title: Suppression of wingless-type MMTV integration site family, member 1 expression by small interfering RNA inhibits U251 glioma cell growthin vitro.
abstract: A Wingless-type MMTV integration site family, member 1 (Wnt-1) RNA interference expression vector was constructed during the present study, which was used to transfect the glioma U251 cell line and investigate its effect on glioma. Two 21-base oligonucleotides complementary to the coding sequence that was flanking the loop sequence were designed to form a DNA hairpin template for the target small interfering RNA (siRNA). The siRNA templates were cloned into the siRNA expression vector, pGPU6/green fluorescent protein (GFP)/Neo and the sequence was confirmed by DNA sequencing. The pGPU6/GFP/Neo-short hairpin RNA (shRNA)-Wnt-1vector was subsequently transfected into U251 cells, and reverse transcription polymerase chain reaction and western blot analysis were used to evaluate theWnt-1gene silencing effect on U251 cell growth by MTT assay and flow cytometry. The Wnt-1 protein expression was significantly reduced following transfection with the recombinant plasmid, as determined by western blot analysis of the transfected U251 cells. This transfection exhibited a significantly higher death rate, as shown by MTT. Thus, the present study demonstrated that the pGPU6/GFP/Neo-shRNA-Wnt-1vector inhibited Wnt-1 protein expression. However, further investigations regarding theWntsignaling pathway in glioma pathogenesis are required.
authors.affiliation: Department of Neurosurgery, Clinical Medical College of Yangzhou University, Yangzhou, Jiangsu 225001, P.R. China.
authors.forename: Lun
authors.initials: L
authors.lastname: Dong
authors.affiliation: Department of Neurosurgery, Clinical Medical College of Yangzhou University, Yangzhou, Jiangsu 225001, P.R. China.
authors.forename: Xiao-Chun
authors.initials: XC
authors.lastname: Duan
authors.affiliation: Central Laboratory, Clinical Medical College of Yangzhou University, Yangzhou, Jiangsu 225001, P.R. China.
authors.forename: Chong-Xu
authors.initials: CX
authors.lastname: Han
authors.affiliation: Department of Neurosurgery, Clinical Medical College of Yangzhou University, Yangzhou, Jiangsu 225001, P.R. China.
authors.forename: Hengzhu
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Neurosurgery, Clinical Medical College of Yangzhou University, Yangzhou, Jiangsu 225001, P.R. China.
authors.forename: Yongkang
authors.initials: Y
authors.lastname: Wu
keywords: RNA interference; U251 cells; glioma; member 1; plasmid; proliferation; wingless-type MMTV integration site family