Effect of Wnt-1 antisense RNA on the outgrowth of a mammary adenocarcinoma cell line expressing that oncogene

Polanec, Janja; Pavelic, Zlatko P.; Myers, W. L.

doi:10.1136/mp.49.3.m166

Cited by 2 publications

(1 citation statement)

References 8 publications

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“…However, few studies have used Wnt-1 gene RNAi. Certain studies have used fragments of chemically synthesized siRNA in transfected cells ( 25 , 26 ), or plasmids lacking a reporter gene ( 27 ), which complicates the assessment of transfection efficiency and the identification of stable cell lines. A previous study described the transfection of siRNA targeting Wnt-1 into SH-SY5Y neuroblastoma cells using Lipofectamine TM 2000 ( 28 ).…”

Section: Discussionmentioning

confidence: 99%

Suppression of wingless-type MMTV integration site family, member 1 expression by small interfering RNA inhibits U251 glioma cell growth in vitro

et al. 2014

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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-1 vector was subsequently transfected into U251 cells, and reverse transcription polymerase chain reaction and western blot analysis were used to evaluate the Wnt-1 gene 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-1 vector inhibited Wnt-1 protein expression. However, further investigations regarding the Wnt signaling pathway in glioma pathogenesis are required.

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