Apoptosis-induction in human acute promyelocytic leukemia nb4 cells by bioactive compounds from redroot pigweed
Hamideh Bakhshayeshan-agdam,
1,* Nastaran sedghi ,
2 Majid mahdavi,
3 Seyed yahya salehi-lisar ,
4 Rouhollah motafakkerazad ,
5 Jafar razeghi ,
6
1. Department of Plant Sciences, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran.
2. Department of Animal Sciences, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran.
3. Department of Animal Sciences, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran.
4. Department of Plant Sciences, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran.
5. Department of Plant Sciences, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran.
6. Department of Plant Sciences, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran.
Abstract
Introduction
Leukemia cancer is currently one of the major causes of cancer-related deaths among humans. extracts of plants including weeds may be a potential source for treatments of leukemia cancer. redroot pigweed is weed species with well-known potential in bioactive compounds production. this plant has diverse therapeutic effects such as anti-cancer properties due to specific biochemicals which involving in its allelopathic interactions as well. we have proved anti-cancer potential of redroot pigweed on human acute promyelocytic leukemia (apl) cancer cell lines (nb4 cells) previously and amaranth ethanolic extract ic50 (160µg/ml) was determined by mtt assay. in this regard, to further investigation anti-cancer effects of redroot pigweed, the induction of apoptosis by amaranth ethanolic extract was studied and thereafter the existing compounds in the extract were identified.
Methods
In order to, ethanolic extract of redroot pigweed and annexin v-pi double staining assays were used for cell treatment and apoptotic cells investigation, respectively. subsequently, identification of amaranth phytocompounds via gc-ms analysis was conducted. quantitative assessment of apoptosis is evaluable by redistribution of the plasma membrane of cells (as hallmark of apoptosis) to visible phosphatidyl serine (ps) after double staining by annexin v-pi. after treatment of the cells with 160μg/ml amaranth extract for 24-72 h, the cells were investigated for the detection of early and late apoptosis and necrosis cells by flow cytometry.
Results
The translocation of phosphatidyl serine (annexin v positive) was increased after 24, 48 and 72 h. the rate of early apoptosis (annx+/pi−) and late apoptosis (annx+/pi+) and necrosis (annx−/pi+) in the cells treated with redroot pigweed ethanolic extract for 24, 48 and 72 h was calculated 25.70, 18.20 and 20.80 percent, 11.50, 25.20, 33.40 percent and 2.27, 3.60, 2.96 percent, respectively. these findings represent an increase in rate of apoptosis in a time-dependent manner. based on the gc-ms analysis results, 87 compounds were characterized and identified from the found peaks in the redroot pigweed extract in comparison to the mass spectra of the compounds with the nist library. briefly, gc-ms analysis showed that the ethanolic extract of redroot pigweed largely contained terpenoid compounds (51.71%) as the main bioactive compound groups. among terpenoid compounds, carvacrol (11.33%) was the key compound, followed by thymol (8.55%), alpha-pinene (4.73%), germacrene-d (4.61%), zingiberene (3.92%), and so on.
Conclusion
According to the obtained results, redroot pigweed contains various bioactive compounds with allelopathic and therapeutic properties and is recommended as a plant whit pharmaceutical importance. accordingly, screening novel herbs such plants existing in amaranthus genus with allelopathic traits and evaluation of possible potential therapeutic compounds can lead to improving traditional herbal medicine.
Keywords
Acute promyelocytic leukemia (apl), apoptosis, gc-ms analysis, redroot pigweed, ethanolic extract
