Lymphokine-activated killer cell susceptibility in epirubicin-resistant and parental human non-small cell lung cancer (NSCLC)

Abstract

The aim of this study was to evaluate that: (i) epirubicin-HCl (EPI) and lymphokine-activated killer (LAK) cells cytotoxicity may be mediated by free radical generation; and (ii) resistant H1299 cells may be more sensitive to combined treatment of LAK cells plus EPI than the LAK or EPI treatment alone. Viability of H1299 cells treated with EPI, LAK and LAK plus EPI was measured using the MTT test. Amount of glutathione (GSH), protein content and enzymatic activity were measured by spectrophotometer. Glutathione S-transferase (GST)-pi expression in the cells was determined by western blot analysis. LAK plus EPI combined treatment increased susceptibility of H1299 WT and H1299 EPI(R) (300-fold EPI resistant) cells to LAK cell cytotoxicity. The resistance of H1299 EPI(R) cells to EPI appears to be associated with a developed tolerance to free radicals, most likely because of a 2-fold increase in NADPH-dependentcytochrome-P450 reductase (NADPH-CYP reductase) activity, 11-fold GST activity and 11-and 7-fold augmented selenium dependent and independent glutathione peroxidase (GSH-Px) activity, respectively. Amount of GST-pi in H1299 EPI(R) cells is statistically different from negative control and H1299 WT (p < 0.01). It is proposed that production of reactive oxygen species and hydrogen peroxide by the treatment of EPI and LAK cells can cause cytotoxicity of H1299 WT and H1299 EPI(R) cells. Superoxide dismutase, catalase, GSH-Px, GST, NADPH-CYP reductase and GSH must be considered as part of the intracellular antioxidant defense mechanism of H1299 WT and H1299 EPI(R) cells against reactive oxygen species. Combined treatment of EPI plus LAK cells caused the increasing cytotoxicity on the H1299 EPI(R) cells.

Membrane and DNA Damaging/Protective Effects of Eugenol, Eucalyptol, Terpinen-4-ol and Camphor at Various Concentrations on Parental and Drug Resistant H1299 Cells

“…Cells were used when monolayer confluence had reached 75%. The drug-resistant (epirubicin-resistant) H1299 tumor cells were derived from the parental line by stepwise selection in increasing concentrations of epirubicin until the cells were capable of propagating in 220 ng/mL drug, as described previously (15,18).…”

“…A group of drug-resistant cells can develop in tumors during the chemotherapy. In one study, the activities of NADPH-CYP reductase (2-fold), GST (11-fold), Se-dependent and independent-GSH-Px (7-to 11-fold), and GST-pi were found higher in epirubicin-resistant H1299 cells than parental cells (15). Lung tumor cells selected for acquired resistance to epirubicin in cultures have concurrently developed a tolerance to superoxide and hydrogen peroxide, most likely because of elevated activities of enzymatic defenses against oxyradicals.…”

Evaluation of cytotoxic, membrane, and DNA damaging effects of Thymus revolutus Célak essential oil on different cancer cells

“…One study showed that drug-resistant cells could resist oxidants with their higher antioxidant enzymes (33). Our previous study showed that epirubicin-resistant H1299 cells had more GPx and GST activity, but lower GSH amounts, than parental H1299 cells (18). One study suggested that localization of EGFR is related to the sensitivity/resistance of cells to chemotherapy and radiotherapy.…”

“…Cells were used when monolayer confluence reached 75%. Epirubicin-resistant (drug-resistant) H1299 tumor cells were derived from the parental line by stepwise selection in increasing concentrations of epirubicin until the cells were capable of propagating in 220 ng/mL drug, as described previously (17,18).…”

A comparative study of cytotoxic, membrane and DNA damaging effects of Origanum majorana’s essential oil and its oxygenated monoterpene component linalool on parental and epirubicin-resistant H1299 cells

“…Acquired multidrug resistance is the main obstacle for the cure of SCLC. Increased antioxidant mechanisms enzymes amounts in drug resistance cells can be one of the reasons of acquired multidrug resistance (Ozkan 2007). A group of drug resistance cells can occur in tumours during the chemotherapy.…”