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J. Biol. Chem., Vol. 280, Issue 25, 23987-24003, June 24, 2005

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Gene Expression Profiling to Identify Oncogenic Determinants of Autocrine Human Growth Hormone in Human Mammary Carcinoma^*

Xiu Qin Xu, B. Starling Emerald¶, Eyleen L. K. Goh||, Nagarajan Kannan¶, Lance D. Miller, Peter D. Gluckman¶, Edison T. Liu, and Peter E. Lobie¶**

From the Microarray and Expression Genomics, Genome Institute of Singapore, Republic of Singapore, ^¶Liggins Institute and the National Research Center for Growth and Development, University of Auckland, 2-6 Park Avenue, Private Bag 92019, Auckland, New Zealand and the ^||Institute of Molecular and Cell Biology, 30 Medical Drive, Singapore 117609, Republic of Singapore

We have exploited a discrepancy in the oncogenic potential of autocrine and exogenous human growth hormone (hGH) in an attempt to identify molecules that could potentially be involved in oncogenic transformation of the human mammary epithelial cell. Microarray analysis of 19,000 human genes identified a subset of 305 genes in a human mammary carcinoma cell line that were remarkably different in their response to autocrine and exogenous hGH. Autocrine and exogenous hGH also regulated 167 common genes. Semiquantitative reverse transcription-PCR confirmed differential regulation of genes by either autocrine or exogenous hGH. Functional analysis of one of the identified autocrine hGH-regulated genes, TFF3, determined that its expression is sufficient to support anchorage-independent growth of human mammary carcinoma cells. Small interfering RNA-mediated knockdown of TFF3 concordantly abrogated anchorage-independent growth of mammary carcinoma cells and abrogated the ability of autocrine hGH to stimulate oncogenic transformation of immortalized human mammary epithelial cells. Further functional characterization of the identified subset of specifically autocrine hGH regulated genes will delineate additional novel oncogenes for the human mammary epithelial cell.

Received for publication, April 11, 2005

^* This work was supported by the Marsden Fund Royal Society of New Zealand, the Foundation for Research, Science and Technology, and the National Research Center for Growth and Development. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Supported by the Agency for Science, Technology and Research (A^* Star) of Singapore.

^** To whom all correspondence should be addressed. Tel.: 64-9-3737599 (ext. 82125); Fax: 64-9-3737497; E-mail: p.lobie{at}auckland.ac.nz.

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