Synergistic cytotoxicity of renieramycin M and doxorubicin in MCF-7 breast cancer cells
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PROVIDER: PRJNA512621 | ENA |
REPOSITORIES: ENA
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Project description:Renieramycin M (RM) is a KCN-stabilized tetrahydroisoquinoline purified from the blue sponge Xestospongia sp., with nanomolar IC50s against several cancer cell lines. Our goal is to evaluate its combination effects with doxorubicin (DOX) in estrogen receptor positive MCF-7 breast cancer cells. MCF-7 cells were treated simultaneously or sequentially with various combination ratios of RM and DOX for 72 h. Cell viability was determined using the MTT assay. Synergism or antagonism was determined using curve-shift analysis, Combination Index method and isobologram analysis. Synergism was observed with pharmacologically achievable concentrations of DOX when administered simultaneously, but not sequentially. The IC95 of RM and DOX after combination were reduced by up to 4-fold and 8-fold, respectively. To gain insights on the mechanism of synergy, real-time profiling, cell cycle analysis, apoptosis assays, and transcriptome analysis were conducted. The combination treatment displayed similar profile with DNA-damaging agents and induced a greater and faster cell killing. The combination treatment also showed an increase in apoptosis. DOX induced S and G2/M arrest while RM did not induce significant changes in the cell cycle. DNA replication and repair genes were commonly downregulated by RM and DOX. p53 signaling and cell cycle checkpoints were regulated by DOX while ErbB/PI3K-Akt, integrin and focal adhesion signaling were regulated by RM upon combination. Genes involved in cytochrome C release and interferon gamma signaling were regulated specifically in the combination treatment. This study serves as basis for in vivo studies and provides a rationale for using RM in combination with other anticancer drugs. We used microarray-based transcriptome profiling to gain insights to the mechanism of action of the individual drugs and the synergistic drug combination
2019-01-04 | GSE124597 | GEO
Project description:The combination of natural products with standard chemotherapeutic agents offers a promising strategy to enhance the efficacy of standard chemotherapy by reducing their dosage and side effects. The standard chemotherapeutic drug for breast and other cancer types- doxorubicin(DOX), also known as Adriamycin (an anthracycline) has several disadvantages including severe side effects and development of drug resistance. Recently, we reported the broad spectrum pharmacological activity and potential markers of Australian propolis extract (AP-1). In the present study, we explored the synergistic interactions of AP-1 with DOX against MCF-7 cells by implementing different synergy quantitation models. To identify the potential anticancer metabolites in AP-1, biochemometric and metabolomics-driven study was performed. Furthermore, we evaluated the molecular mechanisms involved by analysing the apoptotic profile (using flow cytometry and proteome array of 35 apoptotic proteins) and oxidative status (using reactive oxygen species detection) of the MCF-7 cells upon treatment with the most synergistic combination. The underlying synergistic mechanism against the MCF-7 cells was also studied using label-free quantification proteomics analysis. Five prenylated stilbenes were identified as the most discriminant metabolites in the most active AP-1 fraction, three of them previously isolated from AP-1 with IC50 values in the range 0.68-2.7 µM against the MCF-7 cells. Strong synergy was observed when AP-1 was combined with DOX in the ratio of 100:0.29 (w/w) as validated by different synergy quantitation models including combination index (CI), HSA, ZIP, LOEWE, and BLISS. AP-1 significantly enhanced the inhibitory effect of DOX (p < 0.05) against the MCF-7 cell proliferation in a dose-dependent manner with significant reduction of ROS production (p < 0.05; compared to DOX alone). AP-1 also enhanced the apoptotic effect of DOX significantly after 24 h of treatment with a shift of DOX-induced necrosis to apoptosis. Significant upregulation of catalase, HTRA2/Omi, FADD together with TRAIL-mediated apoptosis, DR5 and DR4 (p < 0.05) was observed which contributed to the anti-proliferative activity of AP-1 against the MCF-7 cells. The enhancement of pro-apoptotic p27, PON2 and catalase with reduction of anti-apoptotic XIAP, HSP60 and HIF-1α may be associated with the improved apoptosis in the MCF-7 cells observed in the Annexin V-7AAD flow cytometry analysis of the synergistic combination compared to the mono treatments. Furthermore, a significant increase of antioxidant proteins including catalase and PON2 was observed upon treatment with the synergistic combination which may be associated with the parallel enhancement of apoptosis. Shotgun proteomics in the synergistic combination-treated cells identified 21 significantly dysregulated proteins involving the TP53/ATM-regulated non-homologous end-joining pathway and double-strand breaks repairs, recruiting the overexpressed BRCA1 and curtailed RIF1 encoded proteins. The overexpression of UPF2 was noticed after the treatment with the synergistic combination which could assist to overcome doxorubicin resistance-associated long non-coding RNA and decline the metastasis of the MCF-7 cells. In summary, we propose a promising AP-1 and DOX synergistic combination against the MCF-7 cells and highlighted the possible synergistic mechanisms of action together with the key prenylated metabolites of AP-1. Further in vivo and clinical studies are warranted on this synergistic combination.
