Cytotoxic and apoptotic effects of root extract and tanshinones isolated from Perovskia abrotanoides Kar

Document Type : Original Article

Authors

1 Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran

2 Department of Pharmacognosy, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

3 Medical Toxicology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

4 Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

5 Medical Toxicology Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

6 Pharmacological Research Center of Medicinal Plants, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

7 Department of Pharmacology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Objective(s):Perovskia abrotanoides Kar., from family Lamiaceae, is a little known medicinal plant growing in various regions of Iran. In the present study, cryptotanshinone (CT), tanshinone 2A (Tan2A), and hydroxycryptotanshinone (HCT) were isolated and purified from the roots of P. abrotanoides. In addition, cytotoxic and apoptotic effects of total root extract (TE) and three purified tanshinones were investigated in human cervical carcinoma (HeLa) and human breast cancer (MCF-7) cell lines. 
Materials and Methods: Alamar Blue® assay was used to determine cell viability. Cell apoptotic rate was detected using propidium iodide staining of DNA fragmentation by flow cytometry (sub-G1 peak). The PARP cleavage, as the sign of apoptosis, was investigated by Western blotting.
Results: The results revealed that CT, Tan2A, HCT, and TE exhibited significant cytotoxicity in cancer cell lines. All of these compounds caused apoptosis in treated cells and induced sub-G1 peak in the related flow cytometry histograms. HCT was found to have the highest anti-proliferative activity on cancer cells. Western blotting analysis showed cleavage of PARP protein in MCF-7 cells treated with purified tanshinones and TE after 48 hr contact with cells.
Conclusion: The findings suggest that root extract of P. abrotanoides and purified tanshinones especially Tan2A and HCT have cytotoxic and apoptotic effects against cancer cell lines. So, they may serve as potential cytotoxic agents for future investigations.

