Piperlongumine inhibits proliferation and oncogenic MYCN expression in chemoresistant metastatic retinoblastoma cells directly and through extracellular vesicles

Cui Shi; Kunhui Huang; John Soto; Renuka Sankaran; Vrinda Kalia; Onyekwere Onwumere; Michael Young; Linda Einbond; Stephen Redenti

doi:10.1016/j.biopha.2023.114554

Piperlongumine inhibits proliferation and oncogenic MYCN expression in chemoresistant metastatic retinoblastoma cells directly and through extracellular vesicles

Biomed Pharmacother. 2023 May:161:114554. doi: 10.1016/j.biopha.2023.114554. Epub 2023 Mar 20.

Authors

Cui Shi¹, Kunhui Huang¹, John Soto², Renuka Sankaran¹, Vrinda Kalia³, Onyekwere Onwumere⁴, Michael Young⁵, Linda Einbond², Stephen Redenti⁶

Affiliations

¹ Lehman College, the City University of New York, 250 Bedford Park Boulevard West, Bronx, NY 10468, USA; Biochemistry Doctoral Program, The Graduate School, City University of New York, 365 Fifth Avenue, New York, NY 10016, USA.
² Lehman College, the City University of New York, 250 Bedford Park Boulevard West, Bronx, NY 10468, USA.
³ Department of Environmental Health Sciences, Columbia University Mailman School of Public Health, New York, NY 10032, USA.
⁴ Lehman College, the City University of New York, 250 Bedford Park Boulevard West, Bronx, NY 10468, USA; Biology Doctoral Program, The Graduate School of the City University of New York, 365 5th Avenue, New York, NY 10016, USA.
⁵ The Schepens Eye Research Institute, Massachusetts Eye and Ear, Harvard Medical School, 20 Staniford Street, Boston, MA 02114, USA.
⁶ Lehman College, the City University of New York, 250 Bedford Park Boulevard West, Bronx, NY 10468, USA; Biochemistry Doctoral Program, The Graduate School, City University of New York, 365 Fifth Avenue, New York, NY 10016, USA; Biology Doctoral Program, The Graduate School of the City University of New York, 365 5th Avenue, New York, NY 10016, USA. Electronic address: Stephen.Redenti@lehman.cuny.edu.

Abstract

Ocular retinoblastoma malignancies, which develop into metastatic phenotypes, result in poor prognosis and survival for infant and child patients. To improve the prognosis of metastatic retinoblastoma, it is important to identify novel compounds with less toxic side effects and higher therapeutic efficacy compared to existing chemotherapeutics. Piperlongumine (PL), a neuroprotective, plant-derived compound has been explored for its anticancer activities both in vitro and in vivo. Here, we analyze the potential efficacy of PL for metastatic retinoblastoma cell treatment. Our data reveal that PL treatment significantly inhibits cell proliferation in metastatic retinoblastoma Y79 cells compared to the commonly used retinoblastoma chemotherapeutic drugs carboplatin, etoposide, and vincristine. PL treatment also significantly increases cell death compared to treatment with other chemotherapeutic drugs. PL-induced cell-death signaling was associated with significantly higher caspase 3/7 activities and greater loss of mitochondrial membrane potential. PL was also internalized into Y79 cells with an estimated concentration of 0.310pM and expression analysis revealed reduced MYCN oncogene levels. We next examined extracellular vesicles derived from PL-treated Y79 cells. Extracellular vesicles in other cancers are pro-oncogenic, mediating systemic toxicities via the encapsulation of chemotherapeutic drugs. Within metastatic Y79 EV samples, an estimated PL concentration of 0.026pM was detected. PL treatment significantly downregulated Y79 EV cargo of the oncogene MYCN transcript. Interestingly, non-PL-treated Y79 cells incubated with EVs from PL-treated cells exhibited significantly reduced cell growth. These findings indicate that in metastatic Y79 cells, PL exhibits potent anti-proliferation effects and oncogene downregulation. Importantly, PL is also incorporated into extracellular vesicles released from treated metastatic cells with measurable anti-cancer effects on target cells at a distance from the site of primary treatment. The use of PL in the treatment of metastatic retinoblastoma may reduce primary tumor proliferation and inhibit metastatic cancer activity systemically via extracellular vesicle circulation.

Keywords: Chemoresistance; Chemotherapeutic drugs; Extracellular vesicles; MYCN; Metastatic retinoblastoma; Piperlongumine.

MeSH terms

Cell Line, Tumor
Cell Proliferation
Humans
N-Myc Proto-Oncogene Protein / genetics
Retinal Neoplasms* / genetics
Retinoblastoma* / genetics

Substances

N-Myc Proto-Oncogene Protein
piperlongumine
MYCN protein, human

Grants and funding

SC3 GM113782/GM/NIGMS NIH HHS/United States