Systematic Evaluation of Gastric Tumor Cell Index and Two-Drug Combination Therapy via 3-Dimensional High-Throughput Drug Screening

Abstract

Tumor heterogeneity greatly limits personalized treatment of cancer. Patient-derived tumor cell (PDC) models precisely recapitulate the molecular properties and biology of the disease, making them effective preclinical tools for assessing anti-cancer drug activities. Accurate estimation of tumor purity is essential for performing high-throughput drug screening (HTS). In the present study, we measured and predicted the tumor population index in PDC models for two-drug combinational strategies using HTS system. Gastric cancer cell-lines and PDCs were subjected to multi-color immunofluorescence analysis against EpCAM and vimentin to evaluate the tumor cell index based on EpCAM expression levels. We generated a tumor purity prediction model using five different gastric cancer cell-lines (AGS, KATO-III, MKN-45, NCI-N87, SNU-216) with fluorescence intensity-based techniques. Afterwards, stage IV gastric cancer PDC models were evaluated using a micropillar/microwell chip-based HTS system. HER2/CCNE1-amplified PDCs were considerably resistant to an HER2 inhibitor, while combinational treatment consisting of an HER2 inhibitor with anti-WEE1 compound substantially suppressed tumor cellular growth. Moreover, PDCs with BRCA1/2 mutations were synergistically sensitive to HER2 and PARP inhibition therapy. Finally, somatic mutations in TP53 and CDKN2A with MYC amplification rendered PDCs susceptible to the drug combination of WEE1 and HER2. Collectively, our systematic method of high-throughput drug sensitivity screening is an integral pre-clinical platform for evaluating potential two-drug combinational approaches for personalized treatment of cancer.

Purinergic P2Y2 and P2X4 Receptors Are Involved in the Epithelial-Mesenchymal Transition and Metastatic Potential of Gastric Cancer Derived Cell Lines

“…TEER experiments and molecular interventions revealed that, at non-transfected conditions, GC-derived cell lines had lower electrical resistance values than their nontumoral counterpart GES-1 (TEER ranging from 26 to 62 Ω cm 2 ), with the only exception of MKN-74 cells, which have noticeably higher TEER levels (from 140 to 230 Ω cm 2 ). This situation observed for AGS and MKN-45 cells are consistent with the high cell motility and EMT-induction profile defined by the aberrant nuclear vimentin distribution that has been described for both cell lines during a high-throughput analysis of gastric tumors for drug screening [56]. MKN-74 TEER levels can be explained considering their moderate differentiation grade, which gives them an epithelial-like phenotype, and their high P2Y2Rinduced proliferation rates [14], allowing them to reach 100% confluence after 24 h postseeding, giving them the electrical resistance pattern observed by us.…”

Construction and Evaluation of BAL-PTX Co-Loaded Lipid Nanosystem for Promoting the Anti-Lung Cancer Efficacy of Paclitaxel and Reducing the Toxicity of Chemotherapeutic Drugs

“…Combination index (CI) based on median effect analysis. 28 CI > 1, CI = 1, and CI < 1 indicate antagonism, addition, and synergy, respectively. 29 , 30 …”