GSE37472 ( miRNA )

oral cancer ( oral )

Serum miRNA profiles of patients with oral cancer and pre-cancer

Cancer : 50 / Control : 26

Cp values and ROX reference dye normalization were calculated using Viia7 Software (Applied Biosystems). All assays were inspected for distinct melting curves and those assays with more than one Tm and those detected within five Cp’s of the negative control (Cp >35) were excluded from analysis. The miRNAs that gave no signal (null in matrix non-normalized) in any samples were removed before normalization.

The aim of this study was to examine the expression profiles of circulating miRNAs in the serum of patients with high-risk oral lesions (HRLs; oral cancer or carcinoma in situ) and to explore their utility as potential oral cancer biomarkers. Global serum miRNA profiles were generated by quantitative PCR method from 1) patients diagnosed with HRLs and undergoing intent-to-cure surgical treatment (N = 30) and 2) a demographically-matched, normal control group (N = 26).

qPCR miRNA expression profiling of serum samples. The sample size for control and HRL cases are indicated in the summary. The samples were not processed in replicate. Dataset 1: Samples from pre-surgery HRL patients (N = 30) and normal controls (N = 26). Dataset 2: Samples taken both before and after tumor resections (N = 10).

Maclellan SA, Lawson J, Baik J, Guillaud M et al. Differential expression of miRNAs in the serum of patients with high-risk oral lesions. Cancer Med 2012 Oct;1(2):268-74. PMID: 23342275

Total RNA was extracted each from 300uL serum samples using 3D-Gene® RNA extraction reagent from liquid sample kit (Toray Industries, Inc.).

total RNA

SYBR Green

A fixed volume of eluted RNA (19.2 μL for 768 reactions) was used for miRNA expression assays. Total RNA was reverse transcribed using the miRCURY LNA Universal RT miRNA PCR, Polyadenylation and cDNA synthesis kit (Exiqon Inc.). The cDNA was then quantified by quantitative real-time PCR (qRT-PCR) on the miRNA Ready-to-Use PCR, Human panel I and panel II with the miRCURY LNA Universal RT miRNA PCR, SYBR Green master mix according to the manufacturer’s protocol (Exiqon Inc.).

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