Membrane-bound full-length Sonic Hedgehog identifies cancer stem cells in human non-small cell lung cancer
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Etienne Giroux Leprieur1,2,3,*, Bhairavi Tolani1,*, Hui Li1,*, Fleur Leguay1, Ngoc T. Hoang1, Luis A. Acevedo1, Joy Q. Jin1, Hsin-Hui Tseng1, Dongsheng Yue1,4, Il-Jin Kim1, Marie Wislez5, Changli Wang4, David M. Jablons1 and Biao He1
1Thoracic Oncology Program, Department of Surgery, Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, CA, USA
2Department of Respiratory Diseases and Thoracic Oncology, APHP - Ambroise Pare Hospital, Boulogne-Billancourt, France
3EA 4340, UVSQ, Paris-Saclay University, Boulogne-Billancourt, France
4Department of Lung Cancer, Lung Cancer Center, Tianjin Medical University Cancer Institute and Hospital, Tianjin, China
5Sorbonne University, UPMC GRC-04 Theranoscan, Department of Respiratory Diseases, APHP - Tenon Hospital, Paris, France
*These authors have contributed equally to this work
Biao He, email: [email protected]
Keywords: non-small cell lung cancer (NSCLC), Sonic Hedgehog (Shh), cancer stem cells (CSCs), GDC0449, chemo-resistance
Received: July 23, 2017 Accepted: September 05, 2017 Published: October 10, 2017
The mechanism of Sonic Hedgehog (Shh) pathway activation in non-small cell lung cancer (NSCLC) is poorly described. Using an antibody against the Shh C-terminal domain, we found a small population of Shh-positive (Shh+) cells in NSCLC cells. The objective of this study was to characterize these Shh+ cells. Shh+ and Shh- cells were sorted by using Fluorescence Activated Cell Sorting (FACS) on 12 commercial NSCLC cell lines. Functional analyses on sorted cells were performed with gene expression assays (qRT-PCR and microarray) and cells were treated with cytotoxic chemotherapy and a targeted inhibitor of Shh signaling (GDC0449). We used in vivo models of nude mice inoculated with Shh+ and Shh- sorted cells and drug-treated cells. Finally, we confirmed our results in fresh human NSCLC samples (n=48) paired with normal lung tissue. We found that Shh+ cells produced an uncleaved, full-length Shh protein detected on the membranes of these cells. Shh+ cells exerted a paracrine effect on Shh- cells, inducing their proliferation and migration. Shh+ cells were chemo-resistant and showed features of cancer stem cells (CSCs) in vitro and in vivo. Pharmacological inhibition of the Shh pathway suppressed their CSC features. A high percentage of Shh+ cells was associated with poor prognosis in early-stage NSCLC patients. In conclusion, we describe for the first time the presence of an abnormal membrane-bound full-length Shh protein in human cancer cells that allows the identification of CSCs in vitro and in vivo.
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