Construction of PLAGL2 siRNA Expression DNA Plasmid and the Interfering Effect on Gene Expression in H441 Cells / 华中科技大学学报(医学版)

ABSTRACT

Objective To construct human PLAGL2 siRNA expression DNA plasmid and study the interfering effect on gene expression in H441 cells. Methods H441 cells were transfected with PLAGL2 siRNA expression DNA plasmid with Fu-gene6. Real time PCR and Western blot were employed to detect the interfering effect on the expression of PLAGL2.SP-CSP-B mRNA and protein in wildtype and PLAGL2 siRNA expression DNA plasmid transfected H441 cells respectively. Results Real time PCR revealed that,as compared with wildtype H441 cells, the expression level of PLAGL2 mRNA in PLAGL2 siRNA plasmid transfected H441 cells was significantly reduced(P<0. 05) ,but that of SP-C mRNA in PLAGL2 siRNA plasmid transfected H441 cells was significantly increased(P<0. 05). Western blot showed that,as compared with wildtype H441 cells, the expression level of PLAGL2 protein in PLAGL2 siRNA DNA plasmid transfected H441 cells was significantly reduced(P< 0. 05). Conclusion Human PLAGL2 siRNA expression DNA plasmid was constructed successfully,and its transfection into the H441 cells could effectively inhibit the PLAGL2 expression,and simultaneously promote the expression of SP-C.