PMID- 18593893 OWN - NLM STAT- MEDLINE DCOM- 20080805 LR - 20211020 IS - 1538-7445 (Electronic) IS - 0008-5472 (Print) IS - 0008-5472 (Linking) VI - 68 IP - 13 DP - 2008 Jul 1 TI - Single-cell transcription site activation predicts chemotherapy response in human colorectal tumors. PG - 4977-82 LID - 10.1158/0008-5472.CAN-07-6770 [doi] AB - Candidate gene and pathway approaches, and unbiased gene expression profiling, have identified marker signatures predictive of tumor phenotypes, such as drug sensitivity and invasive or metastatic potential. However, application of such information to evaluation of tumors in the clinic is limited by cell heterogeneity in the tumor. We have developed a novel method of fluorescence in situ hybridization (FISH) that can detect transcriptional activation of individual genes at their site in single cells in the interphase nucleus. A major obstacle in the treatment of colorectal cancer is relative insensitivity to the chemotherapeutic agent 5-Fluorouracil (5-FU). Here, we have developed a sensitive approach to predict relative sensitivity of colorectal cancer cells to 5-FU, using FISH with probes targeted to nascent mRNAs to measure the number of individual cells with active transcription sites for a panel of candidate genes. These results reveal that the transcriptional status of four key genes, thymidylate synthase (TYMS), MORF-related gene X (MRGX), Bcl2-antagonist/killer (BAK), and ATPase, Cu(2+) transporting beta polypeptide (ATP7B), can accurately predict response to 5-FU. As proof of principle, we show that this transcriptional profile is predictive of response to 5-FU in a small number of patient colon tumor tissues. This approach provides a novel ability to identify and characterize unique minor cell populations in the tumor that may exhibit relative resistance to chemotherapy. FAU - Pezo, Rossanna C AU - Pezo RC AD - Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Albert Einstein Cancer Center, Bronx, New York 10461, USA. FAU - Gandhi, Saumil J AU - Gandhi SJ FAU - Shirley, L Andrew AU - Shirley LA FAU - Pestell, Richard G AU - Pestell RG FAU - Augenlicht, Leonard H AU - Augenlicht LH FAU - Singer, Robert H AU - Singer RH LA - eng GR - R33CA83208/CA/NCI NIH HHS/United States GR - R33 CA083208/CA/NCI NIH HHS/United States GR - R33 CA083208-07/CA/NCI NIH HHS/United States GR - U54 CA100926-05/CA/NCI NIH HHS/United States GR - R01CA107382/CA/NCI NIH HHS/United States GR - R01CA70896/CA/NCI NIH HHS/United States GR - P30 CA013330/CA/NCI NIH HHS/United States GR - R01 CA107382/CA/NCI NIH HHS/United States GR - R01 CA075503/CA/NCI NIH HHS/United States GR - R33 CA083208-05A1/CA/NCI NIH HHS/United States GR - R33 CA083208-06/CA/NCI NIH HHS/United States GR - U54 CA100926-04/CA/NCI NIH HHS/United States GR - P30CA56036/CA/NCI NIH HHS/United States GR - U54 CA100926-03/CA/NCI NIH HHS/United States GR - R01CA75503/CA/NCI NIH HHS/United States GR - T32GM07288/GM/NIGMS NIH HHS/United States GR - R01 CA070896/CA/NCI NIH HHS/United States GR - P30CA13330/CA/NCI NIH HHS/United States GR - U54CA100926/CA/NCI NIH HHS/United States GR - R01CA86072/CA/NCI NIH HHS/United States GR - T32 GM007288/GM/NIGMS NIH HHS/United States GR - R01 CA086072/CA/NCI NIH HHS/United States GR - P30 CA056036/CA/NCI NIH HHS/United States GR - U54 CA100926/CA/NCI NIH HHS/United States PT - Evaluation Study PT - Journal Article PT - Research Support, N.I.H., Extramural PL - United States TA - Cancer Res JT - Cancer research JID - 2984705R RN - 0 (Antineoplastic Agents) RN - 0 (BAK1 protein, human) RN - 0 (Biomarkers, Pharmacological) RN - 0 (Cation Transport Proteins) RN - 0 (MORF4L2 protein, human) RN - 0 (Transcription Factors) RN - 0 (bcl-2 Homologous Antagonist-Killer Protein) RN - EC 2.1.1.45 (Thymidylate Synthase) RN - EC 3.6.1.- (Adenosine Triphosphatases) RN - EC 7.2.2.8 (ATP7B protein, human) RN - EC 7.2.2.8 (Copper-Transporting ATPases) RN - U3P01618RT (Fluorouracil) SB - IM MH - Adenosine Triphosphatases/analysis/genetics MH - Algorithms MH - Antineoplastic Agents/therapeutic use MH - Biomarkers, Pharmacological/analysis MH - Carcinoma/*diagnosis/*drug therapy/genetics MH - Cation Transport Proteins/analysis/genetics MH - Cell Line, Tumor MH - Colorectal Neoplasms/*diagnosis/*drug therapy/genetics MH - Copper-Transporting ATPases MH - Drug Resistance, Neoplasm/*genetics MH - Fluorouracil/*therapeutic use MH - HCT116 Cells MH - Humans MH - In Situ Hybridization, Fluorescence MH - Predictive Value of Tests MH - Prognosis MH - Thymidylate Synthase/analysis/genetics MH - Transcription Factors/analysis/genetics MH - Transcription Initiation Site/*physiology MH - bcl-2 Homologous Antagonist-Killer Protein/analysis/genetics PMC - PMC3107669 MID - NIHMS296021 COIS- Disclosure of Potential Conflicts of Interest L.H. Augenlicht: consultant/advisory board, Pittsburgh Cancer Center, Arizona Cancer Center, and Valley Hospital, New Jersey. R.H. Singer: consultant/advisory board, Aureon Laboratories. The other authors disclosed no potential conflicts of interest. EDAT- 2008/07/03 09:00 MHDA- 2008/08/06 09:00 PMCR- 2011/06/03 CRDT- 2008/07/03 09:00 PHST- 2008/07/03 09:00 [pubmed] PHST- 2008/08/06 09:00 [medline] PHST- 2008/07/03 09:00 [entrez] PHST- 2011/06/03 00:00 [pmc-release] AID - 68/13/4977 [pii] AID - 10.1158/0008-5472.CAN-07-6770 [doi] PST - ppublish SO - Cancer Res. 2008 Jul 1;68(13):4977-82. doi: 10.1158/0008-5472.CAN-07-6770.