Painting with a molecular brush: Genomic/proteomic interfacing to define the drug action profile of novel solid‐tumor selective anticancer agents

Abstract

XK469 is an investigational anticancer agent that exhibits antiproliferative activity in tumor-bearing animal models. We examined the drug-action profile of this agent at the molecular level regarding alterations induced in gene expression and proteins in HCT-116 human colon adenocarcinoma cells. We used a unique cDNA microarray (GeneMap TM Cancerarray) comprising 1152 human tumor-related genes and 2-D gel electrophoresis, respectively, following a 24-hour exposure to a drug concentration that killed a two-log fraction of HCT-116 clonogenic cells. Functional gene cluster profile (FGCP) analysis of the 71 out of 1152 genes that displayed a Ͼ2-fold increase or decrease in expression (over untreated control) identified a drug-specific involvement of the MAPK signal transduction pathway. MAPK signaling together with the involvement of ubiquitin proteins from 2-D gel electrophoresis suggest a novel drug-action profile at the molecular level for the in vitro antiproliferative activity of XK469. Cytometry 47: 72-79, 2002.

A Comparison of Drug-Treated and Untreated HCT-116 Human Colon Adenocarcinoma Cells Using a 2-D Liquid Separation Mapping Method Based upon Chromatofocusing PI Fractionation

“…A saturated matrix of CHCA in 50% acetonitrile with 0.1% TFA was freshly mixed. Angiotensin I, ACTH (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17), ACTH , and ACTH were added to the 1:4 diluted matrix solution and 0.5 µL of the resulting solution was spotted on a Micromass 96-spot plate. A 0.5-µL aliquot of the sample was then added on top of the spot, and the target was allowed to air-dry.…”

“…Many of these novel anticancer drugs in the developmental pipeline, however, exert their anti-proliferative activity through multiple targets that can be monitored using a "molecular brushing" approach combining cDNA microarrays and 2-D gel electrophoresis. 1 Since cellular proteins constitute functional gene expression and the latter proteomic technique has proven limited in its utility for pathway/target identification, better quantitative methods are clearly needed to more rapidly, accurately, and reproducibly perform comparative analyses of drug-induced alterations in both the protein expression and posttranslational modifications in drug-treated and untreated tumor cells. A robust technology that would meet the above prerequisites would prove invaluable to the ultimate discovery of novel protein targets responsible for the efficacy of unique anticancer agents with selectivity toward human antisolid tumor cells.…”

“…Thus, the discovery of novel drug compounds that directly inhibit selected protein targets is the focus of extensive research. Many of these novel anticancer drugs in the developmental pipeline, however, exert their antiproliferative activity through multiple targets that can be monitored using a “molecular brushing” approach combining cDNA microarrays and 2-D gel electrophoresis . Since cellular proteins constitute functional gene expression and the latter proteomic technique has proven limited in its utility for pathway/target identification, better quantitative methods are clearly needed to more rapidly, accurately, and reproducibly perform comparative analyses of drug-induced alterations in both the protein expression and posttranslational modifications in drug-treated and untreated tumor cells.…”

Kinetic study on the reaction of 2,4‐ and 2,6‐tolylene diisocyanate with 1‐butanol in the presence of styrene, as a model reaction for the process that yields interpenetrating polyurethane–polyester networks

“…This structure‐specific biological activity seemed particularly useful in identifying the proteins that are affected differently by the compounds and may control the initial phase of apoptotic cell death. In search of the prime molecular targets for apoptosis initiation in preneoplastic NCOL‐1 cells, we used a proteomic approach that enables us to delineate the action profile of novel tumor selective anticancer agents13 and identify new targets for intervention or markers of response 14. At this time, proteome analysis has only been applied to human colonic cells for identifying proteins that differ in expression level between normal and transformed tissues15 or that are altered by the treatment of cells with butyrate 16…”

Identification of biomarkers for the initiation of apoptosis in human preneoplastic colonocytes by proteome analysis

“…This structure-specific biological activity seemed particularly useful in identifying the proteins that are affected differently by the compounds and may control the initial phase of apoptotic cell death. In search of the prime molecular targets for apoptosis initiation in preneoplastic NCOL-1 cells, we used a proteomic approach that enables us to delineate the action profile of novel tumor selective anticancer agents 13 and identify new targets for intervention or markers of response. 14 At this time, proteome analysis has only been applied to human colonic cells for identifying proteins that differ in expression level between normal and transformed tissues 15 or that are altered by the treatment of cells with butyrate.…”