April 2011
Volume 52, Issue 14
ARVO Annual Meeting Abstract  |   April 2011
ABCG2-Regulatory miRNAs And The Stem Cell Phenotype In Human Retinoblastoma Cells
Author Affiliations & Notes
  • Gail M. Seigel
    Center for Hearing and Deafness, SUNY at Buffalo; SUNY Eye Institute, Buffalo, New York
  • Yu-Zhuo Pan
    Department of Pharmaceutical Sciences,
    SUNY at Buffalo, Buffalo, New York
  • Zihua Hu
    Center for Excellence,
    SUNY at Buffalo, Buffalo, New York
  • Ai-Ming Yu
    Department of Pharmaceutical Sciences,
    SUNY at Buffalo, Buffalo, New York
  • Footnotes
    Commercial Relationships  Gail M. Seigel, None; Yu-Zhuo Pan, None; Zihua Hu, None; Ai-Ming Yu, None
  • Footnotes
    Support  NIH R21CA127061, U54CA143876, Research to Prevent Blindness (GMS); Interdisciplinary Research Development Fund (IRDF), SUNY Buffalo (AMY).
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 1595. doi:
  • Views
  • Share
  • Tools
    • Alerts
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Gail M. Seigel, Yu-Zhuo Pan, Zihua Hu, Ai-Ming Yu; ABCG2-Regulatory miRNAs And The Stem Cell Phenotype In Human Retinoblastoma Cells. Invest. Ophthalmol. Vis. Sci. 2011;52(14):1595.

      Download citation file:

      © ARVO (1962-2015); The Authors (2016-present)

  • Supplements

Purpose: : Retinoblastoma (RB) is a blinding retinal tumor of early childhood that is often treated with enucleation and/or chemotherapy/radiation therapies. In our previous studies, we have shown that there are small subpopulation(s) of RB cells with stem-like properties that include the expression of ABCG2, a stem cell marker and drug transporter that confers resistance to over 20 chemotherapy drugs. The goal of this study was to compare ABCG2+ and ABCG2- RB cells for the presence of three specific miRNAs that regulate the expression of ABCG2 as a possible target for molecular pharmacological interventions.

Methods: : We separated human RB143 cells into ABCG2+ and ABCG2- enriched populations by immunomagnetic positive selection and compared the two populations by RT-qPCR and miRNA-selective stem-loop RT qPCR. These methods were used to determine the expression of miRNAs, ABC transporters (MDR1/ABCB1, MRP1/ABCC1 and BCRP/ABCG2), as well as the expression of two additional stem cell markers ALDH1 and CD133.

Results: : : ABCG2 (normalized to 18S) was ~51-fold higher in ABCG2+ cells vs. ABCG2- cells, a strong validation of cell enrichment by the immunomagnetic positive selection method. As an additional confirmation of the stem cell phenotype, the stem cell markers ALDH1 and CD133 were ~8- and 10-fold higher in ABCG2+ cells, respectively. When we examined two other ABC transporters, there was no difference in ABCB1 expression between ABCG2- and ABCG2+ cells, while MRP1/ABCC1 was undetectable in both populations. We then analyzed miR-328, miR-519c and miR-520h, three miRNAs that are reported to regulate ABCG2 at the post-transcriptional level. Very interestingly, miR-328 (normalized to U74) was ~9-fold lower and miR-519c was ~15-fold lower in ABCG2+ cells. miR-520h was ~3-fold lower in ABCG2+ cells.

Conclusions: : In summary, ABCG2+ enriched cells demonstrate important differences from their ABCG2- counterparts in terms of ABCG2 miRNA regulatory molecules. Additional analysis of functionally relevant miRNAs may lead to new strategies in targeting ABCG2-mediated chemoresistance and metastasis in retinoblastoma, as well as in other malignancies.

Keywords: retinoblastoma • gene/expression • pathology: human 

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.