2021-07-26 | PXD026331 | Pride
Project description:Objective: To assess the role of aldoketoreductases and other doxorubicin pharmacokinetic or pharmacogenomic genes in doxorubicin cytotoxicity, resistance, DNA binding activity, and subcellular localization, Methods: We conducted a whole genome microarray study to identify differences in between doxorubicin-sensitive MCF-7cc cells and doxorubicin-resistant MCF-7Dox2-12 cells in terms of their expression of genes related to doxorubicin pharmacokinetics or pharmacodynamics. Targets were then validated by pharmacologic inhibition in conjunction with drug metabolite profiling, drug localization, drug cytotoxicity, and drug DNA binding studies. Results: 2063 differentially expressed transcripts were identified, including 17% and 43% of genes or gene families associated with doxorubicin pharmacokinetics or pharmacodynamics (p values of significance of 0.05 and <0.0001, respectively). The largest changes in the expression of genes associated with doxorubicin pharmacokinetics and pharmacodynamics were chiefly among the aldo-keto reductases (AKRs) Akr1c2, Akr1c3 and Akr1b10 which convert doxorubicin to doxorubicinol. We observed that doxorubicinol exhibits dramatically reduced drug toxicity, reduced drug DNA-binding activity, and altered drug subcellular localization to lysosomes. Pharmacologic inhibition of these AKRs in MCF-7Dox2-12 cells restored drug cytotoxicity, and drug localization to the nucleus. Conclusion: These findings demonstrate the utility of using curated pharmacokinetic and pharmacodynamic knowledgebases to identify highly relevant genes associated with doxorubicin resistance. The products of one or more of these genes could effectively be shown to alter the drug’s properties, while inhibiting them restored drug DNA binding, cytotoxicity, and subcellular localization. Doxorubicin resistant cell lines of breast MCF-7 cells were generated for gene expression profilling. Two colour microarray of Agilent whole human genome nucleotide arrays was conducted with four labelling replicates of both forward and reverse labellings plus another set of 8 arrays with forward labelling. Sixteen arrays were used for this experiments. The co-cultured control cells MCF-7cc12 was generated by parallel selection process in the absence of drug.
2012-11-22 | E-GEOD-27254 | biostudies-arrayexpress
Project description:MCF-7 cells treated with 10μM doxorubicin for 4h followed by subsequent withdrawl of the drug and cultured up to 48h.Doxurubicin-treated cells and control cells were collected at 0,12,24,36,and 48h. Meanwhile,MCF-7 cells continuously exposed to a low dosage of doxorubicin at 0.1μM for 96h. Doxorubicin-treated cells and control cells were collected at 0,24,48,72 and 96h.
2019-04-11 | ST001170 | MetabolomicsWorkbench
Project description:Objective: To assess the role of aldoketoreductases and other doxorubicin pharmacokinetic or pharmacogenomic genes in doxorubicin cytotoxicity, resistance, DNA binding activity, and subcellular localization, Methods: We conducted a whole genome microarray study to identify differences in between doxorubicin-sensitive MCF-7cc cells and doxorubicin-resistant MCF-7Dox2-12 cells in terms of their expression of genes related to doxorubicin pharmacokinetics or pharmacodynamics. Targets were then validated by pharmacologic inhibition in conjunction with drug metabolite profiling, drug localization, drug cytotoxicity, and drug DNA binding studies. Results: 2063 differentially expressed transcripts were identified, including 17% and 43% of genes or gene families associated with doxorubicin pharmacokinetics or pharmacodynamics (p values of significance of 0.05 and <0.0001, respectively). The largest changes in the expression of genes associated with doxorubicin pharmacokinetics and pharmacodynamics were chiefly among the aldo-keto reductases (AKRs) Akr1c2, Akr1c3 and Akr1b10 which convert doxorubicin to doxorubicinol. We observed that doxorubicinol exhibits dramatically reduced drug toxicity, reduced drug DNA-binding activity, and altered drug subcellular localization to lysosomes. Pharmacologic inhibition of these AKRs in MCF-7Dox2-12 cells restored drug cytotoxicity, and drug localization to the nucleus. Conclusion: These findings demonstrate the utility of using curated pharmacokinetic and pharmacodynamic knowledgebases to identify highly relevant genes associated with doxorubicin resistance. The products of one or more of these genes could effectively be shown to alter the drug’s properties, while inhibiting them restored drug DNA binding, cytotoxicity, and subcellular localization.
