CYTOTOXIC AND APOPTOGENIC PROPERTIES OF
DRACOCEPHALUM KOTSCHYI AERIAL PART DIFFERENT
FRACTIONS ON CALU-6 AND MEHR-80 LUNG CANCER CELL LINES
TOOBA AHMADZADEH SANI1#, ELHAM MOHAMMADPOUR1,2#, AMENEH MOHAMMADI1#, TOKTAM MEMARIANI2#, MEHRAN VATANCHIAN YAZDI4#, RAMIN REZAEE5#, DANIELA CALINA6#, ANCA OANA DOCEA7#, MARINA GOUMENOU8,9#, LEILA ETEMAD10#, SHABNAM SHAHSAVAND11#*
1Research Center of Natural Products and Medicinal Plants, North Khorasan University of Medical Sciences, Bojnurd, Iran
2Department of Biochemistry, Payame Noor University, Mashhad, Iran
3Central Research Laboratory, North Khorasan University of Medical Sciences, Bojnurd, Iran
4Department of Anatomy, School of Medicine, North Khorasan University of Medical Sciences, Bojnurd, Iran
5Department of Physiology and Pharmacology, School of Medicine, North Khrosan University of Medical Sciences, Bojnurd, Iran
6Department of Clinical Pharmacology, University of Medicine and Pharmacy, Faculty of Pharmacy, Craiova, 200349, Romania
7Department of Toxicology, University of Medicine and Pharmacy, Faculty of Pharmacy, Craiova, 200349, Romania
8Laboratory of Toxicology, Medical School, University of Crete, Heraklion, Greece
9Pesticides Department, European Food Safety Authority, Parma, Italy
10Pharmaceutical Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
11Department of Physiology and Pharmacology, School of Medicine, North Khorasan University of Medical Sciences, P.O.
Box 94176-94735, Bojnurd, Iran
*corresponding author: shabnamshahsavand@gmail.com
#All authors contributed equally in preparing this manuscript and thus share first authorship
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Abstract:
Dracocephalum kotschyi is one of the Iranian medicinal plants. The aim of this study was to assess in vitro, the cytotoxic,
antiproliferative and apoptotic effects of this plant against lung cancer cell lines (Calu-6 and Mehr-80). Cell viability was
quantified by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay following 24 and 48 h exposure to
(12.5 􀀀 200 μg/mL) of plant methanolic extract, dichloromethane (CH2Cl2), ethyl acetate (EtOAc), hexane, water fractions
and the essential oil (EO). Further fractionation of primary total methanolic extract revealed luteolin (F10) as the major
component. The chemical constituents of EO were identified using gas chromatography linked mass spectrometry (GC MS)
analysis. The percentage of apoptotic cells was determined using propidium iodide (PI) staining of DNA fragments by flow
cytometry (sub G1 peak). Morphological changes in cells were identified. The most effective fractions were CH2Cl2 fraction,
EO and luteolin (F10) (IC50; 79.1 ± 5.36, 62.2 ± 6.94 and 56.32 ± 6.58 μg/mL in Calu-6 cells and 124.2 ± 5.78, 88.13 ± 4.29
and 78.32 ± 6.59 μg/mL in Mehr-80 cell line after 48 h respectively). The DNA fragmentation assay and morphological
microscopy also confirmed these data. Trans-citral (geranial) (15.65%), eucalyptol (9.32%), limonene (7.95%), beta-linalool
(4.64%), neryl-acetate (3.67%) and myrcene (3.36%) were identified as the major constituents of the EO. Dracocephalum
kotschyi different fractions and compounds have shown significant cytotoxic activities against Calu 6 and Mehr-80 cells.






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