PMID- 11830524 OWN - NLM STAT- MEDLINE DCOM- 20020305 LR - 20171116 IS - 0008-5472 (Print) IS - 0008-5472 (Linking) VI - 62 IP - 3 DP - 2002 Feb 1 TI - Identification of molecular targets associated with selenium-induced growth inhibition in human breast cells using cDNA microarrays. PG - 708-14 AB - Past research indicated that methylseleninic acid (MSA) is an excellent tool for investigating the cancer chemopreventive action of selenium in vitro. The present study was designed to examine the cellular and molecular effects of MSA in the MCF10AT1 and MCF10AT3B premalignant human breast cells. After exposure to MSA, both cell lines exhibited a dose- and time-dependent growth-inhibitory response as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell proliferation assay. Further characterization of cellular and molecular changes was carried out only with the MCF10AT1 cells. Flow cytometry analysis showed that MSA blocked cell cycle progression at the G(0)-G(1) phase. Induction of apoptosis was also observed with the use of either the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) or the annexin V binding method. cDNA microarray analyses with cell cycle- and apoptosis-targeted arrays were then applied to profile the gene expression changes mediating these two cellular events. The analyses were conducted at 6 and 12 h of MSA treatment using synchronized cells. The expression signals of 30 genes were found to be significantly altered by MSA. These genes fall into three categories: cell cycle checkpoint controllers (e.g., cyclins, cdcs, cdks, E2F family proteins, and serine/threonine kinases), apoptosis regulatory genes (e.g., Apo-3, c-jun, and cdk5/cyclin D1), and signaling molecules [e.g., mitogen-activated protein (MAP)/extracellular signal-regulated protein kinase (ERK) and phosphatidylinositol 3'-kinase (PI3k) cascade genes]. The expression changes of 15 genes were selected for verification by Western or semiquantitative reverse transcription-PCR analyses. An agreement rate of 60% (9 of 15) was obtained from these confirmation experiments. On the basis of the above findings, tentative signaling pathways mediating the outcome of selenium-induced cell cycle arrest and apoptosis are proposed. The present study thus demonstrated the feasibility of applying cDNA microarray technology in delineating the mechanisms of the action of selenium and in pinpointing molecular targets as potential biomarkers for evaluating the efficacy of selenium intervention. FAU - Dong, Yan AU - Dong Y AD - Department of Experimental Pathology, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY 14263, USA. FAU - Ganther, Howard E AU - Ganther HE FAU - Stewart, Carleton AU - Stewart C FAU - Ip, Clement AU - Ip C LA - eng GR - CA 16056/CA/NCI NIH HHS/United States GR - CA 27706/CA/NCI NIH HHS/United States GR - CA 45164/CA/NCI NIH HHS/United States PT - Journal Article PT - Research Support, U.S. Gov't, P.H.S. PL - United States TA - Cancer Res JT - Cancer research JID - 2984705R RN - 0 (Anticarcinogenic Agents) RN - 0 (Growth Inhibitors) RN - 0 (Organoselenium Compounds) RN - 9900C6V162 (methylselenic acid) SB - IM MH - Anticarcinogenic Agents/*pharmacology MH - Apoptosis/drug effects MH - Breast/cytology/*drug effects/physiology MH - Breast Neoplasms/genetics/pathology/*prevention & control MH - Cell Cycle/drug effects MH - Cells, Cultured MH - G1 Phase/drug effects MH - Gene Expression/drug effects MH - Growth Inhibitors/pharmacology MH - Humans MH - Oligonucleotide Array Sequence Analysis MH - Organoselenium Compounds/*pharmacology MH - Resting Phase, Cell Cycle/drug effects EDAT- 2002/02/07 10:00 MHDA- 2002/03/07 10:01 CRDT- 2002/02/07 10:00 PHST- 2002/02/07 10:00 [pubmed] PHST- 2002/03/07 10:01 [medline] PHST- 2002/02/07 10:00 [entrez] PST - ppublish SO - Cancer Res. 2002 Feb 1;62(3):708-14.