Keywords


1.Park EJ, Zhao YZ, Kim YC, Sohn DH. PF2401-SF standardized fraction of Salvia miltiorrhiza and its constituents, tanshinone I, tanshinone IIA, and cryptotanshinone, protect primary cultured rat hepatocytes from bile acid-induced apoptosis by inhibiting JNK phosphorylation. Food Chem Toxicol 2007; 45: 1891-1898.
2.Cragg GM, Newman DJ. Natural products drug discovery in the next millennium. Pharm Biol 2001; 39: 8-17.
3.Sairafianpour M, Christensen J, Staerk D, Budnik BA, Kharazmi A, Bagherzadeh K et al. Leishmanicidal, antiplasmodial, and cytotoxic activity of novel diterpenoid 1,2-quinones from Perovskia abrotanoides: New source of tanshinones. J Nat Prod 2001; 64: 1398-1403.
4.Hosseinzadeh H, Amel S. Antinociceptive effects of the aerial parts of Perovskia abrotanoides extracts in mice. Iran Red Crescent Med J 2001; 4: 15-17.
5.Zaker A, Sykora C, Gossnitzer F, Abrishamchi P, Asili J, Mousavi SH et al. Effects of some elicitors on tanshinone production in adventitious root cultures of Perovskia abrotanoides Karel. Ind Crops Prod 2015; 67: 97-102.
6.Kim EJ, Jung SN, Son KH, Kim SR, Ha TY, Park MG et al. Antidiabetes and antiobesity effect of cryptotanshinone via activation of AMP-activated protein kinase. Mol Pharmacol 2007; 72: 62–72.
7.Bai A, Lu N, Gou Y, Fan X. Tanshinone IIA ameliorates trinitrobenzene sulfonic acid (TNBS)-induced murine colitis. Digest Dis Sci 2008; 53: 421–428.
8.Park EJ, Zhao YZ, Kim YC, Sohn DH. Preventive effects of a purified extract isolated from Salvia miltiorrhiza enriched with tanshinone I, tanshinone IIA and cryptotanshinone on hepatocyte injury in vitro and in vivo. Food Chem Toxicol 2009; 47: 2742-2748.
9.Cao EH, Liu XQ, Wang JJ, Xu NF. Effect of natural antioxidant tanshinone II-A on DNA damage by lipid peroxidation in liver cells. Free Radic Bio Med 1996; 20: 801-806.
10.Pan TL, Hung YC, Wang PW, Chen ST, Hsu TK, Sintupisut N et al. Functional proteomic and structural insights into molecular targets related to the growth inhibitory effect of tanshinone IIA on HeLa cells. Proteomics 2010; 10: 914-929.
11.Liu JJ, Zhang Y, Lin DJ, Zhiao RZ. Tanshinone IIA inhibits leukemia THP-1 cell growth by induction of apoptosis. Oncol Rep 2009; 21: 1075-1081.
12.Lee DS, Lee SH, Noh JG, Hong SD. Antibacterial activities of cryptotanshinone and dihydrotanshinone I from medicinal herb, Salvia miltiorrhiza Bunge. Biosci Biotechnol Biochem 1999; 63: 2236-2239.
13.Fan GW, Gau XM, Wang H, Zhu Y, Zhang J, Hu LM et al. The anti-inflammatory activities of tanshinone IIA, an active component of TCM, are mediated by estrogen receptor activation and inhibition of iNOS. J Steroid Biochem Mol Biol 2009; 113: 275–280.
14.Su CC, Lin YH. Tanshinone IIA inhibits human breast cancer cells through increased Bax to Bcl-xL ratios. Int J Mol Med 2008; 22: 357-361.
15.Wang X, Wei Y, Yuan S, Liu G, Lu Y, Zhang J et al. Potential anticancer activity of tanshinone IIA against human breast cancer. Int J Cancer 2005; 116: 799-807.
16.Gong Y, Li Y, Lu Y, Li L, Abdolmaleky H, Blackburn GL et al. Bioactive tanshinones in Salvia miltiorrhiza inhibit the growth of prostate cancer cells in vitro and in mice. Int J Cancer 2011; 129: 1042-1052.
17.Park IJ, Kim MJ, Park OJ, Park MG, Choe W, Kang I et al. Cryptotanshinone sensitizes DU145 prostate cancer cells to Fas (APO1/CD95)-mediated apoptosis through Bcl-2 and MAPK regulation. Cancer Lett 2010; 298: 88-98.
18.Jiao JW, Wen F. Tanshinone IIA acts via p38 MAPK to induce apoptosis and the down-regulation of ERCC1 and lung-resistance protein in cisplatin-resistant ovarian cancer cells. Onco Rep 2010; 25: 781-788.
19.Lee WY, Chiu LC, Yeung JH. Cytotoxicity of major tanshinones isolated from Danshen (Salvia miltiorrhiza) on HepG2 cells in relation to glutathione perturbation. Food Chem Toxicol 2008; 46: 328–338.
