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Evaluation of the cytotoxicity, cell-cycle arrest, and apoptotic induction by Euphorbia hirta in MCF-7 breast cancer cells.
Pharm Biol. 2016 Jul; 54(7):1223-36.PB

Abstract

CONTEXT

Euphorbia hirta L. (Euphorbiaceae) has been used as a folk remedy in Southeast Asia for the treatment of various ailments.

OBJECTIVE

The current study evaluates the cytotoxicity, cell-cycle arrest, and apoptotic induction by E. hirta in MCF-7 breast cancer cells.

MATERIALS AND METHODS

Cytotoxic activity of methanol extract of whole part of E. hirta was determined by the MTT assay at various concentrations ranging from 1.96 to 250.00 µg/mL in MCF-7 cells. Cell morphology was assessed by light and fluorescence microscopy. Apoptosis and cell-cycle distribution were determined by annexin V staining and flow cytometry. DNA fragmentation, caspase activity, and reactive oxygen species (ROS) assays were performed using the commercially available kits. To identify the cytotoxic fraction, E. hirta extract was subjected to bioassay-guided fractionation.

RESULTS

Euphorbia hirta exhibited significant inhibition of the survival of MCF-7 cells and the half inhibitory concentration (IC50) values was 25.26 µg/mL at 24 h. Microscopic studies showed that E. hirta-treated cells exhibited marked morphological features characteristic of apoptosis. Euphorbia hirta extract also had an ignorable influence on the LDH leakage and generating intracellular ROS. The flow cytometry study confirmed that E. hirta extract induced apoptosis in MCF-7 cells. Euphorbia hirta also resulted in DNA fragmentation in MCF-7 cells. Moreover, E. hirta treatment resulted in the accumulation of cells at the S and G2/M phases as well as apoptosis. The caspase activity study revealed that E. hirta extract induced apoptosis through the caspase-3-independent pathway by the activation of caspase-2, 6, 8, and 9. Euphorbia hirta hexane fraction, namely HFsub4 fraction, demonstrated highest activity among all the fractions tested with an IC50 value of 10.01 µg/mL at 24 h.

DISCUSSION AND CONCLUSION

This study revealed that E. hirta induced apoptotic cell death and suggests that E. hirta could be used as an apoptosis-inducing anticancer agent for breast cancer treatment with further detailed studies.

Authors+Show Affiliations

a Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, USM , Penang , Malaysia .b RIKEN-USM Joint Research Unit , RIKEN , Wako , Saitama , Japan .c School of Biological Sciences, Universiti Sains Malaysia , Penang , Malaysia .d Advanced Medical and Dental Institute, Universiti Sains Malaysia , Penang , Malaysia .a Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, USM , Penang , Malaysia .e Faculty of Dentistry , Dental Research & Training Unit, and Oral Cancer Research and Coordinating Centre (OCRCC), University of Malaya , Kuala Lumpur , Malaysia , and.a Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, USM , Penang , Malaysia .f Faculty of Health, Nanomedicine - Laboratory of Immunology and Molecular Biomedical Research (LIMBR) , School of Medicine (SoM), Institute for Frontier Materials (IFM), Deakin University , Waurn Ponds , Victoria , Australia.a Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, USM , Penang , Malaysia .

Pub Type(s)

