Synergistic action by multi-targeting compounds produces a potent compound combination for human NSCLC both in vitro and in vivo

Abstract

By screening a collection of one hundred combinations of thiazolidinone compounds, we identified one combination (M4) that synergistically inhibited the growth of H460 and H460/TaxR cells and tumor growth in H460/TaxR xenograft mice. A whole genome microarray assay showed that genes involved in negative regulation of microtubule polymerization or depolymerization, intracellular protein kinase cascade, positive regulation of histone acetylation, cell cycle arrest and apoptosis were upregulated. Further analysis proved that the four compounds act as either microtubule polymerization inhibitors or histone deacetylase inhibitors. They act synergistically targeting multiple proteins and leading to the regulation of cell cycle checkpoint proteins, including p53, p21, cdc25C and cdc2, the activation of caspases, JNK, p38 cascades and the inactivation of Akt. These events resulted in the G2/M cell cycle arrest and cell apoptosis. These data provide a new strategy for discovering anticancer drugs and drug combinations for drug-resistant cancers.

From fighting depression to conquering tumors: a novel tricyclic thiazepine compound as a tubulin polymerization inhibitor

“…To explore the possible targets of TBPT, we determined the global gene expression profile of H460 TaxR cells after TBPT treatment using an Agilent whole-human genome oligo microarray platform. 20 The microarray analysis revealed that 534 genes were suppressed and 410 genes were upregulated (≥2-fold) in H460 TaxR cells following TBPT treatment (2 μ M, 4 h). The gene ontology analysis indicated that genes involved in regulating microtubule cytoskeleton and nucleus functions were altered by TBPT ( Supplementary Table S2 ).…”

“…15 , 16 , 17 In an effort to redeploy anti-depressant compound scaffold for anticancer applications, we synthesized a series of 27 novel tricyclic thiazepine derivatives 18 and screened these compounds against the NSCLC cell line H460 and its drug-resistant variant H460 TaxR (manuscript submitted). 19 , 20 , 21 The drug-resistant H460 TaxR cell line was obtained by treating H460 cells initially with 5 nM PTX and increased doses up to 100 nM, leading to a remarkable overexpression of P-gp (245-folds, Figure 1b ) among other possible alterations. Compared with H460, H460 TaxR is highly resistant to many clinical drugs, including PTX, vincristine and doxorubicin, because of the drug pump P-gp ( Figure 1c , Supplementary Table S1 ).…”

“…NSCLC xenograft tumor models were constructed as reported previously. 20 , 21 Four-week-old female athymic nude mice were purchased from Vital River Laboratory Animal Technology Co., Ltd (Beijing, China) and housed under pathogen-free conditions. This study was performed following the recommendations from the Guide for the Care and Use of Laboratory Animals by the National Institutes of Health.…”

“…RNA preparation and microarray hybridization were performed according to the manufacturer's instructions (Kangchen Bio-Tech, Shanghai, China) for the Agilent Whole Human Genome Oligo Microarray Platform (Santa Clara, CA, USA). 20 …”

Comprehensive Two- and Three-Dimensional RNAi Screening Identifies PI3K Inhibition as a Complement to MEK Inhibitor AS703026 for Combination Treatment of Triple-Negative Breast Cancer

“…Cells were treated with the drug combination and analyzed using a CellTiter-Blue assay (Promega) according to the manufacturer's instructions. Drug synergy was determined by the isobologram and combination-index methods using CalcuSyn software 2.1 (Biosoft) 59 . Detailed methods are described in the Supplementary Information.…”