Upregulated SMAD3 promotes epithelial-mesenchymal transition and predicts poor prognosis in pancreatic ductal adenocarcinoma.
In pancreatic ductal adenocarcinoma (PDAC), features of epithelial-mesenchymal transition (EMT) are often seen in tumor tissue, and such features correlate with poor prognosis. Solitary infiltration of tumor cells represents a morphological phenotype of EMT, and we previously reported that a high degree of solitary cell infiltration correlates with EMT-like features, including reduced E-cadherin and elevated vimentin levels. Using solitary cell infiltration to evaluate the degree of EMT, gene-expression profiling of 12 PDAC xenografts was performed, and SMAD3 was identified as an EMT-related gene. Immunohistochemistry using clinical specimens (n=113) showed that SMAD3 accumulated in the nuclei of tumor cells, but was not detected in most epithelial cells in the pancreatic duct. Moreover, SMAD3 upregulation correlated with malignant characteristics, such as higher tumor grade and lymph node metastasis, as well as with EMT-like features. SMAD4, which plays a key role in transforming growth factor-β (TGF-β) signaling, is inactivated in approximately half of PDAC cases. In this study, the nuclear accumulation of SMAD3 was immunohistochemically detected even in SMAD4-negative cases. SMAD3 knockdown resulted in upregulated E-cadherin, downregulated vimentin, and reduced cell motility in pancreatic cancer cells regardless of SMAD4 status. In addition, TGF-β-treatment resulted in EMT induction in cells carrying wild-type SMAD4, and EMT was suppressed by SMAD3 knockdown. Patients with upregulated SMAD3 and a high degree of solitary cell infiltration had shorter times to recurrence and shorter survival times after surgery, and multivariate analysis showed that both factors were independent prognostic factors linked to unfavorable outcomes. These findings suggest that SMAD3 in PDAC is involved in the promotion of malignant potential through EMT induction in tumor cells regardless of SMAD4 status and serves as a potential biomarker of poor prognosis.
Department of Pathology, School of Medicine, Keio University, Tokyo, Japan.
Ken
K
Yamazaki
Department of Pathology, School of Medicine, Keio University, Tokyo, Japan.
Yohei
Y
Masugi
Department of Pathology, School of Medicine, Keio University, Tokyo, Japan.
Kathryn
K
Effendi
Department of Pathology, School of Medicine, Keio University, Tokyo, Japan.
Hanako
H
Tsujikawa
Pathology Division, National Cancer Center Research Institute, Tokyo, Japan.
Nobuyoshi
N
Hiraoka
Department of Surgery, School of Medicine, Keio University, Tokyo, Japan.
Minoru
M
Kitago
Department of Surgery, School of Medicine, Keio University, Tokyo, Japan.
Masahiro
M
Shinoda
Department of Surgery, School of Medicine, Keio University, Tokyo, Japan.
Osamu
O
Itano
Department of Surgery, School of Medicine, Keio University, Tokyo, Japan.
Minoru
M
Tanabe
Department of Surgery, School of Medicine, Keio University, Tokyo, Japan.
Yuko
Y
Kitagawa
Department of Pathology, School of Medicine, Keio University, Tokyo, Japan.
Michiie
M
Sakamoto
D000368:Aged; D021441:Carcinoma, Pancreatic Ductal / Q000235:genetics / Q000378:metabolism* / Q000401:mortality; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism; D016000:Cluster Analysis; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D005260:Female; D055785:Gene Knockdown Techniques; D006801:Humans; D053208:Kaplan-Meier Estimate; D008297:Male; D008875:Middle Aged; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000401:mortality; D011379:Prognosis; D051900:Smad3 Protein / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D015854:Up-Regulation / Q000235:genetics*
D051900:Smad3 Protein