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. 2017 Feb;20(2):166-171.
doi: 10.22038/ijbms.2017.8242.

Cytotoxic and apoptotic effects of different extracts of Artemisia biennis Willd. on K562 and HL-60 cell lines

Zahra Tayarani-Najaran  1 , Farideh-Sadat Makki  2 , Nafiseh-Sadat Alamolhodaei  2 , Mahdi Mojarrab  3 , Seyed Ahmad Emami  4
Affiliations

Cytotoxic and apoptotic effects of different extracts of Artemisia biennis Willd. on K562 and HL-60 cell lines

Zahra Tayarani-Najaran et al. Iran J Basic Med Sci. 2017 Feb.

Abstract

Objectives: Artemisia is a genus of herbs and small shrubs forms an important part of natural vegetation in Iran. It has been reported that several Artemisia species possess anti-proliferative effects. Considering the value of this genus in anti-cancer researches we have chosen Artemisia biennis for cytotoxic and mechanistic studies.

Materials and methods: In this study we have investigated the cytotoxic and apoptotic effects of petroleum ether, dichloromethane, ethyl acetate, ethanol, and ethanol: water (1:1 v/v) extracts of A. biennis Willd. on two cancer human cell lines (K562 and HL-60) and J774 as normal cells.

Results: CH2Cl2 extract was found to have the highest anti-proliferative effect on cancer cells. IC50 values obtained in AlamarBlue® assay for CH2Cl2 extract were 64.86 and 54.31 µg/ml on K562 and HL-60 cells respectively. In flow cytometry histogram of the cells treated with CH2Cl2 extract, sub-G1 peak was induced. DNA fragmentation, increased in the level of Bax and cleavage of PARP protein all showed the induction of apoptosis with CH2Cl2 extract after 48 hr contact with cells.

Conclusion: The results can corroborate the cytotoxic and apoptotic effects of the CH2Cl2 extract of A. biennis on the K562 and HL-60 cancer cell lines.

Keywords: Apoptosis; Artemisia biennis; Asteraceae; Leukemic cell lines.

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Figures

Figure 1
Figure 1
Extraction scheme of PE, CH2Cl2, EtOAc, EtOH and EtOH/H2O (1:1 v/v) extracts of Artemisia biennis
Figure 2
Figure 2
The dose-dependent effects of extracts on the growth of K562 and HL-60 cells and normal J774 cells. PE and CH2Cl2 extracts exhibited cytotoxic activity against apoptosis-proficient HL-60 and apoptosis-resistant K562 cells with minimal cytotoxic effects on normal J774 cells. Values were mean±SEM of at least three independent experiments, each in triplicates
Figure 3
Figure 3
PI staining and flow cytometry analysis of CH2Cl2 extract (0, 25, 50, 100 µg/ml) induced apoptosis in K562 and HL-60 cells
Figure 4
Figure 4
Proteolytic cleavage of poly (ADP-ribose) polymerase (PARP) and increasing in the level of Bax protein in HL-60 cells after 48 hr exposure to CH2Cl2 extracts (0, 25, 50, 100 µg/ml). Β-Actin was used as a loading control. All Western blots were representative of 3 independent experiments
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