Estimating Dynamic Cellular Morphological Properties Via The Combination Of The RTCA System And A Hough-Transform-Based Algorithm

Background: The x CELLigence real-time cell analysis(RTCA)system has the potential to detect cellular proliferation,migration,cytotoxicity,adherence,and remodeling.Although the RTCA system is widely recognized as a noninvasive and efficient tool for real-time monitoring of cellular fate,it cannot describe detailed cell morphological parameters,such as length and intensity.Methods: Transforming growth factor beta(TGF-β)induced the epithelial?mesenchymal transition(EMT),which produces significant changes in cellular morphology,so we used TGF-? to treat A549 epithelial cells in this study.We compared it with lipopolysaccharide(LPS)and cigarette smoke extract(CSE)as stimulators.We developed an efficient algorithm to quantify the morphological cell changes.This algorithm is comprised of three major parts: image preprocessing,Hough transform(HT),and post-processing.We used the RTCA system to record the A549 cell index.Western blot was used to confirm the EMT.Results: The RTCA system showed that different stimulators produce different cell index curves.The algorithm determined the lengths of the detected lines of cells,and the results were similar to the RTCA system in the TGF-? group.The Western blot results show that TGF-? changed the EMT markers,but the other stimulators remained unchanged.Conclusion: Optics-based computer vision techniques can supply the requisite information for the RTCA system based on good correspondence between the results.Background: Oral squamous cell carcinoma(OSCC)is one of the common malignant tumors in oral and maxillofacial region.With the high fatality,finding the treatment of OSCC becomes so urgent.Molecular targeted therapy has attracted much attention in recent years.At the same time,the network pharmacology provides an efficient technology approach to screen the putative targets of drugs.Huanglianjiedu Decoction(HLJDD)is a well-known traditional Chinese medicinal prescription,which consists of Coptidis rhizoma,Scutellariae radix,Phellodendri cortex and Gardeniae fructus.HLJDD has been used to the treatment of wide range of disease conditions.However,there is few research about HLJDD worked on OSCC.Therefore,we try to explore the mechanism of HLJDD on OSCC based on the network pharmacology.Methods: Using Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),we searched and screened the potential components of HLJDD and their putative targets.Then we found the potential targets of OSCC from Therapeutic Target Database(TTD),Online Mendelian Inheritance in Man(OMIM)and so on.We draw the protein-protein interaction(PPI)network by Cytoscape v3.2.0.With the topology analysis,we got the core targets from HLJDD potential active ingredients related targets-OSCC related targets PPI network.Moreover,we used the DAVID website and Cytoscape v3.2.0 software to do the Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis.Based on the results of enrichment,we checked a part of biological process(cell cycle,apoptosis and proliferation)and pathways on SCC-25 and CAL-27 cell lines.Results: The related potential components targets number of Coptidis rhizoma,Scutellariae radix,Phellodendri cortex and Gardeniae fructus were 39,93,81 and 88,respectively.After topology analysis,we got 53 core targets from HLJDD potential active ingredients related targets-OSCC related targets PPI network.With the GO enrichment analysis,we found that these core targets enriched in cell cycle,apoptosis,proliferation,mitogen-activated protein kinase(MAPK)activation etc.Additionally,we got TOP30 pathways enrichment from KEGG pathway analysis.Then we treated SCC-25 and CAL-27 cell lines with HLJDD and found that HLJDD could inhibit cell proliferation,induce late apoptosis and inhibit cell invasion and migration.In cell cycle,HLJDD could inhibit G1 phase and arrest in S phase,which was consistent with the network pharmacology results.Finally,we checked MAPK pathway,nuclear factor-kappa B(NF-κB)pathway,protein kinase B(AKT)pathway and so on.It showed that HLJDD might involve in phosphate-extracellular regulated protein kinases1/2(p-ERK1/2),p-NF-κB p65(S468)playing a therapeutic role in OSCC.Conclusion: Our study was the first time using the network pharmacology to explore the mechanism about HLJDD worked on OSCC.The results of network pharmacology provided a new analytical method to screen drug targets for the treatment of OSCC while the results of cell experiments increased the credibility of network pharmacology results.We supposed that HLJDD could be a potential drug for the treatment of OSCC.