title: Reverse-transcriptase polymerase chain reaction to detect extracellular mRNAs.
abstract: The presence of extracellular nucleic acids has been reported in serum/plasma from cancer and diabetes patients that may help in disease diagnosis. Taking insulin-producing cells as examples here, RT-PCR was used to investigate a correlation between the presence and amounts of extracellular mRNA(s) and cell mass and/or function. RT-PCR was performed on a range of mRNAs, including Pdx1, Npy, Egr1, Pld1, Chgb, InsI, InsII, and Actb in biological triplicate analyses.Reproducible amplification of these mRNAs from MIN6, MIN6 B1, and Vero-PPI cells and their CM suggests that beta cells transcribe and release these mRNAs into their environment. mRNAs secreted from insulin-producing cells into their extracellular environment may have potential as extracellular biomarkers for assessing beta cell mass and function.
authors: School of Pharmacy & Pharmaceutical Sciences, Panoz Institute, Trinity College Dublin, Ireland. srani@tcd.ie
authors: Sweta
authors: S
authors: Rani
authors: Lorraine
authors: L
authors: O'Driscoll
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000032:analysis*; D002522:Chlorocebus aethiops; D017077:Culture Media, Conditioned; D005110:Extracellular Space / Q000378:metabolism*; D020869:Gene Expression Profiling; D006801:Humans; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000378:metabolism; D009369:Neoplasms / Q000378:metabolism; D009360:Neoplastic Cells, Circulating / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014709:Vero Cells
chemical_list: D014408:Biomarkers, Tumor; D017077:Culture Media, Conditioned; D012333:RNA, Messenger