TY - JOUR
T1 - Putative molecular determinants mediating sensitivity or resistance towards carnosic acid tumor cell responses
AU - Mahmoud, Nuha
AU - Saeed, Mohamed E.M.
AU - Sugimoto, Yoshikazu
AU - Klinger, Anette
AU - Fleischer, Edmond
AU - Efferth, Thomas
N1 - Funding Information:
The role of ABC transporters for MDR has been supported by numerous studies since the discovery of the first member of ABC transporter, P-glycoprotein (Gillet et al., 2007; Volm and Efferth, 2015b). Among the 49 identified human ABC transporters, P-glycoprotein (encoded by ABCB1/MDR1) and breast cancer resistance protein (encoded by ABCG2) have been linked with MDR of tumors in the clinics (Amiri-Kordestani et al., 2012; Natarajan et al., 2015; Shaffer et al., 2012). ABCB5 is a novel ABC transporter represents a marker for stem-like cancer cells, promoting progression and drug resistance (Luo et al., 2012). Despite the amount of studies investigated other ABC transporters-mediated chemoresistance in cell line models, their involvement in clinical drug resistance has not yet proven (Robey et al., 2018b). Continuous efforts were made to find or design potent and selective ABC transporter modulators, yet the third generation of P-glycoprotein inhibitors has failed in clinical trials (Eichhorn and Efferth, 2012; Zeino et al., 2014). Such compounds were relatively toxic and showed drug-drug interactions, if combined with conventional chemotherapy (Robey et al., 2018a). Recently, concepts emerged to develop drugs derived from natural origin to bypass or reverse MDR (Karthikeyan and Laxmanappa Hoti, 2015) (Wu et al., 2008). Interestingly, our findings revealed that cell lines overexpressing P-glycoprotein, ABCB5 or BCRP are similar or convergent in their response pattern to CA compared to the corresponding parental sensitive cells. This indicates that CA bypasses MDR mediated by the above three examined transporters. Other investigations explored the role of CA as inhibitor of P-glycoprotein's efflux activity. (Nabekura et al., 2010) found that CA increased daunorubicin and rhodamine 123 accumulation and re-sensitized human KB-C2 cervical carcinoma cells to vinblastine. In the context of food-drug interaction, CA increases the bioavailability of some P-glycoprotein anticancer drug substrates (Deferme and Augustijns, 2003).
Publisher Copyright:
© 2020 Elsevier GmbH
PY - 2020/10
Y1 - 2020/10
N2 - Background: Carnosic acid (CA) is one of the main constituents in rosemary extract. It possesses valuable pharmacological properties, including anti-oxidant, anti-inflammatory, anti-microbial and anti-cancer activities. Numerous in vitro and in vivo studies investigated the anticancer profile of CA and emphasized its potentiality for cancer treatment. Nevertheless, the role of multidrug-resistance (MDR) related mechanisms for CA's anticancer effect is not yet known. Purpose: We investigated the cytotoxicity of CA against known mechanisms of anticancer drug resistance (P-gp, ABCB5, BCRP, EGFR and p53) and determined novel putative molecular factors associated with cellular response towards CA. Study design: Cytotoxicity assays, bioinformatic analysis, flow cytometry and western blotting were performed to identify the mode of action of CA towards cancer cells. Methods: The cytotoxicity to CA was assessed using the resazurin assays in cell lines expressing the mentioned resistance mechanisms. A pharmacogenomic characterization of the NCI 60 cell line panel was applied via COMPARE, hierarchical cluster and network analyses. Flow cytometry was used to detect cellular mode of death and ROS generation. Changes in proteins-related to apoptosis were determined by Western blotting. Results: Cell lines expressing ABC transporters (P-gp, BCRP or ABCB5), mutant EGFR or p53 were not cross-resistant to CA compared to their parental counterparts. By pharmacogenomic approaches, we identified genes that belong to different functional groups (e.g. signal transduction, regulation of cytoskeleton and developmental regulatory system). These genes were predicted as molecular determinants that mediate CA tumor cellular responses. The top affected biofunctions included cellular development, cellular proliferation and cellular death and survival. The effect of CA-mediated apoptosis in leukemia cells, which were recognized as the most sensitive tumor type, was confirmed via flow cytometry and western blot analysis. Conclusion: CA may provide a novel treatment option to target refractory tumors and to effectively cooperate with established chemotherapy. Using pharmacogenomic approaches and network pharmacology, the relationship between cancer complexity and multi-target potentials of CA was analyzed and many putative molecular determinants were identified. They could serve as novel targets for CA and further studies are needed to translate the possible implications to clinical cancer treatment.
AB - Background: Carnosic acid (CA) is one of the main constituents in rosemary extract. It possesses valuable pharmacological properties, including anti-oxidant, anti-inflammatory, anti-microbial and anti-cancer activities. Numerous in vitro and in vivo studies investigated the anticancer profile of CA and emphasized its potentiality for cancer treatment. Nevertheless, the role of multidrug-resistance (MDR) related mechanisms for CA's anticancer effect is not yet known. Purpose: We investigated the cytotoxicity of CA against known mechanisms of anticancer drug resistance (P-gp, ABCB5, BCRP, EGFR and p53) and determined novel putative molecular factors associated with cellular response towards CA. Study design: Cytotoxicity assays, bioinformatic analysis, flow cytometry and western blotting were performed to identify the mode of action of CA towards cancer cells. Methods: The cytotoxicity to CA was assessed using the resazurin assays in cell lines expressing the mentioned resistance mechanisms. A pharmacogenomic characterization of the NCI 60 cell line panel was applied via COMPARE, hierarchical cluster and network analyses. Flow cytometry was used to detect cellular mode of death and ROS generation. Changes in proteins-related to apoptosis were determined by Western blotting. Results: Cell lines expressing ABC transporters (P-gp, BCRP or ABCB5), mutant EGFR or p53 were not cross-resistant to CA compared to their parental counterparts. By pharmacogenomic approaches, we identified genes that belong to different functional groups (e.g. signal transduction, regulation of cytoskeleton and developmental regulatory system). These genes were predicted as molecular determinants that mediate CA tumor cellular responses. The top affected biofunctions included cellular development, cellular proliferation and cellular death and survival. The effect of CA-mediated apoptosis in leukemia cells, which were recognized as the most sensitive tumor type, was confirmed via flow cytometry and western blot analysis. Conclusion: CA may provide a novel treatment option to target refractory tumors and to effectively cooperate with established chemotherapy. Using pharmacogenomic approaches and network pharmacology, the relationship between cancer complexity and multi-target potentials of CA was analyzed and many putative molecular determinants were identified. They could serve as novel targets for CA and further studies are needed to translate the possible implications to clinical cancer treatment.
KW - Chemotherapy
KW - Fak
KW - Lamiaceae
KW - Multidrug resistance
KW - Network pharmacology
KW - Phytochemicals
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UR - http://www.scopus.com/inward/citedby.url?scp=85087713248&partnerID=8YFLogxK
U2 - 10.1016/j.phymed.2020.153271
DO - 10.1016/j.phymed.2020.153271
M3 - Article
C2 - 32659679
AN - SCOPUS:85087713248
VL - 77
JO - Phytomedicine
JF - Phytomedicine
SN - 0944-7113
M1 - 153271
ER -
