The cell surface mucin podocalyxin regulates collective breast tumor budding

Breast Cancer Research : BCR
Marcia L GravesCalvin D Roskelley

Abstract

Overexpression of the transmembrane sialomucin podocalyxin, which is known to play a role in lumen formation during polarized epithelial morphogenesis, is an independent indicator of poor prognosis in a number of epithelial cancers, including those that arise in the breast. Therefore, we set out to determine if podocalyxin plays a functional role in breast tumor progression. MCF-7 breast cancer cells, which express little endogenous podocalyxin, were stably transfected with wild type podocalyxin for forced overexpression. 4T1 mammary tumor cells, which express considerable endogenous podocalyxin, were retrovirally transduced with a short hairpin ribonucleic acid (shRNA) targeting podocalyxin for stable knockdown. In vitro, the effects of podocalyxin on collective cellular migration and invasion were assessed in two-dimensional monolayer and three-dimensional basement membrane/collagen gel culture, respectively. In vivo, local invasion was assessed after orthotopic transplantation in immunocompromised mice. Forced overexpression of podocalyxin caused cohesive clusters of epithelial MCF-7 breast tumor cells to bud off from the primary tumor and collectively invade the stroma of the mouse mammary gland in vivo. This budding was no...Continue Reading

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Oct 22, 2016·Current Opinion in Cell Biology·Maren Diepenbruck, Gerhard Christofori
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Jul 3, 2021·Cancers·Ngoc Le TranGuiying Nie

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Methods Mentioned

BETA
fluorescent-activated
FACS
xenograft
xenografts

Software Mentioned

AF6000
Adobe Photoshop
CellSens

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