2012-11-22 | GSE27254 | GEO
Project description:Renieramycin M (RM) is a KCN-stabilized tetrahydroisoquinoline purified from the blue sponge Xestospongia sp., with nanomolar IC50s against several cancer cell lines. Our goal is to evaluate its combination effects with doxorubicin (DOX) in estrogen receptor positive MCF-7 breast cancer cells. MCF-7 cells were treated simultaneously or sequentially with various combination ratios of RM and DOX for 72 h. Cell viability was determined using the MTT assay. Synergism or antagonism was determined using curve-shift analysis, combination index method and isobologram analysis. Synergism was observed with pharmacologically achievable concentrations of DOX when administered simultaneously, but not sequentially. The IC95 values of RM and DOX after combination were reduced by up to four-fold and eight-fold, respectively. To gain insights on the mechanism of synergy, real-time profiling, cell cycle analysis, apoptosis assays, and transcriptome analysis were conducted. The combination treatment displayed a similar profile with DNA-damaging agents and induced a greater and faster cell killing. The combination treatment also showed an increase in apoptosis. DOX induced S and G2/M arrest while RM did not induce significant changes in the cell cycle. DNA replication and repair genes were downregulated commonly by RM and DOX. p53 signaling and cell cycle checkpoints were regulated by DOX while ErbB/PI3K-Akt, integrin and focal adhesion signaling were regulated by RM upon combination. Genes involved in cytochrome C release and interferon gamma signaling were regulated specifically in the combination treatment. This study serves as a basis for in vivo studies and provides a rationale for using RM in combination with other anticancer drugs.
| S-EPMC6780817 | biostudies-other
Project description:Despite improvements in therapeutic strategies for treating breast cancers, tumor relapse and chemoresistance remain major issues in patient outcomes. Indeed, cancer cells display a metabolic plasticity allowing a quick adaptation to tumoral microenvironment and to cellular stresses induced by chemotherapy. Recently, long non-coding RNA molecules (lncRNAs) have emerged as important regulators of cellular metabolic orientation. In the present study, we addressed the role of the long non-coding RNA molecule (lncRNA) SAMMSON on the metabolic reprogramming and chemoresistance of MCF-7 breast cancer cells resistant to doxorubicin (MCF-7dox). Our results showed an overexpression of SAMMSON in MCF-7dox compared to doxorubicin-sensitive cells (MCF-7). Silencing of SAMMSON expression by siRNA in MCF-7dox cells resulted in a metabolic rewiring with improvement of oxidative metabolism, decreased mitochondrial ROS production, increased mitochondrial replication, transcription and translation and an attenuation of chemoresistance. These results highlight the role of SAMMSON in the metabolic adaptations leading to the development of chemoresistance in breast cancer cells. Thus, targeting SAMMSON expression levels represents a promising therapeutic route to circumvent doxorubicin resistance in breast cancers.
2021-11-10 | GSE188404 | GEO
Project description:The breast cancer cell line MCF-7 was treated with doxorubicin. Protein expression differences between treated and non-treated cells were semiquantitatively determined using the precursor acquisition independent from ion count (PAcIFIC) mass spectrometric method. The acquired proteomic data sets were searched for regulated Reactome pathways and Gene Ontology annotation terms using a new algorithm (SetRank). Using this approach, we described significantly changed pathways (p-value of 0.05 or less) already known to be influenced by chemotherapeutic drugs, such as chromatin organization, DNA binding, embryo development, condensed chromosome, sequence-specific DNA binding, response to oxidative stress and response to toxin. Additionally, we found pathways such as central nervous system neuron differentiation, neuron projection membrane and SNAP receptor activity explaining side-effects of doxorubicin chemotherapy, characterized as ‘chemo brain’, on a molecular level.
2015-11-09 | PXD002998 | Pride
Project description:Silencing branched-chain ketoacid dehydrogenase kinase exacerbates doxorubicin induced cytotoxicity in triple-negative breast cancer cells
| PRJNA685647 | ENA
Project description:The FDA approved drug Doxorubicin provokes copious irreversible cardiotoxicity and even increases the risk of heart failure. Considering the multiple and interacted molecular pathways in cancer, there is a big possibility that tumors are simultaneously sensitive to different drugs. This makes achievable to study the combinations of drug, having the virtues of less toxicity, higher efficacy and potentially antagonizing drug resistance in cancer therapy. In the present study, we addressed the synergistic effects of ginsenoside Rh2 on doxorubicin-treated breast cancer bearing mice. We showed that Rh2 significantly enhanced the anti-cancer effects of doxorubicin and greatly attenuated the cardiotoxicity. Transcriptomic changes can clearly distinguish the chemotherapeutic groups and non-treated control groups. Transcriptomic analysis domestrated that Rh2 protection involved in multiple vital pathways including cellular stress, apoptosis and inflammation.
2021-10-28 | GSE186555 | GEO