Detection of expression of RRM1 mRNA by real-time fluorescent quantitative PCR in non-small cell lung cancer tissues and peripheral blood
- Author:
Song DONG
1
Author Information
1. Department of Biochip
- Publication Type:Journal Article
- Keywords:
Carcinoma, non-small cell lung;
Gene expression;
Real-time fluorescent quantitative PCR;
Ribonucleotide reductases
- From:
Tumor
2007;27(7):577-581
- CountryChina
- Language:Chinese
-
Abstract:
Objective: We tired to develop a method of detecting the expression level of RRM1 mRNA in tumor tissues and peripheral blood by real-time fluorescent quantitative PCR and compare the detection methods and results between the two different samples. We aimed to provide the basis for predicting clinical outcome from the expression of RRMI gene. Methods: The plasmid standard of RRM1 gene was constructed. Real-time quantitative analysis was performed using ABI7000 PCR kit. The PCR condition was optimized and the standard curve was established to detect the clinical samples. Results: The RRMI gene expression was detected in lung cancer tissues and normal lung tissues from 18 patients and in peripheral blood samples from 17 patients. The relative expression of RRM1 normalized by β-actin was 4.46 × 10-3 in lung cancer tissues, 3.43 × 10-3 in normal tissues, 2.54 × 10-3 in peripheral blood. The linear correlation between Ct value and the logarithm of original concentration was favorable. Conclusion: The expression of RRM1 mRNA in non-small cell lung cancer tissues and peripheral blood was successfully detected by using SYBR Green I fluorescence real-time quantitative PCR assay. There is no significant difference between the two sources of samples. The detection method has relatively higher sensitivity and specificity and can be used in clinical analysis.
