TY - JOUR
T1 - 2α-Hydroxyalantolactone from Pulicaria undulata
T2 - activity against multidrug-resistant tumor cells and modes of action
AU - Hegazy, Mohamed Elamir F.
AU - Dawood, Mona
AU - Mahmoud, Nuha
AU - Elbadawi, Mohamed
AU - Sugimoto, Yoshikazu
AU - Klauck, Sabine M.
AU - Mohamed, Nagla
AU - Efferth, Thomas
N1 - Funding Information:
Prof. Mohamed Hegazy gratefully acknowledges the financial support from Alexander von Humboldt Foundation “Georg Foster Research Fellowship for Experienced Researchers”. We are thankful to the Ph.D. stipend provided to MD by the Ministry of Higher Education and Scientific Research as well as Al-Neelain University, Khartoum, Sudan. We gratefully acknowledge the PhD stipend given by the German Academic exchange service (DAAD) to ME.
Funding Information:
Prof. Mohamed Hegazy gratefully acknowledges the financial support from Alexander von Humboldt Foundation ?Georg Foster Research Fellowship for Experienced Researchers?. We are thankful to the Ph.D. stipend provided to MD by the Ministry of Higher Education and Scientific Research as well as Al-Neelain University, Khartoum, Sudan. We gratefully acknowledge the PhD stipend given by the German Academic exchange service (DAAD) to ME. M-E.F. Hegazy and T. Efferth devised the project idea, designed and proof outlines. M-E.F. Hegazy performed all experiments. M. Dawood contributed with Western blotting and data analyses. N. Mahmoud contributed with quantitative real-time PCR (qRT-PCR) and data analyses. M. Elbadawi contributed with immunofluorescence staining. N. Mohamed collected the plant material and re-isolated the compound under study. Y. Sugimoto provided the HEK293/ABCB5 cells. S.M. Klauck run the microarray hybridizations. All authors drafted and contributed to manuscript preparation. All authors read the manuscript and approved the manuscript final version. T. Efferth supervised the project. All data were generated in-house, and no paper mill was used. All authors agree to be accountable for all aspects of work ensuring integrity and accuracy.
Publisher Copyright:
© 2020 Elsevier GmbH
PY - 2021/1
Y1 - 2021/1
N2 - Background: Sesquiterpene lactones having α-methylene-γ-lactone moiety are promising natural metabolites showing various biological activity. One of the major metabolites isolated from Pulicaria undulata, 2α-hydroxyalantolactone (PU-1), has not been investigated in detail yet. Multidrug resistance (MDR) represents a major obstacle for cancer chemotherapy and the capability of novel natural products to overcoming MDR is of great interest. Purpose: Exploring the molecular modes of action for potent natural product metabolites. Methods: The resazurin reduction assay was employed to evaluate the cytotoxicity of PU-1 on sensitive and their corresponding drug-resistant cell lines (overexpressing P-glycoprotein, BCRP, ABCB5, ΔEGFR, or TP53 knockout). Gene expression profiling was performed by transcriptome-wide mRNA microarray in the human CCRF-CEM leukemic cells after treatment with PU-1. The top significantly up- or down-regulated genes were identified by Chipster program and analyzed using Ingenuity Pathway Analysis (IPA) software. Finally, flow cytometry and Western blotting were performed for cell cycle analyses and apoptosis detection. Results: The sesquiterpene lactone, PU-1, showed potent cytotoxicity towards the drug-sensitive and -resistant cell lines. Transcriptome-wide mRNA expression profiling and pathway analysis pointed to genes involved in DNA damage response and G2/M cell cycle arrest. G2/M arrest was verified by flow cytometry and further confirmed by the upregulation of p21 and downregulation of p-CDC25C expression in Western blotting. Moreover, the suggested DNA damage checkpoint regulation was confirmed by immunofluorescence and Western blotting by upregulation of pS345 Chk1, p-H3 and γ-H2AX. Furthermore, PU-1 inhibited PI3K/AKT pathway, which is involved in signaling DNA damage and G2/M arrest. Cells ultimately induced apoptosis upon PU-1 treatment. Conclusions: PU-1 is a potent natural product inhibiting otherwise drug-resistant human tumor cell growth through DNA damage, G2/M cell cycle arrest and apoptosis.
AB - Background: Sesquiterpene lactones having α-methylene-γ-lactone moiety are promising natural metabolites showing various biological activity. One of the major metabolites isolated from Pulicaria undulata, 2α-hydroxyalantolactone (PU-1), has not been investigated in detail yet. Multidrug resistance (MDR) represents a major obstacle for cancer chemotherapy and the capability of novel natural products to overcoming MDR is of great interest. Purpose: Exploring the molecular modes of action for potent natural product metabolites. Methods: The resazurin reduction assay was employed to evaluate the cytotoxicity of PU-1 on sensitive and their corresponding drug-resistant cell lines (overexpressing P-glycoprotein, BCRP, ABCB5, ΔEGFR, or TP53 knockout). Gene expression profiling was performed by transcriptome-wide mRNA microarray in the human CCRF-CEM leukemic cells after treatment with PU-1. The top significantly up- or down-regulated genes were identified by Chipster program and analyzed using Ingenuity Pathway Analysis (IPA) software. Finally, flow cytometry and Western blotting were performed for cell cycle analyses and apoptosis detection. Results: The sesquiterpene lactone, PU-1, showed potent cytotoxicity towards the drug-sensitive and -resistant cell lines. Transcriptome-wide mRNA expression profiling and pathway analysis pointed to genes involved in DNA damage response and G2/M cell cycle arrest. G2/M arrest was verified by flow cytometry and further confirmed by the upregulation of p21 and downregulation of p-CDC25C expression in Western blotting. Moreover, the suggested DNA damage checkpoint regulation was confirmed by immunofluorescence and Western blotting by upregulation of pS345 Chk1, p-H3 and γ-H2AX. Furthermore, PU-1 inhibited PI3K/AKT pathway, which is involved in signaling DNA damage and G2/M arrest. Cells ultimately induced apoptosis upon PU-1 treatment. Conclusions: PU-1 is a potent natural product inhibiting otherwise drug-resistant human tumor cell growth through DNA damage, G2/M cell cycle arrest and apoptosis.
KW - 2α-Hydroxyalantolactone
KW - Cytotoxicity
KW - DNA damage
KW - G2/M cell cycle arrest
KW - Mode of action
KW - Pulicaria undulata
UR - http://www.scopus.com/inward/record.url?scp=85098928297&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85098928297&partnerID=8YFLogxK
U2 - 10.1016/j.phymed.2020.153409
DO - 10.1016/j.phymed.2020.153409
M3 - Article
C2 - 33341310
AN - SCOPUS:85098928297
SN - 0944-7113
VL - 81
JO - Phytomedicine
JF - Phytomedicine
M1 - 153409
ER -
