PMID- 26923077 OWN - NLM STAT- MEDLINE DCOM- 20161213 LR - 20161230 IS - 1502-7686 (Electronic) IS - 0036-5513 (Linking) VI - 76 IP - 3 DP - 2016 TI - Ultra-micro samples can be used for mRNA quantification of lung cancer biomarkers. PG - 243-8 LID - 10.3109/00365513.2016.1143114 [doi] AB - BACKGROUND: Isolating sufficient material for molecular testing remains challenging in non-small cell lung cancer (NSCLC). The use of new ultra-microsamples (uMS) is proven sufficient for DNA and mRNA detection, but whether uMS are useful for quantifying mRNA expression is unknown. We investigated if uMS from lung cancer patients can be used to generate quantitative data on mRNA expression. METHODS: uMS were collected from primary tumors and lymph nodes from patients suspected of having lung cancer. mRNA was isolated, reverse-transcribed into cDNA and quantified with quantitative PCR assays for hepatocyte growth factor receptor (MET), hepatocyte growth factor (HGF), epidermal growth factor receptor (EGFR) and amphiregulin (AREG) mRNA. The fraction of tumor cells to normal cells was estimated in each sample. RESULTS: MET, HGF, EGFR, and AREG expression were evaluated in 90 samples (30 containing cancer cells and 60 without cancer cells). MET and EGFR expression were negligible in samples without cancer cells. In samples containing cancer cells, MET and EGFR could be quantified in 13 samples each. Adjustment for tumor-cell fraction made it possible to obtain a quantitative result for the tumor-cell mRNA expression of MET and EGFR. In contrast, AREG and HGF were expressed in samples without tumor cells. These samples were used to establish the AREG and HGF mRNA expression in normal cells. Seven out of 14 AR-positive and two out of eight HGF-positive samples with tumor cells were above a cut-off of the mean + 2SD established in samples without tumor cells. CONCLUSION: We demonstrate that uMS contain high-quality mRNA, and quantitative studies can be performed when the tumor-cell fraction is considered. FAU - Sandfeld-Paulsen, Birgitte AU - Sandfeld-Paulsen B AD - a Department of Oncology , Aarhus University Hospital , Denmark ; AD - b Department of Clinical Biochemistry , Aarhus University Hospital , Denmark ; FAU - Demuth, Christina AU - Demuth C AD - b Department of Clinical Biochemistry , Aarhus University Hospital , Denmark ; FAU - Folkersen, Birgitte H AU - Folkersen BH AD - c Department of Pulmonary Medicine , Aarhus University Hospital , Denmark ; FAU - Rasmussen, Torben R AU - Rasmussen TR AD - c Department of Pulmonary Medicine , Aarhus University Hospital , Denmark ; FAU - Madsen, Line B AU - Madsen LB AD - d Department of Pathology , Aarhus University Hospital , Denmark. FAU - Sorensen, Boe S AU - Sorensen BS AD - b Department of Clinical Biochemistry , Aarhus University Hospital , Denmark ; FAU - Meldgaard, Peter AU - Meldgaard P AD - a Department of Oncology , Aarhus University Hospital , Denmark ; LA - eng PT - Journal Article DEP - 20160229 PL - England TA - Scand J Clin Lab Invest JT - Scandinavian journal of clinical and laboratory investigation JID - 0404375 RN - 0 (Biomarkers, Tumor) RN - 0 (RNA, Messenger) SB - IM MH - Biomarkers, Tumor/genetics/isolation & purification/*metabolism MH - Carcinoma, Non-Small-Cell Lung/*diagnosis/metabolism MH - Case-Control Studies MH - Humans MH - Lung/metabolism MH - Lung Neoplasms/*diagnosis/metabolism MH - RNA, Messenger/genetics/isolation & purification/*metabolism MH - Real-Time Polymerase Chain Reaction MH - Reverse Transcriptase Polymerase Chain Reaction OTO - NOTNLM OT - EBUS-TBNA OT - EUS-FNA OT - FNA OT - NSCLC OT - SCLC OT - mRNA EDAT- 2016/03/01 06:00 MHDA- 2016/12/15 06:00 CRDT- 2016/03/01 06:00 PHST- 2016/03/01 06:00 [entrez] PHST- 2016/03/01 06:00 [pubmed] PHST- 2016/12/15 06:00 [medline] AID - 10.3109/00365513.2016.1143114 [doi] PST - ppublish SO - Scand J Clin Lab Invest. 2016;76(3):243-8. doi: 10.3109/00365513.2016.1143114. Epub 2016 Feb 29.