20.Tavakoli J, Miar S, Majid Zadehzare M, Akbari H. Evaluation of effectiveness of herbal Medication in cancer care: A review study. Iran J Cancer Prev 2012; 5: 144-156.
21.Tayarani-Najaran Z, Amiri A, Karimi G, Emami SA, Asili J, Mousavi SH. Comparative studies of cytotoxic and apoptotic properties of different extracts and the essential oil of Lavandula angustifolia on malignant and normal cells. Nutr Cancer 2014; 66: 424-434.
22.Liu JJ, Liu WD, Yang HZ, Zhang Y, Fang ZG, Liu PQ et al. Inactivation of PI3k/Akt signaling pathway and activation of caspase-3 are involved in tanshinone I-induced apoptosis in myeloid leukemia cells in vitro. Ann Hematol 2010; 89: 1089-1097.
23.Prajitha V, Thoppil JE. Cytotoxic and apoptotic activities of extract of Amaranthus spinosus L. in Allium cepa and human erythrocytes. Cytotechnology 2017; 69:123–133.
24.Zhou L, Chan WK, Xu N, Xiao K, Luo H, Luo KQ et al. Tanshinone IIA, an isolated compound from Salvia miltiorrhiza Bunge, induces apoptosis in HeLa cells through mitotic arrest. Life Sci 2008; 83: 394-403.
25.Nizamutdinova IT, Lee GW, Lee JS, Cho MK, Son KH, Jeon SJ et al. Tanshinone I suppresses growth and invasion of human breast cancer cells, MDA-MB-231, through regulation of adhesion molecules. Carcinogenesis 2008; 29: 1885-1892.
26.Kasaian J, Iranshahi M, Masullo M, Piacente S, Ebrahimi F, Iranshahi M. Sesquiterpene lactones from Ferula oopoda and their cytotoxic properties. J Asian Nat Prod Res 2014; 16: 248–253.
27.Tayarani-Najaran Z, Mousavi SH, Asili J, Emami SA. Growth-inhibitory effect of Scutellaria lindbergii in human cancer cell lines. Food Chem Toxicol 2010; 48: 559-604.
28.Ashwini S, Ezhilarasan D, Anitha R. Cytotoxic effect of Caralluma fimbriata against human colon cancer cells. Pharmacogn J 2017; 9: 204-207.
29.Rashid M, Seghatoleslam A, Namavari M, Amiri A, Fahmidehkar MA, Ramezani A et al. Selective Cytotoxicity and apoptosis-induction of Cyrtopodion scabrum extract against digestive cancer cell lines. Int J Cancer Manag 2017; 10:e8633.
30.Sahranavard S, Naghibi F, Mosaddegh M, Esmaeili S, Sarkhail P, Taghvaei M et al. Cytotoxic activities of selected medicinal plants from Iran and phytochemical evaluation of the most potent extract. Res Pharm Sci 2009; 4: 133-137.
31.Liu J, Shen HM, Ong CN. Salvia miltirrhiza inhibits cell growth and induces apoptosis in human hepatoma HepG2 cells. Cancer Lett 2000; 153: 85-93.
32.Mosaddik MA. In vitro cytotoxicity of tanshinones isolated from Salvia miltiorrhiza Bunge against P388 lymphocytic leukemia cells. Phytomedicine 2003; 10: 682-685.
33.Tayarani-Najaran Z, Makki FS, Alamolhodaei NS, Mojarrab M, Emami SA. Cytotoxic and apoptotic effects of different extracts of Artemisia biennis Willd. on K562 and HL-60 cell lines. Iran J Basic Med Sci 2017; 20: 166-171.
34.Mothana RA, Jansen R, Gruenert R, Bednarski PJ, Lindequist U. Antimicrobial and cytotoxic abietane diterpenoids from the roots of Meriandera benghalensis (Roxb.) Benth. Pharmazie 2009; 64: 613-615.
35.Nordin ML, Abdul Kadir A, Zakaria ZA, Othman F, Abdullah R, Abdullah MN.  Cytotoxicity and apoptosis induction of Ardisia crispa and its solvent partitions against Mus musculus mammary carcinoma cell line (4T1). Evid Based Complement Alternat Med 2017; 2017:9368079.
36.Taraphdar AK, Roy M, Bhattacharya RK. Natural products as inducers of apoptosis: implication for cancer therapy and prevention. Curr Sci 2001; 80: 1387-1396.
37.Tayarani-Najaran Z, Emami SA, Asili J, Mirzaei A, Mousavi SH. Analyzing cytotoxic and apoptogenic properties of Scutellaria litwinowii root extract on cancer cell lines. Evid Based Complement Alternat Med 2011; 2011:160682.
38.Yoon Y, Kim YO, Jeon WK, Park HJ, Sung HJ. Tanshinone IIA isolated from Salvia miltiorrhiza Bunge induced apoptosis in HL60 human premyelocytic leukemia cell line. J Ethnopharmacol 1999; 68: 121-127.