Synergistic effects of CD44 and TGF-β1 through AKT/GSK-3β/β-catenin signaling during epithelial-mesenchymal transition in liver cancer cells.
Cancer metastasis is strongly correlated with epithelial-mesenchymal transition (EMT), in which transforming growth factor-β (TGF-β) signaling plays a central role. CD44 has emerged as a cancer stem cell (CSC) marker that strongly induces EMT together with TGF-β1. This study aimed to investigate the link between high CD44 and TGF-β1 levels during EMT in HCC cell lines. FACS analysis showed high expression of CD44 in TGF-β1-positive SNU-368 cells and TGF-β1-negative SNU-354 cells. SNU-368 CD44(+) cells showed EMT through up-regulation of the AKT/GSK-3β/β-catenin pathway. By comparison, SNU-354 CD44(+) cells showed only increased N-cadherin expression, which was not accompanied by a decrease in E-cadherin expression, and also down-regulated the AKT/GSK-3β/β-catenin pathway. However, TGF-β1-stimulated SNU-354 cells (CD44/TGF-β1(+)) exhibited lower E-cadherin and higher N-cadherin expression with increased AKT/GSK-3β/β-catenin pathway activity. CD44/TGF-β1(+) SNU-354 cells also showed enhanced migration and formed larger spheres, while the TGF-β1-induced stem cell properties returned to their original state with the TGF-β1 inhibitor SB431542. SB431542-treated SNU-368 (CD44/TGF-β1(-)) cells also showed diminished N-cadherin and AKT/GSK-3β/β-catenin pathway activity and further decreased cell motility in a wound healing assay. However, CD44 knockdown in SNU-354 cells did not induce EMT even after treatment with TGF-β1. Finally, double inhibition of both CD44 and TGF-β1 further decreased migration and sphere formation more strongly than a single inhibition in SNU-368 cells. In conclusion, the current study demonstrated the synergistic interactions between CD44 and TGF-β1 in EMT induction and CSC properties through the AKT/GSK-3β/β-catenin pathway in HCC cells.
The Catholic University Liver Research Center & WHO Collaborating Center of Viral Hepatitis, The Catholic University of Korea, Seoul, Republic of Korea.
Na Ri
NR
Park
The Catholic University Liver Research Center & WHO Collaborating Center of Viral Hepatitis, The Catholic University of Korea, Seoul, Republic of Korea.
Jung Hoon
JH
Cha
The Catholic University Liver Research Center & WHO Collaborating Center of Viral Hepatitis, The Catholic University of Korea, Seoul, Republic of Korea; Department of Internal Medicine, Seoul St. Mary's Hospital, The Catholic University of Korea, Seoul, Republic of Korea. Electronic address: garden@catholic.ac.kr.
Jeong Won
JW
Jang
The Catholic University Liver Research Center & WHO Collaborating Center of Viral Hepatitis, The Catholic University of Korea, Seoul, Republic of Korea; Department of Internal Medicine, Seoul St. Mary's Hospital, The Catholic University of Korea, Seoul, Republic of Korea. Electronic address: baesh@catholic.ac.kr.
Si Hyun
SH
Bae
The Catholic University Liver Research Center & WHO Collaborating Center of Viral Hepatitis, The Catholic University of Korea, Seoul, Republic of Korea; Department of Internal Medicine, Seoul St. Mary's Hospital, The Catholic University of Korea, Seoul, Republic of Korea.
Bohyun
B
Jang
The Catholic University Liver Research Center & WHO Collaborating Center of Viral Hepatitis, The Catholic University of Korea, Seoul, Republic of Korea.
Jung-Hee
JH
Kim
The Catholic University Liver Research Center & WHO Collaborating Center of Viral Hepatitis, The Catholic University of Korea, Seoul, Republic of Korea.
Wonhee
W
Hur
The Catholic University Liver Research Center & WHO Collaborating Center of Viral Hepatitis, The Catholic University of Korea, Seoul, Republic of Korea; Department of Internal Medicine, Seoul St. Mary's Hospital, The Catholic University of Korea, Seoul, Republic of Korea.
Jong Young
JY
Choi
The Catholic University Liver Research Center & WHO Collaborating Center of Viral Hepatitis, The Catholic University of Korea, Seoul, Republic of Korea; Department of Internal Medicine, Seoul St. Mary's Hospital, The Catholic University of Korea, Seoul, Republic of Korea.
Seung Kew
SK
Yoon
D045744:Cell Line, Tumor; D058750:Epithelial-Mesenchymal Transition; D055785:Gene Knockdown Techniques; D000071679:Glycogen Synthase Kinase 3 beta / Q000378:metabolism*; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics / Q000378:metabolism*; D008113:Liver Neoplasms / Q000201:enzymology / Q000378:metabolism / Q000473:pathology*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D015398:Signal Transduction; D053773:Transforming Growth Factor beta1 / Q000378:metabolism*; D051176:beta Catenin / Q000378:metabolism*
C497483:CD44 protein, human; D018960:Hyaluronan Receptors; D053773:Transforming Growth Factor beta1; D051176:beta Catenin; C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; D051057:Proto-Oncogene Proteins c-akt
CD44; Epithelial-mesenchymal transition (EMT); Hepatocellular carcinoma (HCC); Metastasis; Transforming growth factor-β1 (TGF-β1)