Effect of Twist1 gene on migration and invasion of esophageal squamous cell carcinoma / 中国医师进修杂志

ABSTRACT

Objective To investigate the expression and clinicopathological significance of Twist1 gene in esophageal squamous cell carcinoma(ESCC),and its effect on migration and invasion of ESCC.Methods The ESCC tissues and adjacent normal tissues of 30 ESCC patients having underwent surgery were selected. The expression levels of Twist1 mRNA and protein were detected by real time fluorescence quantitative polymerase chain reaction (RT- qPCR) and Western blot methods. The expression level of Twist1 mRNA in adjacent normal tissues was used as control,the relative expression levels of Twist1 mRNA in 3 ESCC cell lines K70,K140 and EC109 were detected,and the relationship between the expression level and clinicopathological features was analyzed. ESCC cell line EC109 was selected for cell scratch test and Transwell chamber invasion assay respectively.Twist1 small interfering RNA and control double stranded nonsense RNA were transfected respectively, and the changes were compared.Results The expression level of Twist1 mRNA in ESCC tissue was significantly higher than that in adjacent normal tissue(1.91 ± 0.93 vs.0.54 ± 0.26),and there was statistical difference(P<0.01). The relative expression levels of Twist1 mRNA in 3 ESCC cell lines K70, K140 and EC109 were significantly higher than those in adjacent normal tissue (2.30 ± 0.34, 1.78 ± 0.28 and 4.37 ± 0.25 vs. 1.00 ± 0.48), and there were statistical differences (P<0.05). The expression level of Twist1 protein in ESCC tissue was significantly higher than that in adjacent normal tissue (1.35 ± 0.66 vs. 0.25 ± 0.05), and there was statistical difference (P<0.05). The relative expression level of Twist1 mRNA in ESCC tissue had correlation with lymph node metastasis and invasion degree(P<0.05).The expression levels of Twist1 mRNA in ESCC cell lines EC109 after transfected Twist1 small interfering RNA and double stranded nonsense RNA were 0.45 ± 0.16 and 1.00 ± 0.20, the express levels of Twist1 protein were 0.24 ± 0.09 and 0.58 ± 0.10,and there were statistical differences(P<0.05).The cell scratch test result showed that the migration rates of 36 and 72 h of ESCC cell line EC109 were significantly lower than those of the control double stranded nonsense RNA,after Twist1 expression was down regulated by RNA interference(32.6 ± 4.7)% vs.(16.2 ± 6.0)% and(71.9 ± 4.7)% vs.(53.2 ± 6.6)%,there were statistical differences(P<0.05).The Transwell chamber invasion assay result showed that the total number of cells in the bottom of Twist1 through the small interfering RNA of EC109 was lower than that of the transfected control double stranded nonsense RNA (92.5 ± 8.1 vs. 32.7 ± 7.5), and there was statistical difference (P<0.05). Conclusions Twist1 gene may be highly abnormally expressed in ESCC, which has some correlation with invasion and lymph node metastasis,and may have some influences on the migration and invasion of ESCC.