ABT-737, a BH3 Mimetic, Enhances the Therapeutic Effects of Ionizing Radiation in K-ras Mutant Non-Small Cell Lung Cancer Preclinical Model.

PURPOSE: Tumor radioresistance and dose-limiting toxicity restrict the curative potential of radiotherapy, requiring novel approaches to overcome the limitations and augment the efficacy. Here, we investigated the effects of signal transducer and activator of transcription 3 (STAT3) activation and autophagy induction by irradiation on antiapoptotic proteins and the effectiveness of the BH3 mimetic ABT-737 as a radiosensitizer using K-ras mutant non-small cell lung cancer (NSCLC) cells and a KrasG12D:p53fl/fl mouse (KP mouse) model. MATERIALS AND METHODS: A549 and H460 cells were irradiated, and the expression of Bcl-2 family proteins, JAK/STAT transcriptional pathway, and autophagic pathway were evaluated by immunoblotting. The radiosensitizing effects of ABT-737 were evaluated using A549 and H460 cell lines with clonogenic assays and also by a KP mouse model with microcomputed tomography and immunohistochemistry. RESULTS: In A549 and H460 cells and mouse lung tissue, irradiation-induced overexpression of the antiapoptotic molecules Bcl-xL, Bcl-2, Bcl-w, and Mcl-1 through JAK/STAT transcriptional signaling induced dysfunction of the autophagic pathway. After treatment with ABT-737 and exposure to irradiation, the number of surviving clones in the cotreatment group was significantly lower than that in the group treated with radiation or ABT-737 alone. In the KP mouse lung cancer model, cotreatment with ABT-737 and radiation-induced significant tumor regression; however, body weight changes in the combination group were not significantly different, suggesting that combination treatment did not cause systemic toxicity. CONCLUSION: These findings supported the radiosensitizing activity of ABT-737 in preclinical models, and suggested that clinical trials using this strategy may be beneficial in K-ras mutant NSCLC.

Effects of saikosaponin A on cocaine self-administration in rats.

We have previously demonstrated that saikosaponin A (SSA) attenuated morphine self-administration behavior. In the present study, we evaluated the effects of SSA on cocaine-maintained responding using self-administration procedure. Rats self-administered cocaine (0.25mg/kg per infusion) under a fixed ratio 1 schedule of reinforcement during daily 3-h session. Once stable basal responses were obtained, rats were pretreated with each doses of SSA (1.0, 2.5, 5.0mg/kg) or its vehicle (5% Tween-80) by an intraperitoneal injection 30min before the start of self-administration testing. Additionally, different groups of rats received either the selective GABAB antagonist SCH 50911 or the GABAA antagonist bicuculline before systemic administration of SSA at dose of 2.5mg/kg. Results showed that SSA significantly reduced cocaine self-administration without affecting food consumption. SSA inhibition of cocaine reinforced-responding was blocked by SCH 50911, but not bicuculline. Results suggest that SSA may attenuate cocaine-reinforced behavior through activation of GABAB receptors.

Effect of saikosaponin A on maintenance of intravenous morphine self-administration.

In this study, we investigated the effects of saikosaponin A (SSA), a major compound of Bupleurum falcatum L., on morphine self-administration behavior. Male Sprague-Dawley rats were trained to self-administer intravenous morphine (0.1mg/kg per injection over 5s) during daily 1-h sessions under a fixed-ratio 1 schedule. Rats were pretreated with SSA (0.25, 0.5, 1.0mg/kg) by intraperitoneal injection 30 min prior to the start of the test session. Results demonstrated that pretreatment with SSA reduced morphine-maintained responding dose-dependently. Additionally, SSA inhibition of morphine-reinforced behavior was blocked by the selective GABA(B) receptor antagonist (2S)(+)-5,5-dimethyl-2-morpholineacetic acid (SCH 50911), but not the selective GABA(A) receptor antagonist bicuculline. Together, these results suggest that SSA may effectively suppress morphine-reinforced behavior by activating GABA(B) receptors.

Potential roles of GABA receptors in morphine self-administration in rats.

It is well established that the reinforcing effect of drugs of abuse is linked to the mesolimbic dopamine (DA) system. Morphine produces an increase in DA release in the brain, which may provide positive reinforcement contributing to the development of motivational aspects of drug-seeking and maintenance behavior. Several studies suggest that the GABA receptor system may play a significant role in the modulating the mesolimbic DA system. The purpose of this study was to investigate potential roles for GABA agonists in morphine self-administration behavior. Male Sprague-Dawley rats were trained to self-administrated morphine (0.1 mg/kg per injection) during daily 1-h sessions under a fixed ratio 1 schedule. Rats received an intravenous injection of the selective GABA(B) antagonist SCH 50911 (2.0 mg/kg) or an intraperitoneal injection of the GABA(A) antagonist bicuculline (1.0 mg/kg), immediately followed by either an intraperitoneal injection of baclofen (1.25 or 1.8 mg/kg) or muscimol (0.5 or 1.0 mg/kg, i.p.), 30 min prior to the start of test session. Results showed that pretreatment with baclofen or muscimol reduced morphine-maintenance response in a dose-dependent fashion and that baclofen and muscimol effects were reversed by injections of SCH 50911 and bicuculline, respectively. These data suggest that activation of both GABA(A) and GABA(B) receptors may be effective in suppressing the reinforcing effects of morphine.