Journal Article

Language

eng

PubMed ID

26154521

Citation

Kwan, Yuet Ping, et al. "Evaluation of the Cytotoxicity, Cell-cycle Arrest, and Apoptotic Induction By Euphorbia Hirta in MCF-7 Breast Cancer Cells." Pharmaceutical Biology, vol. 54, no. 7, 2016, pp. 1223-36.
Kwan YP, Saito T, Ibrahim D, et al. Evaluation of the cytotoxicity, cell-cycle arrest, and apoptotic induction by Euphorbia hirta in MCF-7 breast cancer cells. Pharm Biol. 2016;54(7):1223-36.
Kwan, Y. P., Saito, T., Ibrahim, D., Al-Hassan, F. M., Ein Oon, C., Chen, Y., Jothy, S. L., Kanwar, J. R., & Sasidharan, S. (2016). Evaluation of the cytotoxicity, cell-cycle arrest, and apoptotic induction by Euphorbia hirta in MCF-7 breast cancer cells. Pharmaceutical Biology, 54(7), 1223-36. https://doi.org/10.3109/13880209.2015.1064451
Kwan YP, et al. Evaluation of the Cytotoxicity, Cell-cycle Arrest, and Apoptotic Induction By Euphorbia Hirta in MCF-7 Breast Cancer Cells. Pharm Biol. 2016;54(7):1223-36. PubMed PMID: 26154521.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Evaluation of the cytotoxicity, cell-cycle arrest, and apoptotic induction by Euphorbia hirta in MCF-7 breast cancer cells. AU - Kwan,Yuet Ping, AU - Saito,Tamio, AU - Ibrahim,Darah, AU - Al-Hassan,Faisal Muti Saleh, AU - Ein Oon,Chern, AU - Chen,Yeng, AU - Jothy,Subramanion L, AU - Kanwar,Jagat R, AU - Sasidharan,Sreenivasan, Y1 - 2015/07/08/ PY - 2015/7/9/entrez PY - 2015/7/15/pubmed PY - 2017/2/9/medline KW - Anticancer KW - G2/M phase arrest KW - S phase arrest KW - caspase SP - 1223 EP - 36 JF - Pharmaceutical biology JO - Pharm Biol VL - 54 IS - 7 N2 - CONTEXT: Euphorbia hirta L. (Euphorbiaceae) has been used as a folk remedy in Southeast Asia for the treatment of various ailments. OBJECTIVE: The current study evaluates the cytotoxicity, cell-cycle arrest, and apoptotic induction by E. hirta in MCF-7 breast cancer cells. MATERIALS AND METHODS: Cytotoxic activity of methanol extract of whole part of E. hirta was determined by the MTT assay at various concentrations ranging from 1.96 to 250.00 µg/mL in MCF-7 cells. Cell morphology was assessed by light and fluorescence microscopy. Apoptosis and cell-cycle distribution were determined by annexin V staining and flow cytometry. DNA fragmentation, caspase activity, and reactive oxygen species (ROS) assays were performed using the commercially available kits. To identify the cytotoxic fraction, E. hirta extract was subjected to bioassay-guided fractionation. RESULTS: Euphorbia hirta exhibited significant inhibition of the survival of MCF-7 cells and the half inhibitory concentration (IC50) values was 25.26 µg/mL at 24 h. Microscopic studies showed that E. hirta-treated cells exhibited marked morphological features characteristic of apoptosis. Euphorbia hirta extract also had an ignorable influence on the LDH leakage and generating intracellular ROS. The flow cytometry study confirmed that E. hirta extract induced apoptosis in MCF-7 cells. Euphorbia hirta also resulted in DNA fragmentation in MCF-7 cells. Moreover, E. hirta treatment resulted in the accumulation of cells at the S and G2/M phases as well as apoptosis. The caspase activity study revealed that E. hirta extract induced apoptosis through the caspase-3-independent pathway by the activation of caspase-2, 6, 8, and 9. Euphorbia hirta hexane fraction, namely HFsub4 fraction, demonstrated highest activity among all the fractions tested with an IC50 value of 10.01 µg/mL at 24 h. DISCUSSION AND CONCLUSION: This study revealed that E. hirta induced apoptotic cell death and suggests that E. hirta could be used as an apoptosis-inducing anticancer agent for breast cancer treatment with further detailed studies. SN - 1744-5116 UR - https://www.unboundmedicine.com/medline/citation/26154521/Evaluation_of_the_cytotoxicity_cell_cycle_arrest_and_apoptotic_induction_by_Euphorbia_hirta_in_MCF_7_breast_cancer_cells_ L2 - https://www.tandfonline.com/doi/full/10.3109/13880209.2015.1064451 DB - PRIME DP - Unbound Medicine ER -