Table 2 Summary of HSC for drug discovery from TCM
From: High content screening for drug discovery from traditional Chinese medicine
Models | Principle | Platform | Purpose | Identified compound | Refs. |
|---|---|---|---|---|---|
eIF4E immunostaining in MEF cells. | mTOR signaling pathway affects cellular localization of eIF4E | Array-Scan® VTI HCS Reader (Thermo Scientific, USA) | Discover novel mTOR signaling pathway inhibitors | 1,4-O-diferuloylsecoisola-riciresinol and PierreioneB | [31] |
Nuclear translocation of NFATc1 in EGFP-NFATc1 U2OS cells | NFATc1 nuclear translocation affects osteoclast genesis and bone erosion | A confocal microscopy (Zeiss 710) | Screen for drug candidate for relevant diseases caused by osteoclast genesis | 6-[10(Z)-heptadecenyl] salicylic acid from Syzygium tetragonum Wall | [32] |
GFP-LC3-expressing HeLa cells | Autophagy induce the number of GFP-LC3 puncta rapidly increases | Opera HCS System, Columbus2.3 Software (Perkin-Elmer) | Screen for novel apoptosis and autophagy regulators | Neobractatin isobractatin from Garcinia bracteata | [33] |
African green monkey kidney cell line BSC-1 | Microtubules are essential for proper chromosome congression and separation | Automated microscope Axiovert 200Â M (Carl Zeiss, Germany) data was analyzed using MetaMorph 7.7.8.0 (Visitron, Germany) | Search for inhibitor of mitosis | Podoverine A from the plant Podophyllum versipelle Hance | [34] |
Primary cortical neuron and dorsal root ganglion | Morphological change of synapse and the neurite outgrowth reflect neuron viability | ImageXpress Micro automated wide-field fluorescent microscope (Molecular Devices) | Discover potential neuroprotective drugs for paclitaxel-induced neurotoxicity | 5-Hydroxydecanoate (5-HD) | [35] |
Cell cycle or cytoskeletal stain on HeLa cells | Cell cycle and cytoskeleton is the markers for cell growth and stress | ImageXpress Micro inflorescent microscope (Molecular Devices) | Develop new tool to identify novel compound from large candidate libraries | Quinocinnolinomycins | [36] |
β-catenin staining in the human MG-63 OS cell line | β-catenin is the reporter of WNT signaling | (TFM-680; Shanghai Tuming Optical Instrument Co., Ltd.) | Identify WNT signaling inhibitor for the treatment of osteosarcoma | Resveratrol | [37] |
The human prostate cancer cell line DU145 | The position of lysosomes to the nucleus is a readout for lysosome anterograde trafficking | Cellomics Arrayscan (Thermo Fisher Scientific, Inc, Waltham, MA) | Screen for drugs that inhibit lysosome trafficking | Niclosamide | [38] |
Non-small cell lung cancer (NSCLC) cell lines NCI-H157 and NCI-H460 | Cell death can be monitored by Anexin V/PI and DAPI staining | Kinetic Scan Reader (ThermoFisher Scientific, USA) | Identify the compounds to induce cell death | Ophiopogonin B from Radix Ophiopogon Japonicus | [39] |
p-ERK and c-myc staining in MKP-1 overexpression cells | p-ERK and c-myc reflect MKP-1 activity | ArrayScan II (Cellomics, Pittsburgh, PA) S-Plus statistical Software package (Insightful, Inc., Seattle, WA) | Identify the inhibitor of MKP-1 | Sanguinarine | [40] |
Apoptotic markers staining | Apoptotic markers reflect cells death | KineticScan Reader (Cellomics Inc, Pittsburg, PA, USA) | To identify hepatoprotective compounds | Salvianolic acid B from Radix Salviae miltiorrhizae | [41] |
Neurite imaging of SH-SY5Y cells | Neurite outgrowth is a marker for ROCK2 inhibition | In Cell Analyzer 2000 (GE, Healthcare) IN Cell Investigator | Discover ROCK2 inhibitors | BIPM | [42] |
LX-2 hepatic stellate cell proliferation | Hepatic stellate cell proliferation is a marker of liver fibrosis | Cellomics Arra yScan VTI HCS Reader Cellomics Cell Health Profiling BioApplication | Identify the active components of Danshen for anti-liver fibrosis | Salvianolic acid B, caffeic acid and rosmarinic acid | [43] |
HepG2 cell | Apoptotic markers reflect cells death | Assay Scan VTI HCS Reader | Investigate the anti-proliferative and apoptotic effects of TMP | Tetramethylpyrazine | [44] |
A549 cells | Nuclei morphology reflects cell growth and mitosis status | BeckmanCoulter/Q3DM EIDAQ100, Activity Base and XLfit software | Identify compound that induces mitotic arrest | A novel quinazolinone | [45] |
Apoptotic markers staining on MCF7 cancer cells | Apoptotic markers reflect cells death | Eclipse TE2000-S microscope (Nikon, Tokyo, Japan) | Identify anti-cancer compounds | Koenimbin | [46] |
The NF-κB translocation staining in HT-29 cells | Nuclear translocation of NF-κB reflects inflammation | Cellomic ArrayScan HCS Reader (Thermo Scientific, USA) | Determine the anti-cancer and anti-inflammatory effects of PAB | Pseudolaric acid B (PAB) | [47] |
The NF-κB translocation staining in A549 lung cancer cells | Nuclear translocation of NF-κB reflects inflammation | ArrayScan HCS system (Cellomics), ArrayScan II Data Acquisition and Data Viewer version 3.0 (Cellomics) | Identify the NF-κB inhibitor | Panduratin A | [48] |
Rhodamine 123 staining of mitochondrial membrane potential in H9c2 cardiac muscle Cells | Mitochondrial membrane permeability change affect Rhodamine 123 signal | Leica DMI 6000 B (Leica Microsystems, Wetzlar, Germany) Cell Detection 1.0 Software | Identify compound to rescue mitochondrial membrane potential decrease | Salvianolic acid B, protocatechuic aldehyde, and tanshinone II A | [49] |
HeLa cells stably expressing EGFP-LC3 | GFP-LC3 is the marker for Autophagosome | In Cell 2000 system (GE Healthcare) | Find therapeutic method in chemotherapy of cancer by inhibiting autophagy | Dauricine and daurisoline | [50] |
Stable HeLa cells overexpressing 3×Flag-TFEB | TFEB nuclear translocation indicates biogenesis of autophagy-lysosome pathway | Opera high content system; Perkin-Elmer | Identify activators of TFEB | Curcumin derivative termed C1 | [51] |