EMT and Dissemination PrecedePancreatic Tumor Formation
Department of Biochemistry and Molecular Biology
Reporter: Ma Ting (马婷 )
DOI10.1016/j.cell.2011.11.025
Metastasis: the final step in a progressive” Darwinian” sequence?
Two clinical observation:The appearance of metastatic lesions years
after resection of small tumors with no clinically evident metastases at diagnosis
metastases of unknown primary, account for as many as 4–5% of all clinical metastases
Metastasis : an inherent feature of a tumor very early in its natural history?
How to identify markers that distinguish cancer cells from normally residing cells in the bloodstream or at the sites of seeding?
How to study EMT in vivo?
A lineage labeling system
• Mouse models : PKCY micePdx1-Cre--Kras and P53 mutations--RosaYFP • Mouse models : IKCY micePdx1-Cre--Kras mutations--p16Ink4a/Arf deletion--RosaYFP • Control mouse models: CY micePdx1-Cre; RosaYFP (CY) animals
• Pdx1: only active in endoderm-derived pancreatic cells• P53、 Kras: mutated with high frequency in human pancreatic cancers• Rosa YFP: track pancreatic epithelial cells during tumor progression, resulting in highly specific and efficient
(>95%) labeling
Enhanced Detection of EMT using Epithelial Lineage Tracing
Cells in the intermediate stage of EMT :Cells co-expressed an epithelial marker(E-cad) and a mesenchymal marker (zeb1/Fsp1) -----10% O
Cells completed an EMT :Cells co-expressed an epithelial marker(E-cad) and a mesenchymal marker (zeb1/Fsp1)-----42% P、 Q
EMT was not detected inlineage-labeled CY control mice by either method L-N
EMT in Premalignant Lesions
EMT was also prevalent in areas of acinar-to-ductal metaplasia (ADM), particularly in ADMIs (acinar-to-ductal metaplasia with Inflammation)
”PanIN mice” :8- to 10-week-old PKCY ,only precancerous PanIN lesions and no histological evidence of carcinoma
EMT occurs in PanIN lesions and ADMIs prior to tumor formation.
Pancreatic Epithelial Cells Spread before Tumor Formation
Individual YFP+ cells :“delamination” in PanIN 2 and 3 lesions. Most of these cells expressed zeb1 and had acquired a fibroblast-like morphology, making them indistinguishable from surrounding stromal cells by conventional histology
YFP+ circulating pancreatic cells (CPCs) were readily detected in the blood by flowcytometry in PDAC mice n E
CPCs were also abundant in the bloodstream of PKCY PanIN C-G
carry the Gly→Asp mutation at codon 12 of the Kras cDNA
Cells derived from the pancreatic epithelium are present in the circulation of mice
with no evidence of carcinoma
Pancreatic Epithelial Cells Spread before Tumor Formation
• CPCs from PanIN mice might seed distant organs?
16/20 of PDAC mice have liver/lung metastasis 0 metastasis in PanIN mice but 4/11YFP+ cells seed in liver (Most were single cells lacated near blood vessels ,without zeb1 or E-cad)0 YFP+ cells seed in liver in CY control mice
Characterization of CPCs
The vast majority of CPCs do not survive passage through the pulmonary circulation
There were no statistically significant differences in cell-surface phenotype between CPCs from the two groups
CPCs from PDAC and PanIN animals are phenotypically similar and that a large fraction maintains a mesenchymal phenotype in the circulation.
CPCs Exhibit Features of Cancer Stem Cells
In human pancreatic tumors, cancer stem cells may be contained within a CD24+CD44+ population of cells
CPCs might exhibit features of CSCs
Cells that Have Undergone EMT Have Tumor-Initiating Properties
Inflammation Promotes EMT, Invasiveness, and Dissemination
Highlights of this research
• Invading cells exhibit EMT in an autochthonous model of pancreatic cancer• Mutant cells enter the circulation before cancer is found on histology• Circulating pancreatic cells (CPCs) express cancer stem cell-associated markers• Inflammation is necessary and sufficient for EMT, invasion, and dissemination