Mir-302 reprograms human skin cancer cells into a pluripotent ES-cell-like state

Shi-Lung Lin; Donald C. Chang; Samantha Chang-Lin; Chun-Hung Lin; David T.S. Wu; David T. Chen; Shao-Yao Ying

doi:10.1261/rna.1162708

Mir-302 reprograms human skin cancer cells into a pluripotent ES-cell-like state

¹Department of Cell and Neurobiology, Keck School of Medicine, University of Southern California, Los Angeles, California 90033, USA
²Dental Department, Taiwan Adventist Hospital, Taipei City 10556, Taiwan, Republic of China
³Department of Ear, Nose and Throat, Tzu Chi General Hospital, Hualien 970, Taiwan, Republic of China

Abstract

Renewal of stem cells differs from cancer cell growth in self-controlled cell division. The mir-302 microRNA (miRNA) family (mir-302s) is expressed most abundantly in slow-growing human embryonic stem (ES) cells, and quickly decreases after cell differentiation and proliferation. Therefore, mir-302s was investigated as one of the key factors essential for maintenance of ES cell renewal and pluripotency in this study. The Pol-II-based intronic miRNA expression system was used to transgenically transfect the mir-302s into several human cancer cell lines. The mir-302 – transfected cells, namely, miRNA-induced pluripotent stem (mirPS) cells, not only expressed many key ES cell markers, such as Oct3/4, SSEA-3, SSEA-4 ,Sox2, and Nanog, but also had a highly demethylated genome similar to a reprogrammed zygotic genome. Microarray analyses further revealed that genome-wide gene expression patterns between the mirPS and human ES H1 and H9 cells shared over 86% similarity. Using molecular guidance in vitro, these mirPS cells could differentiate into distinct tissue cell types, such as neuron-, chondrocyte-, fibroblast-, and spermatogonia-like primordial cells. Based on these findings, we conclude that mir-302s not only function to reprogram cancer cells into an ES-like pluripotent state but also to maintain this state under a feeder-free cultural condition, which may offer a great opportunity for therapeutic intervention.

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Footnotes

Reprint requests to: Shi-Lung Lin, Department of Cell and Neurobiology, Keck School of Medicine, BMT-403, University of Southern California, 1333 San Pablo Street, Los Angeles, CA 90033; e-mail: lins{at}usc.edu; fax: (323) 442-3466; or Shao-Yao Ying, Department of Cell and Neurobiology, Keck School of Medicine, BMT-403, University of Southern California, 1333 San Pablo Street, Los Angeles, CA 90033; e-mail: sying{at}usc.edu; fax: (323) 442-3466.
Abbreviations: miRNA, microRNA; mir-302s, the mir-302 family; ES, embryonic stem; iPS, induced pluripotent stem; mirPS, mir-302-induced pluripotent stem; EB, embryoid body; CSC, cancerous stem cell; Pol-II, RNA polymerase type-II; NMD, nonsense-mediated decay.
Article published online ahead of print. Article and publication date are at http://www.rnajournal.org/cgi/doi/10.1261/rna.1162708.
- Received May 1, 2008.
- Accepted July 15, 2008.