Mithramycin A sensitizes therapy-resistant breast cancer stem cells toward genotoxic drug doxorubicin

• Published in: Translational research : the journal of laboratory and clinical medicine Vol. 165; no. 5; pp. 558 - 577
• Main Authors: Saha, Shilpi, Mukherjee, Shravanti, Mazumdar, Minakshi, Manna, Argha, Khan, Poulami, Adhikary, Arghya, Kajal, Kirti, Jana, Debarshi, Sa, Gaurisankar, Mukherjee, Sanhita, Sarkar, Diptendra K, Das, Tanya
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.05.2015
• Subjects: Adult; Aged; Antineoplastic Agents - therapeutic use; Antineoplastic Combined Chemotherapy Protocols; Apoptosis - drug effects; Breast Neoplasms - drug therapy; Breast Neoplasms - genetics; Breast Neoplasms - metabolism; Doxorubicin - therapeutic use; Drug Resistance, Neoplasm - drug effects; Female; Humans; Internal Medicine; MCF-7 Cells; Middle Aged; Neoplastic Stem Cells - drug effects; Neoplastic Stem Cells - metabolism; Neoplastic Stem Cells - pathology; Plicamycin - analogs & derivatives; Plicamycin - therapeutic use; Sp1 Transcription Factor - antagonists & inhibitors; Sp1 Transcription Factor - genetics; Spheroids, Cellular - drug effects; Spheroids, Cellular - pathology; Translational Research, Biomedical; Tumor Cells, Cultured; Tumor Microenvironment - drug effects; Tumor Microenvironment - genetics; NACT; XIAP; 5-fluorouracil, adriamycin, and cyclophosphamide; breast cancer stem cells; phosphate-buffered saline; ChIP; locally advanced breast cancer; X-linked inhibitor of apoptosis; chromatin immunoprecipitation; FITC; p53 upregulated modulator of apoptosis; LABC; reactive oxygen species; pH2AX; fluorescein isothiocyanate; PBS; peripheral blood mononuclear cells; dichlorofluorescein diacetate; bCSCs; hydrogen peroxide; FAC; phospho-H2A histone family, member X; chemotherapy resistant; PBMC; CR; chemotherapy sensitive; CS; H 2O 2; phycoerythrin; NAC; neoadjuvant chemotherapy; PE; ROS; DCF-DA; N-acetyl cysteine; PUMA; H2O2
• ISSN: 1931-5244; 1878-1810
• DOI: 10.1016/j.trsl.2014.10.011

Chemotherapy resistance is a major clinical challenge for the management of locally advanced breast cancer. Accumulating evidence suggests a major role of cancer stem cells (CSCs) in chemoresistance evoking the requirement of drugs that selectively target CSCs in combination with chemotherapy. Here, we report that mithramycin A, a known specificity protein (Sp)1 inhibitor, sensitizes breast CSCs (bCSCs) by perturbing the expression of drug efflux transporters, ATP-binding cassette sub-family G, member 2 (ABCG2) and ATP-binding cassette sub-family C, member 1 (ABCC1), survival factors, B-cell lymphoma 2 (Bcl-2) and X-linked inhibitor of apoptosis (XIAP), and, stemness regulators, octamer-binding transcription factor 4 (Oct4) and Nanog, which are inherently upregulated in these cells compared with the rest of the tumor population. In-depth analysis revealed that aberrant overexpression of Sp1 in bCSCs transcriptionally upregulates (1) resistance-promoting genes to protect these cells from genotoxic therapy, and (2) stemness regulators to sustain self-renewal potential of these cells. However, mithramycin A causes transcriptional suppression of these chemoresistant and self-renewal genes by inhibiting Sp1 recruitment to their promoters. Under such antisurvival microenvironment, chemotherapeutic agent doxorubicin induces apoptosis in bCSCs via DNA damage–induced reactive oxygen species generation. Cumulatively, our findings raise the possibility that mithramycin A might emerge as a promising drug in combinatorial therapy with the existing chemotherapeutic agents that fail to eliminate CSCs. This will consequently lead to the improvement of therapeutic outcome for the treatment-resistant breast carcinomas.