Screening Method for Anti-Colon Cancer Drugs Using Two Sensor Cell Lines with Human beta 4-Galactosyltransferase 4 Gene Promoters

摘要

The increased expression of beta 4-galactosyltransferase (beta 4GalT) 4 is closely associated with poor prognosis of colon cancer. Recently, we showed that the expression of the beta 4GalT4 gene is regulated by the 0.17 kb core promoter region containing one binding site for Specificity protein 1 (Sp1). To develop a screening method for anti-colon cancer drugs, two sensor cell lines having the luciferase gene under the control of two beta 4GalT4 gene promoters that differed in length were established from SW480 human colon cancer cells. The hGT4-0.17-sensor cells possessed the luciferase reporter driven by the 0.17 kb promoter, while the hGT4-0.3-sensor cells possessed the luciferase reporter driven by the 0.3 kb promoter containing one binding site each for colon cancer-related transcription factors including activator protein 2, E2F, caudal-related homeobox transcription factors, and Runt-related transcription factors besides Sp1. Upon treatment with mitogen-activated protein kinase signaling inhibitor U0126, the promoter activities of the hGT4-0.3-sensor cells decreased significantly, while those of the hGT4-0.17-sensor cells remained unchanged. These results suggest that the responsiveness to U0126 differs between two sensor cell lines due to the different regulation of the luciferase reporters. This study provides the screening method for anti-colon cancer drugs by the combination of two sensor cell lines.