Exogenous Cripto-1 Suppresses Self-Renewal of Cancer Stem Cell Model.
Cripto-1 is a glycophosphatidylinositol (GPI) anchored signaling protein of epidermal growth factor (EGF)-Cripto-1-FRL1-Cryptic (CFC) family and plays a significant role in the early developmental stages and in the different types of cancer cells, epithelial to mesenchymal transition and tumor angiogenesis. Previously, we have developed cancer stem cells (miPS-LLCcm) from mouse iPSCs by culturing them in the presence of conditioned medium of Lewis Lung Carcinoma (LLC) cells for four weeks. Nodal and Cripto-1 were confirmed to be expressed in miPS-LLCcm cells by quantitative reverse transcription PCR (rt-qPCR) implying that Cr-1 was required in maintaining stemness. To investigate the biological effect of adding exogenous soluble CR-1 to the cancer stem cells, we have prepared a C-terminally truncated soluble form of recombinant human CR-1 protein (rhsfCR-1), in which the GPI anchored moiety was removed by substitution of a stop codon through site-directed mutagenesis. rhsfCR-1 effectively suppressed the proliferation and sphere forming ability of miPS-LLCcm cells in a dose-dependent manner in the range of 0 to 5 µg/mL, due to the suppression of Nodal-Cripto-1/ALK4/Smad2 signaling pathway. Frequency of sphere-forming cells was dropped from 1/40 to 1/69 by rhsfCR-1 at 1 µg/mL. Moreover, rhsfCR-1 in the range of 0 to 1 µg/mL also limited the differentiation of miPS-LLCcm cells into vascular endothelial cells probably due to the suppression of self-renewal, which should reduce the number of cells with stemness property. As demonstrated by a soluble form of exogenous Cripto-1 in this study, the efficient blockade would be an attractive way to study Cripto-1 dependent cancer stem cell properties for therapeutic application.
['Alam|Md Jahangir|MJ|0000-0003-1587-0765', 'Takahashi|Ryota|R|', 'Afify|Said M|SM|0000-0003-4026-1400', 'Oo|Aung Ko Ko|AKK|0000-0002-2664-7115', 'Kumon|Kazuki|K|', 'Nawara|Hend M|HM|', 'Khayrani|Aprilliana Cahya|AC|0000-0001-9160-7446', 'Du|Juan|J|', 'Zahra|Maram H|MH|', 'Seno|Akimasa|A|0000-0002-9664-9490', 'Salomon|David S|DS|', 'Seno|Masaharu|M|0000-0001-8547-6259']
D000818:Animals D002454:Cell Differentiation D002460:Cell Line D000066673:Cell Self Renewal D058851:GPI-Linked Proteins D006801:Humans D036341:Intercellular Signaling Peptides and Proteins D051379:Mice D009363:Neoplasm Proteins D009369:Neoplasms D014411:Neoplastic Stem Cells D011994:Recombinant Proteins D015398:Signal Transduction D051899:Smad2 Protein
Cripto-1 self-renewal cancer stem cells miPS-LLCcm mouse iPS recombinant Cripto-1