Circumvention of Drug Resistance in Cisplatin-resistant Sublines of the Human Squamous Carcinoma Cell Line HLac 79 in Vitro and in Vivo

Abstract

In a previous report we have characterized cisplatin (CDDP)-resistant sublines (HLac 79-DDP1 to DDP4) of the recloned squamous cell head and neck cancer (SCHNC) line HLac 79-ML revealing significant alterations of glutathione (GSH) metabolism and drug accumulation. in order to overcome CDDP-resistance in HLac 79 cells we now investigated the effect of buthionine sulfoximine (BSO), a specific inhibitor of GSH synthesis, verapamil (VRP), a calcium channel blocker that has been found to modulate resistance towards a broad spectrum of antineoplastic drugs, cyclosporin a (CSA), an immunosuppressive agent probably affecting drug pharmacokinetics, and aphidicolin (APC), a fungal metabolite interfering with DNA repair through inhibition of DNA polymerase alpha, on HLac 79 CDDP-sensitivity. Using the colorimetric MTT assay, GSH depletion with BSO led to a significant decrease of the 50 % inhibitory drug concentration (IC₅₀) in all HLac 79 sublines by dose modifying factors (IC₅₀, CDDP/IC₅₀ BSO + CDDP) ranging from 1.8 to 3.3. VRP, CSA or APC were not effective to overcome CDDP resistance in HLac 79 cells. the potential of BSO to modulate CDDP resistance in vitro was tested in vivo in HLac 79 tumor bearing NMRI nu-nu mice subsequently. Oral administration of BSO 7 days prior and during (days -7 to 8) CDDP treatment (3 mg/kg bw i.p. days 0, 4, 8) produced a significant prolongation of mean survival time x as compared to chemotherapy alone. This held true for both the maternal line ML in terms of chemosensitization (CDDP: x=40.2 ± 15.9 days vs. CDDP+BSO: x=80.3 ± 30.4 days, p<0.001) and the CDDP resistant subline DDP4 in terms of partially overcoming secondary drug resistance (CDDP: x=56.5 ± 13.6 days vs. CDDP+BSO: x=72.5 ± 15.8 days, p<0.001). Enhanced toxicity of combined BSO and CDDP treatment manifested by transient 10% reduction of animal mean body weight.

Key Words:

• buthionine sulfoximine
• nude mouse xenotransplants