v‐src induces cisplatin resistance by increasing the repair of cisplatin–DNA interstrand cross‐links in human gallbladder adenocarcinoma cells

Activation of Src, which has an intrinsic protein tyrosine kinase (PTK) activity, has been demonstrated in human solid tumors, such as colorectal and breast cancers. To investigate the role of activated Src in drug resistance, we evaluated the effect of v‐src on the resistance to various anti‐cancer drugs using v‐src‐transfected HAG‐1 human gallbladder adenocarcinoma cells. Compared with parental or mock‐transfected HAG‐1 cells, v‐src‐transfected HAG/src3–1 cells showed a 3.5‐fold resistance to cis‐diamminedichloroplatinum (II) (CDDP) but not to doxorubicin, etoposide or 5‐fluorouracil. By contrast, activated H‐ras, which acts downstream of src, failed to induce resistance to either of these drugs. Furthermore, wortmannin, a phosphatidylinositol (PI) 3‐kinase inhibitor, and H7, a protein kinase C (PKC) inhibitor, did not alter CDDP resistance. Evaluation of the kinetics of the removal of DNA interstrand cross‐links (ICLs), measured by alkaline elution, showed a significant increase in this removal in HAG/src3–1 cells as compared with mock‐transfected cells, though no differences were found in the formation of DNA ICLs between these cell lines. CDDP resistance in v‐src‐transfected cells was reversed, if not completely, by either herbimycin A or radicicol, specific inhibitors of Src‐family PTKs, suggesting that Src tyrosine kinase activity induces CDDP resistance. Moreover, significant reduction in the repair of CDDP‐induced DNA ICLs was observed upon treatment with radicicol. The intracellular glutathione content and mRNA expression of topoisomerase II and metallothionein were virtually identical between these cell lines, except for topoisomerase I mRNA. Our data strongly suggest that the ability of activated src, but not ras, to induce CDDP resistance is mediated by augmentation of DNA repair through Src to downstream signal‐transduction pathways distinct from either the Ras, PI 3‐kinase or PKC pathway. Int. J. Cancer 80:731–737, 1999. © 1999 Wiley‐Liss, Inc.