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Wnt2 knock down by RNAi inhibits the proliferation of in vitro-cultured human keloid fibroblasts.


ABSTRACT: To study the effect of knocking down wingless-related MMTV integration site 2 (Wnt2) expression by RNAi on the growth and signaling pathways of ex vitro-cultured keloid fibroblasts (KFB).Human KFB were isolated from 10 keloid patient specimens. The KFB cells were then transfected with 4 pairs of small interfering RNA (siRNA) targeting human Wnt2, respectively. Reverse transcriptase-polymerase chain reaction and Western blot analysis were conducted to verify the knock down of Wnt2, and the expression of ?-catenin glycogen synthase kinase-3? (GSK-3?) and cyclin D1 were examined.siRNA Wnt2 transfection (siWnt2) resulted in the significant inhibition of Wnt2 expression at both the mRNA and protein levels. The expression of ?-catenin, GSK-3?, p-GSK-3?, and cyclin D1 at the protein level also decreased in siWnt2 cells. siWnt2 resulted in a substantially slower growth and significant delay in cell doubling time of the KFB cells compared with control groups. Further, the siRNA knock down of GSK-3? and ?-catenin resulted in slower proliferation rates, respectively.Wnt2 siRNA has an inhibitive effect on keloid fibroblast proliferation, which may be a potential therapeutic approach for keloid and other human fibrotic diseases.

SUBMITTER: Cai Y 

PROVIDER: S-EPMC6156062 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature

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Wnt2 knock down by RNAi inhibits the proliferation of in vitro-cultured human keloid fibroblasts.

Cai Yumei Y   Yang Weiqun W   Pan Mingmeng M   Wang Chaoyang C   Wu Wenyi W   Zhu Shize S  

Medicine 20180901 37


To study the effect of knocking down wingless-related MMTV integration site 2 (Wnt2) expression by RNAi on the growth and signaling pathways of ex vitro-cultured keloid fibroblasts (KFB).Human KFB were isolated from 10 keloid patient specimens. The KFB cells were then transfected with 4 pairs of small interfering RNA (siRNA) targeting human Wnt2, respectively. Reverse transcriptase-polymerase chain reaction and Western blot analysis were conducted to verify the knock down of Wnt2, and the expres  ...[more]

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