Synergistic Cytotoxicity of Renieramycin M and Doxorubicin in MCF-7 Breast Cancer Cells
Renieramycin M (RM) is a KCN-stabilized tetrahydroisoquinoline purified from the blue sponge <i>Xestospongia</i> sp., with nanomolar IC<sub>50</sub>s against several cancer cell lines. Our goal is to evaluate its combination effects with doxorubicin (DOX) in estrogen receptor...
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doaj-03a730071b2b4d469ce4178e708fed972020-11-25T02:04:18ZengMDPI AGMarine Drugs1660-33972019-09-0117953610.3390/md17090536md17090536Synergistic Cytotoxicity of Renieramycin M and Doxorubicin in MCF-7 Breast Cancer CellsJortan O. Tun0Lilibeth A. Salvador-Reyes1Michael C. Velarde2Naoki Saito3Khanit Suwanborirux4Gisela P. Concepcion5The Marine Science Institute, University of the Philippines Diliman, Quezon City 1101, PhilippinesThe Marine Science Institute, University of the Philippines Diliman, Quezon City 1101, PhilippinesInstitute of Biology, University of the Philippines Diliman, Quezon City 1101, PhilippinesGraduate School of Pharmaceutical Sciences, Meiji Pharmaceutical University, Tokyo 204-8588, JapanDepartment of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Center for Bioactive Natural Products from Marine Organisms and Endophytic Fungi (BNPME), Chulalongkorn University, Pathumwan, Bangkok 10330, ThailandThe Marine Science Institute, University of the Philippines Diliman, Quezon City 1101, PhilippinesRenieramycin M (RM) is a KCN-stabilized tetrahydroisoquinoline purified from the blue sponge <i>Xestospongia</i> sp., with nanomolar IC<sub>50</sub>s 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 IC<sub>95</sub> 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.https://www.mdpi.com/1660-3397/17/9/536renieramycin Mdoxorubicinsynergistic combination chemotherapyreal-time profilingcell cyclegene expression profilingapoptosisbreast cancerblue spongeDNA damage response |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jortan O. Tun Lilibeth A. Salvador-Reyes Michael C. Velarde Naoki Saito Khanit Suwanborirux Gisela P. Concepcion |
spellingShingle |
Jortan O. Tun Lilibeth A. Salvador-Reyes Michael C. Velarde Naoki Saito Khanit Suwanborirux Gisela P. Concepcion Synergistic Cytotoxicity of Renieramycin M and Doxorubicin in MCF-7 Breast Cancer Cells Marine Drugs renieramycin M doxorubicin synergistic combination chemotherapy real-time profiling cell cycle gene expression profiling apoptosis breast cancer blue sponge DNA damage response |
author_facet |
Jortan O. Tun Lilibeth A. Salvador-Reyes Michael C. Velarde Naoki Saito Khanit Suwanborirux Gisela P. Concepcion |
author_sort |
Jortan O. Tun |
title |
Synergistic Cytotoxicity of Renieramycin M and Doxorubicin in MCF-7 Breast Cancer Cells |
title_short |
Synergistic Cytotoxicity of Renieramycin M and Doxorubicin in MCF-7 Breast Cancer Cells |
title_full |
Synergistic Cytotoxicity of Renieramycin M and Doxorubicin in MCF-7 Breast Cancer Cells |
title_fullStr |
Synergistic Cytotoxicity of Renieramycin M and Doxorubicin in MCF-7 Breast Cancer Cells |
title_full_unstemmed |
Synergistic Cytotoxicity of Renieramycin M and Doxorubicin in MCF-7 Breast Cancer Cells |
title_sort |
synergistic cytotoxicity of renieramycin m and doxorubicin in mcf-7 breast cancer cells |
publisher |
MDPI AG |
series |
Marine Drugs |
issn |
1660-3397 |
publishDate |
2019-09-01 |
description |
Renieramycin M (RM) is a KCN-stabilized tetrahydroisoquinoline purified from the blue sponge <i>Xestospongia</i> sp., with nanomolar IC<sub>50</sub>s 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 IC<sub>95</sub> 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. |
topic |
renieramycin M doxorubicin synergistic combination chemotherapy real-time profiling cell cycle gene expression profiling apoptosis breast cancer blue sponge DNA damage response |
url |
https://www.mdpi.com/1660-3397/17/9/536 |
work_keys_str_mv |
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