Construction and identification of small interfering RNA targeting silence Trop2 expression in human laryngeal carcinoma cell line HEP2

Objective To construct the small interfering RNA( si RNA) that can silence Trop2 expression and to investigate the silencing effect of si RNA on Trop2 gene in human laryngeal carcinoma cell line HEP2. Methods Three different gene sequences si RNA specific targeting Trop2 gene were constructed and used to transfect into HEP2 cells by using LipofectamineTM2000. The negative control group and blank control group were set up at the same time. These cells were divided into the W group( un-transfection),the NC group( transfection with negative control),the T1 group( S1sequence),the T2 group( S2 sequence) and the T3 group( S3 sequence). The transfection efficiency was reported by green fluorescence protein expression and gene inhibition efficiency was analyzed by real-time RT-PCR. Results Green fluorescence observed by fluorescence microscopy showed that si RNA had been transfected into HEP2 cells,and the transfection ratio was 90%. The expression levels of Trop2 m RNA in cells transfected with Trop2-specific si RNA were lower than those in the untransfected cells or cells transfected with negative control si RNA. Compared with the T2 and T3 groups,si RNA in the T1 group had the strongest inhibitory effect on the expression of Trop2 m RNA,and the difference was significant( 0. 470 ± 0. 063 vs 0. 749 ± 0. 024,0. 824 ± 0. 027,P < 0. 05). Conclusion Effective si RNA sequence has been successfully constructed and selected which can inhibit the expression of Trop2 gene in human laryngeal carcinoma cell line HEP2. It is fundamental for the study on the function of Trop2 gene and the gene therapy of laryngeal carcinoma.