title: Sensitive electrochemiluminescence detection of c-Myc mRNA in breast cancer cells on a wireless bipolar electrode.
abstract: We report an ultrasensitive wireless electrochemiluminescence (ECL) protocol for the detection of a nucleic acid target in tumor cells on an indium tin oxide bipolar electrode (BPE) in a poly(dimethylsiloxane) microchannel. The approach is based on the modification of the anodic pole of the BPE with antisense DNA as the recognition element, Ru(bpy)(3)(2+)-conjugated silica nanoparticles (RuSi@Ru(bpy)(3)(2+)) as the signal amplification tag, and reporter DNA as a reference standard. It employs the hybridization-induced changes of RuSi@Ru(bpy)(3)(2+) ECL efficiency for the specific detection of reporter DNA released from tumor cells. Prior to ECL detection, tumor cells are transfected with CdSe@ZnS quantum dot (QD)-antisense DNA/reporter DNA conjugates. Upon the selective binding of antisense DNA probes to intracellular target mRNA, reporter DNA will be released from the QDs, which indicates the amount of the target mRNA. The proof of concept is demonstrated using a proto-oncogene c-Myc mRNA in MCF-7 cells (breast cancer cell line) as a model target. The wireless ECL biosensor exhibited excellent ECL signals which showed a good linear range over 2 × 10(-16) to 1 × 10(-11) M toward the reporter DNA detection and could accurately quantify c-Myc mRNA copy numbers in living cells. C-Myc mRNA in each MCF-7 cell and LO2 cell was estimated to be 2203 and 13 copies, respectively. This wireless ECL strategy provides great promise in a miniaturized device and may facilitate the achievement of point of care testing.
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China.
authors.forename: Mei-Sheng
authors.initials: MS
authors.lastname: Wu
authors.forename: Guang-sheng
authors.initials: GS
authors.lastname: Qian
authors.forename: Jing-Juan
authors.initials: JJ
authors.lastname: Xu
authors.forename: Hong-Yuan
authors.initials: HY
authors.lastname: Chen
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods*; D001943:Breast Neoplasms / Q000473:pathology*; D019187:Cadmium Compounds / Q000737:chemistry; D045744:Cell Line, Tumor; D015342:DNA Probes / Q000737:chemistry; D004563:Electrochemistry; D004566:Electrodes; D006801:Humans; D008163:Luminescent Measurements; D009942:Organometallic Compounds / Q000737:chemistry; D000090063:Proto-Oncogene Mas; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics*; D045663:Quantum Dots; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D018036:Selenium Compounds / Q000737:chemistry; D013440:Sulfides / Q000737:chemistry; D059015:Wireless Technology; D017967:Zinc Compounds / Q000737:chemistry
chemical_list: D019187:Cadmium Compounds; D015342:DNA Probes; C000715470:MAS1 protein, human; D009942:Organometallic Compounds; D000090063:Proto-Oncogene Mas; D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger; D018036:Selenium Compounds; D013440:Sulfides; D017967:Zinc Compounds; C415768:tris(2,2'-bipyridyl)ruthenium(II); C058667:cadmium selenide; C031238:zinc sulfide