Abstract: OBJECTIVE To develop a more simple, efficient and economical method of total RNA extracted from human non-small cell lung cancer tissues by optimization of traditional TRIzol method, and detect mRNA expression levels of ERCC1, RRM1 and BRCA1. METHODS Thirty-six samples of non-small cell lung cancer(NSCLC, including tumor tissues and normal tissues) were collected and RNA from these samples were extracted through traditional and optimized method, expression levels of ERCC1, RRM1, BRCA1 and housekeeping gene GAPDH SYBR were dectected by RT-PCR. RESULTS The concentration and purity of RNA extracted by traditional method were (0.76±0.07)mg·mL^-1 and 1.87±0.04, respectively, the concentration and purity of RNA extracted by the optimized method performed better, which were (1.12±0.07)mg·mL^-1 and 1.95±0.03, respectively. The agarose gel electrophoresis showed no degradation of RNA, each primer melting curve showed a single peak and good specificity. According to the fluorescence-based real-time detection, ERCC1, RRM1 and BRCA1 mRNA levels in tumor tissues were respectively higher than those of normal tissues. No significance in expression of ERCC1 and BRCA1 between adenocarcinoma and squamous carcinoma was found. Nevertheless, RRM1 expression level in lung squamous carcinoma was significantly higher than that of adenocarcinoma(P<0.05). In addition, gene expression related to gender had no significant difference. CONCLUSION The concentration, purity and integrity of extracted total RNA meet the needs of a variety of gene expression and quantitative detection experiments. The expression levels of ERCC1, RRM1 and BRCA1 in NSCLC is closely correlated with tumor proliferative activity and clinical efficacy of adjuvant chemotherapy for cancer.