Reversal of cisplatin and multidrug resistance by ribozyme-mediated glutathione suppression

gamma -Glutamylcysteine synthetase (gamma -GCS) is a key enzyme in glutathione (GSH) synthesis, and is thought to play a significant role in intracellular detoxification, especially of anticancer drugs. Increased levels of GSH are commonly found in the drug-resistant human cancer cells. We designed a hammerhead ribozyme against gamma -GCS mRNA (anti-gamma -GCS Rz), which specifically downregulated gamma -GCS gene expression in the HCT-8 human colon cancer cell line. The aim of this study was to reverse the cisplatin and multidrug resistance for anticancer drugs. The cisplatin-resistant HCT-8 cells (HCT-8DDP cells) overexpressed MRP and MDR1 genes, and showed resistance to not only cisplatin (CDDP), but also doxorubicin (DOX) and etoposide (VP-16). We transfected a vector expressing anti-gamma -GCS Rz into the HCT-8DDP cells (HCT-8DDP/Rz). The anti-gamma -GCS Rz significantly suppressed MRP and MDR, and altered anticancer drug resistance. The HCT-8DDP/Rz cells were more sensitive to CDDP, DOX and VP-16 by 1.8-, 4.9-, and 1.5-fold, respectively, compared to HCT-8DDP cells. The anti-gamma -GCS Rz significantly down-regulated gamma -GCS gene expression as well as MRP/MDR1 expression, and reversed resistance to CDDP, DOX and VP-16. These results suggested that gamma -GCS plays an important role in both cisplatin and multidrug resistance in human cancer cells. (C) 2001 Academic Press.