Real time quantitative PCR analysis of serum circulating miRNAs from patients with head and neck tumors. [exp2]

The samples were drawn from patients with diagnosed oropharyngeal cancer. We used blood samples (10ml) collected from patients after diagnosis, before treatment. Blood was centrifuged for 10 min, after which serum was aspirated, aliquoted in RNAse-free tubes, stored at −80°C until required for further processing and did not undergo more than two freeze-thaw cycles. MicroRNA was extracted from 200 μL serum using miRCURY RNA Isolation kit (Qiagen) following the manufacturer’s protocol and stored at −80°C. miRNA qPCR assays from 70 patients with HnN cancer before irradiation. Equal amount of RNA was used from each patient.