Cellular and molecular determinants of all‐trans retinoic acid sensitivity in breast cancer: Luminal phenotype and RARα expression

• Published in: EMBO molecular medicine 2015-07, Vol.7 (7), p.950-972
• Main Authors: Centritto, Floriana, Paroni, Gabriela, Bolis, Marco, Garattini, Silvio Ken, Kurosaki, Mami, Barzago, Maria Monica, Zanetti, Adriana, Fisher, James Neil, Scott, Mark Francis, Pattini, Linda, Lupi, Monica, Ubezio, Paolo, Piccotti, Francesca, Zambelli, Alberto, Rizzo, Paola, Gianni', Maurizio, Fratelli, Maddalena, Terao, Mineko, Garattini, Enrico
• Format: Article
• Language: English
• Subjects: Animals; Antineoplastic Agents - pharmacology; Antineoplastic Agents - therapeutic use; Breast cancer; Breast Neoplasms - pathology; Carcinoma; Cell Line, Tumor; Cytotoxicity; Disease Models, Animal; EMBO02; EMBO03; ErbB-2 protein; Estrogen receptors; Experiments; Female; Gene Expression; Gene Expression Profiling; Gene Silencing; Genotype & phenotype; Humans; Ligands; luminal phenotype; Mammary gland; Menopause; Metastases; nuclear receptor; Phenotypes; RARalpha; Receptors, Retinoic Acid - biosynthesis; Research Article; Retinoic acid; Retinoic Acid Receptor alpha; Retinoic acid receptors; Transcription; Transplantation, Heterologous; Tretinoin - pharmacology; Tretinoin - therapeutic use; Tumor cell lines; Tumors; Xenografts
• ISSN: 1757-4676; 1757-4684
• DOI: 10.15252/emmm.201404670

Forty‐two cell lines recapitulating mammary carcinoma heterogeneity were profiled for all‐ trans retinoic acid (ATRA) sensitivity. Luminal and ER + (estrogen‐receptor‐positive) cell lines are generally sensitive to ATRA, while refractoriness/low sensitivity is associated with a Basal phenotype and HER2 positivity. Indeed, only 2 Basal cell lines ( MDA‐MB157 and HCC‐1599 ) are highly sensitive to the retinoid. Sensitivity of HCC‐1599 cells is confirmed in xenotransplanted mice. Short‐term tissue‐slice cultures of surgical samples validate the cell‐line results and support the concept that a high proportion of Luminal/ ER + carcinomas are ATRA sensitive, while triple‐negative ( Basal ) and HER2‐positive tumors tend to be retinoid resistant. Pathway‐oriented analysis of the constitutive gene‐expression profiles in the cell lines identifies RARα as the member of the retinoid pathway directly associated with a Luminal phenotype, estrogen positivity and ATRA sensitivity. RARα3 is the major transcript in ATRA‐sensitive cells and tumors. Studies in selected cell lines with agonists/antagonists confirm that RARα is the principal mediator of ATRA responsiveness. RARα over‐expression sensitizes retinoid‐resistant MDA‐MB453 cells to ATRA anti‐proliferative action. Conversely, silencing of RARα in retinoid‐sensitive SKBR3 cells abrogates ATRA responsiveness. All this is paralleled by similar effects on ATRA‐dependent inhibition of cell motility, indicating that RARα may mediate also ATRA anti‐metastatic effects. We define gene sets of predictive potential which are associated with ATRA sensitivity in breast cancer cell lines and validate them in short‐term tissue cultures of Luminal/ ER + and triple‐negative tumors. In these last models, we determine the perturbations in the transcriptomic profiles afforded by ATRA. The study provides fundamental information for the development of retinoid‐based therapeutic strategies aimed at the stratified treatment of breast cancer subtypes. Synopsis This study provides fundamental information for the development of retinoid‐based therapies for breast cancer and suggests that therapeutic strategies based on the use of RARα‐specific retinoids may overcome toxicity due to use of pan‐RAR agonists. Profiling of 42 breast cancer cell lines for ATRA sensitivity demonstrates that Luminal and ER + (estrogen receptor‐positive) cells are generally responsive, while Basal and HER2 + cells are generally refractory to the retinoid. The results o