In collaboration with Payame Noor University and Iranian Society of Physiology and Pharmacology

Document Type : Article

Author

1 Assistant Professor, Department of Biology, Faculty of ‎Science, Shahid Chamran University of Ahvaz, ‎Ahvaz, Iran

2 Stem cells and transgenic technology research ‎center (STTRC), Shahid Chamran University of ‎Ahvaz, Ahvaz, Iran

Abstract

Lung cancer is one the most common cancer in the world and Iran that lead to many deaths.  Despite abundant researches, an appropriate cure has not been yet found for cancer. Citrus extract contains abundant anti-oxidative compounds. In this research evaluated and compared the toxic effect of Citrus leaves hydroalcoholic extracts on the proliferation and viability of A459 cancer cells. Citrus leaves extracts were prepared by soxhlet set. A549 cells were proliferated and treated with 0.05-10 mg/ml concentrations of extracts for 72h. Cell viability and morphology were evaluated by MTT assay and acridine orange staining on certain times after treatment. Comparison of the cell viability percent in experimental groups indicated that IC50 were 2, 6, and 8 respectively for Citrus limon, Citrus reticulata, and Citrus aurantium leaves extract (P<0.05). Also, number of apoptotic cells was significantly (P<0.05) more in C.lemon group than C. reticulata and C. aurantium group and indeed, number of apoptotic cells were significantly (P<0.05) more in C. reticulata group than C. aurantium group. Citrus leaves hydroalcoholic extract can induce apoptosis in A549 cells in a dose and time-dependent manner.

