Abstract
Objective To design and verify the PCR primers for SMAD gene family independently , and to explore the feasibility of using these primers for detection of SMAD gene expression in non-small-cell lung cancer ( NSCLC) by RT-PCR.Methods We independently designed and verified SMAD 1-8 specific primers for the SMAD gene family , and iden-tified that the amplification efficiency of SMAD 1-1, SMAD2-1, SMAD3-1, SMAD4, SMAD5-1, SMAD6-1, SMAD7-1, and SMAD8-2 primers was higher and their specificity was stronger.The expression of SMAD1-8 in the normal bronchial epithelial cell line BEAS-2B, lung adenocarcinoma A549, H1299 cells, and lung squamous-cell carcinoma SK-MES-1 cells was detected by RT-PCR.The primers were SMAD1-1, SMAD2-1, SMAD3-1, SMAD4-2, SMAD5-1, SMAD6-1, SMAD7-1, and SMAD8-2, respectively.Results The expression of SMAD1 in the normal bronchial epithelial BEAS-2B cells was taken as the baseline [the revised relative expression (RQ) is 1], the relative expression of SMAD2 was 6.590 ±0.459, SMAD3 was 10.977 ±0.312, SMAD4 was 4.651 ±0.115, SMAD5 was 16.762 ±0.389, SMAD6 was 3.248 ± 0.108, SMAD7 was 2.007 ±0.012, and SMAD8 was 1.144 ±0.103, all were higher than SMAD1 expression, and SMAD5 expression was the highest , followed by SMAD3 expression.The expression of SMAD2, SMAD4, SMAD5, SMAD6, SMAD7 and SMAD8 in the lung adenocarcinoma A549, H1299 cells and squamous-cell lung carcinoma SK-MES-1 cells was lower than that of the normal bronchial epithelial cell line BEAS -2B.The expression of SMAD1 in the H1299 and SK-MES-1 cells was higher than that in the BEAS-2B and A549 cells, and the expression of SMAD1 in the A549 cell was lower than that in the BEAS-2B cells.SMAD3 expression increased in the SK-MES-1 cells, and decreased in the H1299 and A549 cells as compared with that in the BEAS-2B cells.In H1299 cells, the expression of SMAD2, SMAD6 and SMAD7 were lower than that in the other three kinds of cells ; significant difference was found between every two kinds of cells (P＜0.05 or P＜0.01).Conclusions The specific primers for SMAD gene family are successfully de-signed and used to detect the expression of SMAD gene in NSCLC.SMAD5 may be involved in the occurrence and develop-ment of lung cancer , and SMAD3 may be associated with epithelial-related diseases.关键词
非小细胞肺癌/聚合酶链式反应/果蝇母源抗生物皮肤生长因子基因Key words
non-small-cell lung cancer/polymerase chain reaction/SMAD gene分类
医药卫生
