Paweena Dana1,2,3, Saowaluk Saisomboon1,2, Ryusho Kariya3, Seiji Okada3, Sumalee Obchoei4, Kanlayanee Sawanyawisuth1,2, Chaisiri Wongkham1,2, Chawalit Pairojkul2,5, Sopit Wongkham1,2,3, Kulthida Vaeteewoottacharn6,7,8. 1. Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40005, Thailand. 2. Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, 40002, Thailand. 3. Division of Hematopoiesis, Joint Research Center for Human Retrovirus Infection and Graduate School of Medical Sciences, Kumamoto University, Kumamoto, 860-0811, Japan. 4. Department of Biochemistry, Faculty of Science, Prince of Songkla University, Songkhla, 90110, Thailand. 5. Department of Pathology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand. 6. Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40005, Thailand. kulthidava@kku.ac.th. 7. Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, 40002, Thailand. kulthidava@kku.ac.th. 8. Division of Hematopoiesis, Joint Research Center for Human Retrovirus Infection and Graduate School of Medical Sciences, Kumamoto University, Kumamoto, 860-0811, Japan. kulthidava@kku.ac.th.
Abstract
PURPOSE: Cholangiocarcinoma (CCA) is an aggressive type of cancer. The major obstacles for treatment are its late presentation and the occurrence metastases. Targeting the metastatic process may serve as a treatment option. CD147 is a membrane protein that promotes CCA metastasis. High lactate levels in CCA are predicted to result from lactate dehydrogenase A expression and sensitivity to monocarboxylate transporter (MCT) inhibitors. An involvement of CD147 in MCT maturation has been reported, but the exact role of MCT in CCA is not clear. Here, we aimed to assess the mechanism of CD147-promoted CCA progression through MCT regulation. METHODS: The expression levels of CD147 and MCT-1/4 in human CCA tissues were determined by immunohistochemistry. Two CD147 knockout (CD147 KO) CCA cell (KKU-213) clones were established using the CRISPR/Cas9 system. Cell migration and invasion were determined using a Boyden chamber assay. Temporal protein levels were modified by siRNA, specific inhibitors and/or activators. The expression of target proteins was determined using Western blot analyses. RESULTS: CD147 and MCT-1/4 were found to be overexpressed in CCA tissues compared to normal bile duct tissues. In addition, we found that CD147 knockdown significantly alleviated CCA cell migration and invasion, concomitant with decreased pAkt, pFoxO3, pNF-κB (pp65) and MCT-1/4 levels. Conversely, we found that FoxO3 knockdown led to recovered migration/invasion abilities and increased pp65 and MCT-1/4 expression levels. The involvement of Akt in the regulation of MCT-1/4 expression through CD147 was established by inhibition and activation of Akt phosphorylation. CONCLUSION: Our data indicate that CD147 promotes the malignant progression of CCA cells by activating the Akt-FoxO3-NF-κB-MCT-1/4 axis. As such, CD147 may serve as a possible target for advanced CCA treatment.
PURPOSE:Cholangiocarcinoma (CCA) is an aggressive type of cancer. The major obstacles for treatment are its late presentation and the occurrence metastases. Targeting the metastatic process may serve as a treatment option. CD147 is a membrane protein that promotes CCA metastasis. High lactate levels in CCA are predicted to result from lactate dehydrogenase A expression and sensitivity to monocarboxylate transporter (MCT) inhibitors. An involvement of CD147 in MCT maturation has been reported, but the exact role of MCT in CCA is not clear. Here, we aimed to assess the mechanism of CD147-promoted CCA progression through MCT regulation. METHODS: The expression levels of CD147 and MCT-1/4 in humanCCA tissues were determined by immunohistochemistry. Two CD147 knockout (CD147 KO) CCA cell (KKU-213) clones were established using the CRISPR/Cas9 system. Cell migration and invasion were determined using a Boyden chamber assay. Temporal protein levels were modified by siRNA, specific inhibitors and/or activators. The expression of target proteins was determined using Western blot analyses. RESULTS:CD147 and MCT-1/4 were found to be overexpressed in CCA tissues compared to normal bile duct tissues. In addition, we found that CD147 knockdown significantly alleviated CCA cell migration and invasion, concomitant with decreased pAkt, pFoxO3, pNF-κB (pp65) and MCT-1/4 levels. Conversely, we found that FoxO3 knockdown led to recovered migration/invasion abilities and increased pp65 and MCT-1/4 expression levels. The involvement of Akt in the regulation of MCT-1/4 expression through CD147 was established by inhibition and activation of Akt phosphorylation. CONCLUSION: Our data indicate that CD147 promotes the malignant progression of CCA cells by activating the Akt-FoxO3-NF-κB-MCT-1/4 axis. As such, CD147 may serve as a possible target for advanced CCA treatment.
Authors: Hakryul Jo; Subhanjan Mondal; Dewar Tan; Eiichiro Nagata; Shunya Takizawa; Alok K Sharma; Qingming Hou; Kumaran Shanmugasundaram; Amit Prasad; Joe K Tung; Alexander O Tejeda; Hengye Man; Alan C Rigby; Hongbo R Luo Journal: Proc Natl Acad Sci U S A Date: 2012-06-11 Impact factor: 11.205
Authors: Min Tang; Yan Zhao; Nanjing Liu; E Chen; Zhen Quan; Xiaohou Wu; Chunli Luo Journal: J Cancer Res Clin Oncol Date: 2017-02-22 Impact factor: 4.553
Authors: Noriaki Tanaka; Mei Zhao; Lin Tang; Ameeta A Patel; Qing Xi; Hieu T Van; Hideaki Takahashi; Abdullah A Osman; Jiexin Zhang; Jing Wang; Jeffrey N Myers; Ge Zhou Journal: Oncogene Date: 2017-12-22 Impact factor: 9.867