Keywords

Ademosun, A.O.; Oboh, G.; Passamonti, S.; Tramer, F.; Ziberna, L.; Boligon, A.A.; Athayde, M.L. (‎‎(2015. Inhibition of metalloproteinase and proteasome activities in colon cancer ‎cells by citrus peel ‎extracts. J Basic Clin Physiol Pharmacol; 26(5): 471-477.‎
Ahn, K.I.; Choi, E.O.; Kwon, D.H.; HwangBo, H.; Kim, M.Y.; Kim, H.J.; Ji, S.Y.; Hong, S.H.; Jeong, J.‎W.; ‎Park, C.; Kim, N.D.; Kim, W.J.; Choi, Y.H. (2017). Induction of apoptosis by ‎ethanol extract of Citrus ‎unshiu Markovich peel in human bladder cancer T24 cells through ‎ROS-mediated inactivation of the ‎PI3K/Akt pathway. Biosci Trends; 11(5): 565-573.‎
Azanchi, T.; Shafaroodi, H.; Asgarpanah, J. (2014). Anticonvulsant activity of Citrus ‎aurantium ‎blossom essential oil (neroli): involvment of the GABAergic system. Nat Prod ‎Commun; 9(11): 1615-‎‎1618.‎
Boeing, J.S.; Barizao, E.O.; BC, E.S.; Montanher, P.F.; de Cinque Almeida, V.; ‎Visentainer, J.V. (2014). ‎Evaluation of solvent effect on the extraction of phenolic compounds and ‎antioxidant capacities from ‎the berries: application of principal component analysis. Chem ‎Cent J; 8 (1):48.‎
Bonner, M.Y.; Arbiser, J.L. (2014). The antioxidant paradox: what are antioxidants and how ‎should ‎they be used in a therapeutic context for cancer. Future Med Chem; 6(12): 1413-1422.‏
Chiang, S.Y.; Kim, S.M.; Kim, C.; Um, J.Y.; Park, K.R.; Kim, S.W.; Lee, S.G.; Jang, H.J.; ‎Nam, D.; Ahn, ‎ K.S.; Kim, S.H.; Choi, S.H.; Shim, B.S.; Na, Y.C.; Jeong, E.K.; Cho, S.K.; (2012). Antiproliferative effects of ‎Dangyuja (Citrus grandis Osbeck) leaves through ‎suppression of constitutive signal transducer and ‎activator of transcription 3 activation in ‎human prostate carcinoma DU145 cells. J Med Food; 15‎‎(2): 152-160.‎
Dehpour Jobari, A.; Hasabpour, Z. (2016). Cytotoxicity Effects of Citrus Skin Extrct on Cancer Cell line MCF-7. J of Animal Biology; 8(2): 19-25.
Jafarpour, M.; Yousefi, G.; Hamedi, A.; Shariat, A.; Salehi, A.; Heydari, M. (2016). ‏Effect of a ‎traditional ‎syrup from Citrus medica L. fruit juice on migraine headache: A randomized ‎double blind placebo ‎controlled clinical trial. J Ethnopharmacol; 179: 170-176.‎
Jaganathan, S.K.; Vellayappan, M.V.; Narasimhan, G.; Supriyanto, E. (2014). ‏Role of ‎pomegranate ‎and citrus fruit juices in colon cancer prevention. World J Gastroenterol; 20‎‎(16): 4618-4625.‎
Khorasani Esmaeili, A.; Mat Taha, R.; Mohajer, S.; Banisalam, B. (2015). Antioxidant Activity ‎and Total ‎Phenolic and Flavonoid Content of Various Solvent Extracts from In Vivo and in ‎Vitro Grown Trifolium ‎pratense L. (Red Clover). Biomed Res Int; 643285.‎
Kim, H.; Moon, J.Y.; Mosaddik, A.; Cho, S.K. (2010). Induction of apoptosis in human ‎cervical ‎carcinoma HeLa cells by polymethoxylated flavone-rich Citrus grandis Osbeck ‎‎(Dangyuja) leaf extract. ‎Food Chem Toxicol; 48(8-9): 2435-2442.‎
Kim, M.Y.; Bo, H.H.; Choi, E.O.; Kwon, D.H.; Kim, H.J.; Ahn, K.I.; Ji, S.Y.; Jeong, J.W.; ‎Park, S.H.; ‎Hong, S.H.; Kim, G.Y.; Park, C.; Kim, H.S.; Moon, S.K.; Yun, S.J.; Kim, W.J.; ‎Choi, Y.H. (2018). Induction of ‎Apoptosis by Citrus unshiu Peel in Human Breast Cancer MCF-7 ‎Cells: Involvement of ROS-Dependent ‎Activation of AMPK. Biol Pharm Bull; 41(5): 713-‎‎721.‎
Levings, D.C.; Wang, X.; Kohlhase, D.; ‏Bell, D.A.; Slattery, M. (2018). A distinct class of ‎antioxidant ‎response elements is consistently activated in tumors with NRF2 mutations. ‎Redox Biol; 19: 235-249.‎
Liang, S.; Lv, G.; Chen, W.; Jiang, J.; Wang, J. (2014). Citrus fruit intake and bladder cancer ‎risk: a ‎meta-analysis of observational studies. Int J Food Sci Nutr; 65(7): 893-898.‎
Marleau, A.M.; McDonald, G.; Koropatnick, J.; Chen, C.S.; Koos, D. (2012). Reduction ‎of ‎tumorigenicity by placental extracts. Anticancer Res; 32(4): 1153-1161.‎
Soong, Y.Y.; Tan, ‏S.P.; Leong, L.P.; Henry, J.K. (2014). Total antioxidant capacity and ‎starch ‎digestibility of muffins baked with rice, wheat, oat, corn and barley flour. Food Chem; ‎‎164: 462-469.‎
Wu, M.F.; Jian, Z.H.; Huang, J.Y.; Jan, C.F.; Nfor, O.N.; Jhang, K.M.; Ku, W.Y.; Ho, C.C.; Lung, C.‎C.; ‎Pan, H.H.; Wu, M.C.; Liaw; Y.P. (2016). Post-inhaled corticosteroid pulmonary tuberculosis and ‎pneumonia increases lung cancer in patients with COPD. BMC Cancer; 16 ‎‎(1): 778.‎
Yamasaki, M.; Hasegawa, S.; Takahashi, H.; ‏Kobayashi, Y.; Sakai, C.; Ashizawa, Y.; Asai, Y.; ‎Kanzaki, M.; Fukui, ‎T. (2015). Placental extracts induce the expression of antioxidant enzyme ‎genes and suppress melanogenesis in B16 melanoma cells. Nat Prod Res; 29(22): 2103-2106.‎
Yoon, S.W.; Jeong, J.S.; Kim, J.‏H.; Aggarwal, B.B. (2014). Cancer Prevention and ‎Therapy: ‎Integrating Traditional Korean Medicine Into Modern Cancer Care. Integr Cancer Ther; ‎‎13‎‎(4): 310-331.‎
Yu, X.; Sun, S.; Guo, Y.; Liu, Y.; Yang, D.; Li, G.; Lu, S. (2018). Citri Reticulatae Pericarpium (Chenpi): Botany, ethnopharmacology, phytochemistry, and pharmacology of a frequently ‎used traditional ‎Chinese medicine. J Ethnopharmacol; 220: 265-282.‎
Zhao, X.; Dai, X.; Wang, S.; Yang, T.; Yan, Y.; Zhu, G.; Feng, J.; Pan, B.; Sunagawa, M.; ‎Zhang, X.; Qian, Y.; Liu, ‎Y. (2018). Traditional Chinese Medicine Integrated with Chemotherapy ‎for Stage II-IIIA Patients with ‎Non-Small-Cell Lung Cancer after Radical Surgery: A ‎Retrospective Clinical Analysis with Small Sample ‎Size. Evid Based Complement Alternat Med. 4369027.