title: A microchip for integrated single-cell genotoxicity assay.
abstract: With the development of large-scale biologic databases, precision medicine is becoming a frontier in biomedical research. As a main focus of precision medicine study, cancer has been widely accepted as a disease born out of inherited genetic variations or accumulating genomic damage. At the single-cell level, microfluidics or lab-on-a-chip technology for cancer study is an emerging tool for improving risk assessment, diagnostic categories and therapeutic strategies. This work presents a multi-layer microchip for single-cell gene expression profiling. Treated by three drug reagents (i.e. methyl methanesulfonate, docetaxel and colchicine) with varied concentrations and time lengths, individual human breast cancer cells (MCF-7) are then lysed on-chip, and the released mRNA templates are captured and reversely transcribed into cDNA on microbead surface. Three genes (GAPDH, CDKN1A, AURKA) are amplified and quantified simultaneously through triplex real-time polymerase chain reactions (qPCR). Readout per run is set to be eighteen, and can be further improved following same approach. The microchip is able to integrate all steps of single-cell gene expression profiling, and provide precision study of drug induced genotoxicity with reduced reagents consumption per reaction and instrumental cost.
authors: School of Mechanical Engineering and Automation, Fuzhou University, Fujian 350116, China.
authors: Hui
authors: H
authors: Dong
authors: School of Mechanical Engineering and Automation, Fuzhou University, Fujian 350116, China. Electronic address: sh@fzu.edu.cn.
authors: Hao
authors: H
authors: Sun
authors: Department of Medical Oncology, Fujian Provincial Hospital, Fujian 350001, China.
authors: Jianping
authors: J
authors: Zheng
mesh_terms: D064096:Aurora Kinase A / Q000235:genetics*; D003078:Colchicine / Q000494:pharmacology / Q000633:toxicity*; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics*; D000077143:Docetaxel; D020869:Gene Expression Profiling; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics*; D006801:Humans; D056656:Lab-On-A-Chip Devices; D061986:MCF-7 Cells; D008741:Methyl Methanesulfonate / Q000494:pharmacology / Q000633:toxicity*; D044085:Microfluidics; D009152:Mutagenicity Tests; D060888:Real-Time Polymerase Chain Reaction; D043823:Taxoids / Q000494:pharmacology / Q000633:toxicity*
chemical_list: C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D043823:Taxoids; D000077143:Docetaxel; D008741:Methyl Methanesulfonate; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; C576917:AURKA protein, human; D064096:Aurora Kinase A; D003078:Colchicine
keywords: Genotoxicity assay; Integrated microfluidics; Single-cell multiplex qPCR