TQ has also been shown to potentiate the anti-tumor AZD8931 in vivo activity GW3965 price of CDDP in Ehrlic ascites sarcoma (EAC) and simultaneously protected against CDDP nephrotoxicity [12]. Using both mouse and other rodent models it was shown that TQ when administered orally after mixing in drinking water ameliorated the nephrotoxicity from CDDP and also improved CDDP therapeutic index. Combining the most active chemotherapeutic drugs with agents that target specific pathways offers a powerful approach to cancer treatment and may counteract the many ways

that human cancer cells can become drug resistant. The platinum atom of CDDP forms covalent bonds to the N7 positions of purine bases to afford primarily 1, 2- or 1, 3-intra strand cross links and a lower number of inter strand cross links which eventually leads to apoptosis find more [13]. There is evidence that CDDP induces increased expression of NF-κB and that this activity results in increased CDDP resistance [14]. NF-κB controls cellular proliferation in part by increasing expression of cyclin

D1 which moves cells from G1 to S phase [15]. TQ has been reported to suppress tumor necrosis factor (TNF) induced NF-κB expression in human chronic myeloid leukemia cells (KBM-5) which may also explain why cells undergo apoptosis [16]. TQ was shown to suppress expression of NF-κB activation pathway through modulation of p65 subunit of NF-κB and inhibition of IκBα kinase (IKK) [16]. Thus in the present study we have combined a non-cell cycle specific Morin Hydrate active chemotherapy

drug CDDP which causes direct DNA damage with another agent TQ which targets the cell cycle at the transition from G1 to S phase hypothesizing the combination of TQ and CDPP will enhance the efficacy of CDDP and possibly overcome its resistance by suppression of CDDP induced over expression of NF-κB. TQ by suppressing NF-κB, should also affect tumor angiogenesis and metastasis [15] Materials and methods In Vitro experiments Cell culture NSCLC cell line NCI-H460 was generously provided by Dr James A. Cardelli (Louisiana State University Health Sciences Center, Shreveport, LA). SCLC cell line NCI-H146 was purchased from American Type Culture Collection (ATCC). Cells were grown in RPMI 1640 (Cell gro) supplemented with 10% Fetal bovine serum (FBS), 1% Penicillin and Streptomycin in a humidified incubator with 5% CO2 at 37°C. 1) Cell proliferation assay NCI-H460 cells (NSCLC cell line) were seeded at a density of 5,000 cells per well in 96 well plates and after 24 hrs cells were treated with 80 μM and 100 μM Thymoquinone (TQ) (Sigma Aldrich, St Louis MO) in 0.1% DMSO, 1.25 μM, 2.5 μM and 5.0 μM Cisplatin (CDDP) (Sigma Aldrich, St Louis MO) or TQ and CDDP at various combinations as noted. These doses of TQ and CDDP were chosen based on IC50 calculated from earlier experiments (Results not shown). There were four wells per condition and experiment was repeated twice to validate results.