TETRACYCLINE-INDUCIBLE PANCREATIC CANCER CELL LINES

Abstract

Precise control of the level of protein expression in cells can yield quantitative and temporal information on the role of a given gene. The most promising and widely used technique for regulating heterologous gene expression has been the tetracycline-regulated gene expression system. The technique consists of consecutive transfections of the target cells with two transgenes, the first of which encodes a reverse tetracycline-controlled transactivator protein (rtTA) and the second of which is the modified gene of interest, with a tetracycline response element (TRE) in its 5′ regulatory sequence. Although a second-generation tet-on transactivator was recently described, it has not been widely investigated for its potential as a tool for regulating genes in cells and particularly in cells previously recalcitrant to the first-generation tet-on approach, such as pancreatic cancer cells. Using this technique, four pancreatic cancer cell clones (derived from the pancreatic cancer cell line Suit-2) incorporating a second-generation doxycycline-inducible gene expression system have been developed. These four clones were tested for transient inducible expression of the luciferase gene and are currently being tested for stable inducible expression of the actin capping protein CapG. In conclusion, we have generated pancreatic cancer-derived cell clones in which expression of genes is potentially controllable.