrated that C. campestris induced apoptosis through ROS production without having differential effect on leukemic cells, in concentration- and time-dependent manners. Understanding of precise signaling pathway by which C. campestris induce apoptosis, needs further research


Main Subjects

Alinejad B, Ghorbani A and Sadeghnia HR. 2013. Effects of combinations of curcumin, linalool, rutin, safranal, and thymoquinone on glucose/serum deprivation-induced cell death. Avicenna J Phytomed, 3: 321-328.
Benner SE and Hong WK. 1993. Clinical chemoprevention: developing a cancer prevention strategy. J Natl Cancer Inst, 85: 1446-1447.
Biswas S, Chowdhury A, Das J, Karamkar U, Raihan S and Das A. 2012. Phytochemical investigation and chromatographic evaluation with antimicrobial and cytotoxic potential of Cuscuta epithymum. Int J Pharm, 8: 422-427.
Candra E, Matsunaga K, Fujiwara H, Mimaki Y, Kuroda M, Sashida Y and Ohizumi Y. 2002. Potent apoptotic effects of saponins from Liliaceae plants in L1210 cells. J Pharm Pharmacol, 54: 257-262.
Cassatella M, Cappelli R, Della Bianca V, Grzeskowiak M, Dusi S and Berton G. 1988. Interferon-gamma activates human neutrophil oxygen metabolism and exocytosis. Immunology, 63: 499.
Choi HH, Jong H-S, Park J-H, Choi S, Lee JW, Kim T-Y, Otsuki T, Namba M and Bang Y-J. 2003. A novel ginseng saponin metabolite induces apoptosis and down-regulates fibroblast growth factor receptor 3 in myeloma cells. Int J Oncol, 23: 1087-1094.
Cragg GM and Newman DJ. 2005. Plants as a source of anti-cancer agents. J Ethnopharmacol, 100: 72-79.
Estañ MC, Calviño E, Calvo S, Guillén-Guío B, del Carmen Boyano-Adanez M, de Blas E, Rial E and Aller P. 2014. Apoptotic efficacy of etomoxir in human acute myeloid leukemia cells. Cooperation with arsenic trioxide and glycolytic inhibitors, and regulation by oxidative stress and protein kinase activities. PLoS One, 9: e115250.
Gaidi G, Correia M, Chauffert B, Beltramo J-L, Wagner H and Lacaille-Dubois M-A. 2002. Saponins-mediated potentiation of cisplatin accumulation and cytotoxicity in human colon cancer cells. Planta Med, 68: 70-72.
Ghazanfari T, Naseri M, Shams J and Rahmati B. 2013. Cytotoxic effects of Cuscuta extract on human cancer cell lines. Food Agric Immunol, 24: 87-94.
Gupta K, Stefan T, Ignatz-Hoover J, Moreton S, Parizher G, Saunthararajah Y and Wald DN. 2016. GSK-3 inhibition sensitizes acute myeloid leukemia cells to 1, 25D-mediated differentiation. Cancer Res, 76: 2743-2753.
Holm L. 1997. World weeds: natural histories and distribution, PP201-225. John Wiley & Sons.
Hosseini A and Rajabian A. 2016. Protective effect of Rheum turkestanikum root against doxorubicin-induced toxicity in H9c2 cells. Rev Bras Farmacogn, 26: 347-351.
Hu K and Yao X. 2003. The cytotoxicity of methyl protoneogracillin (NSC‐698793) and gracillin (NSC‐698787), two steroidal saponins from the rhizomes of Dioscorea collettii var. hypoglauca, against human cancer cells in vitro. Phytother Res, 17: 620-626.
Jafarian A, Ghannadi A and Mohebi B. 2014. Cytotoxic effects of chloroform and hydroalcoholic extracts of aerial parts of Cuscuta chinensis and Cuscuta epithymum on Hela, HT29 and MDA-MB-468 tumor cells. Res Pharm Sci, 9: 115.
Kajimoto S, Takanashi N, Kajimoto T, Xu M, Cao J, Masuda Y, Aiuchi T, Nakajo S, Ida Y and Nakaya K. 2002. Sophoranone, extracted from a traditional Chinese medicine Shan Dou Gen, induces apoptosis in human leukemia U937 cells via formation of reactive oxygen species and opening of mitochondrial permeability transition pores. Int J Cancer, 99: 879-890.
Mashkani B, Tanipour MH, Saadatmandzadeh M, Ashman LK and Griffith R. 2016. FMS-like tyrosine kinase 3 (FLT3) inhibitors: Molecular docking and experimental studies. Eur J Pharmacol, 776: 156-166.
McNeal JR, Arumugunathan K, Kuehl JV and Boore JL. 2007. Systematics and plastid genome evolution of the cryptically photosynthetic parasitic plant genus Cuscuta (Convolvulaceae). BMC Biol, 5: 1.
Mehrabani M, Modirian E, Ebrahimabadi A, Vafazadeh J, Shahnavaz S and Heidari MR. 2007. Study of the Effects of hydro-methanol extracts of lavandula vera DC. and cuscuta epithymum murr. on the seizure induced by pentylentetranzol in Mice. J Kerman Uni Med Sci, 14: 25-32.
Rodak BF, Fritsma GA and Keohane E. 2013. Hematology: clinical principles and applications, PP: 340-373. Elsevier Health Sciences.
Sadeghnia HR, Ghorbani Hesari T, Mortazavian SM, Mousavi SH, Tayarani-Najaran Z and Ghorbani A. 2014. Viola tricolor induces apoptosis in cancer cells and exhibits antiangiogenic activity on chicken chorioallantoic membrane. BioMed Res Inter, 2014.
Sak K. 2012. Chemotherapy and dietary phytochemical agents. Chemother Res Pract, 2012.
Sepehr MF, Jameie SB and Hajijafari B. 2011. The Cuscuta kotschyana effects on breast cancer cells line MCF7. J Med Plant Res, 5: 6344-6351.
Shafiee-Nick R, Ghorbani A, Vafaee Bagheri F and Rakhshandeh H. 2012. Chronic administration of a combination of six herbs inhibits the progression of hyperglycemia and decreases serum lipids and aspartate amino transferase activity in diabetic rats. Adv Pharmacol Sci, 2012.
Shen P, Wang S-L, Liu X-K, Yang C-R, Cai B and Yao X-S. 2003. Steroidal Saponins from Rhizomes of Tupistra wattii HOOK. f. Chem Pharm Bull (Tokyo), 51: 305-308.
Suganuma K, Miwa H, Imai N, Shikami M, Gotou M, Goto M, Mizuno S, Takahashi M, Yamamoto H and Hiramatsu A. 2010. Energy metabolism of leukemia cells: glycolysis versus oxidative phosphorylation. Leuk. Lymphoma, 51: 2112-2119.
Suresh V, Sruthi V, Padmaja B and Asha V. 2011. In vitro anti-inflammatory and anti-cancer activities of Cuscuta reflexa Roxb. J Ethnopharmacol, 134: 872-877.
Yan H, Peng Z-G, Wu Y-L, Jiang Y, Yu Y, Huang Y, Zhu Y-S, Zhao Q and Chen G-Q. 2005. Hypoxia-simulating agents and selective stimulation of arsenic trioxide-induced growth arrest and cell differentiation in acute promyelocytic leukemic cells. Haematologica, 90: 1607-1616.
Yokosuka A, Mimaki Y and Sashida Y. 2002. Spirostanol saponins from the rhizomes of Tacca chantrieri and their cytotoxic activity. Phytochemistry, 61: 73-78