Day :
- Cancer Cell Biology l Cancer Biomarkers l Cancer Vaccines
Session Introduction
Tzu Ching Chang
China Medical University, Taiwan
Title: Mesenchymal stem cells in inflammatory microenvironment promotes cancer cell migration and epithelial-mesenchymal transition through osteopontin
Biography:
Tzu-Ching Chang has completed her PhD from Graduate Institute of Life Science, National Defense Medical Center, Taiwan. She did her Master’s degree from Institute of Physiology, National Taiwan University, College of Medicine, Taiwan. Her areas of research include Molecular Cell Biology, Stem Cell Biology and Cancer Cell Biology.
Abstract:
Backgrounds: Mesenchymal stem cells (MSCs) provide potential regenerative effects on chronic kidney disease (CKD) due to their paracrine signaling with cell tropic and anti-inflammation properties. However, in CKD patients combined with renal cell carcinoma (RCC), the roles of MSCs in inflammatory or tumors’ microenvironment are still controversial. Methods & Results: To evaluate the real characteristics of MSCs in inflammatory microenvironment, we first detected the human MSC secretome by cytokine array after MSCs were incubated with or without inflammatory cytokines (IL-1β or TNF-α). The array data showed that, compared to control MSCs, osteopontin (OPN) were significantly increased after MSCs were incubated with inflammatory cytokines. Furthermore, OPN mRNA level and protein secretion were confirmed by quantitative real-time PCR and ELISA respectively. We further collected MSC condition medium (MSC-CM) after inflammatory cytokine treatments to test the effects of MSC-CM as well as OPN alone on RCC migration. MSC-CM and OPN alone facilitated RCC migration by Boydon chamber assay. The epithelial-mesenchymal transition (EMT) of RCC was significantly promoted by MSC-CM and OPN alone since the reduction of E-cadherin and increase of Snail and vimentin. Using OPN-specific antibody to neutralize OPN in MSC-CM would attenuate the RCC migration and EMT that were stimulated by whole MSC-CM. Moreover, the activities of cancer-associated fibroblasts (CAF) were enhanced by treating CAF with MSC-CM and OPN alone. Conclusion & Significance: In this study, we demonstrated that OPN secreted by MSCs would promote cancer cell migration, EMT progression and CAF activation while MSCs were incubated in an inflammatory microenvironment. We hope that this study will provide another consideration of MSC application in kidney regeneration while CKD patients combined with RCC.
Surangkana Sanmai
Khon Kaen University, Thailand
Title: High expression of Pyruvate Dehydrogenase Kinase in Cholangiocarcinoma patients
Biography:
Surangkana Sanmai is currently pursuing a Master’s degree in Medical Sciences at the Faculty of Associated Medical Sciences, Khon Kaen University, Thailand. She is interested in cancer biomarkers and chemotherapeutic target.
Abstract:
In our previous demonstration, we found the differential mitochondrial proteom between CCA and adjacent non-canerous tissues assessed by mass spectrometry. Cancer cells are well-know for their altered metabolic state and the main cause under this phenomenon is mitochondrial metabolic enzyme permutation. So in this study, the glucose metabolism-involved proteins were selected from data and fold-change analysis was performed. The data showed the overexpression of pyruvate dehydrogenase kinase (PDK) isoform 2 and 3 in CCA compared to its adjacent non-cancerous tissues. Especially, PDK3 showed the highest fold-change at 13. PDK is a Ser/Thr kinase that inactivates mitochondrial pyruvate dehydrogenase and plays a keystone of Warburg effect that lead cancer cells survive in hypoxia condition. PDK isoforms consist of PDK1, PDK2, PDK3 and PDK4. All PDK isoforms notably express on specific tissues and there is no report of PDK pattern in CCA yet. In this work, all PDK isoforms immunohistochemistry was performed in CCA and adjacent non-cancerous tissues to confirm mass spectrometric data. The results showed the pattern of PDK isoform expression where PDK1, PDK2 and PDK3 exhibited high expression on CCA tissues at H-score 240±69, 210±30 and 262±28 respectively (maximum H-score=300), while the weak staining was observed in adjacent tissues. In contrast, PDK4 showed no staining in CCA tissues. Taken together, the specification of PDKs pattern in CCA may provide a good tumor marker for CCA. Futhermore, PDK1 and PDK3 may be considered as potential prognostic markers for CCA patients due to their tumor progression involvement.
Doungdean Tummanatsakun
Khon Kaen University, Thailand
Title: Serum apurinic/apyrimidinic endodeoxyribonuclease 1 in cholangiocarcinoma patients
Biography:
Doungdean Tummanatsakun is currently pursuing a Doctor of Philosophy Program in Medical Technology at the Faculty of Associated Medical Sciences, Khon Kaen University, Thailand. She is interested in cancer biomarkers.
Abstract:
Cholangiocarcinoma (CCA) is a cancer from intra- and extra-hepatic biliary epithelial cells. The incidence of CCA in the northeastern part of Thailand is very high, especially Khon Kaen province. Metastasis of tumor provides important prognostic information and can help guide therapy. This study is to discover CCA prognosis marker to distinguish between non-metastatic and metastatic patients from our database in CCA cell lines secretome. Using bioinformatics include SignalP, SecretomeP, TMHMM, Plasma Proteome Database and EMBOSS for selected candidate proteins from three CCA secretome but not in control immortalized cholangiocyte by GeLC-MS/MS. Secretory protein was selected for further validation studies according to criteria including proteins predicted to have signal peptide or non-classical proteins that have no transmembrane helix, identified in serum and selected proteins with maximum of signal peptide cleavage sites. Apurinic/apyrimidinic endodeoxyribonuclease 1 (APEX1) was selected for further validation analysis. APEX1 has been reported to play a role in metastatic function. Thus, to validate APEX1 protein was differentially expressed in CCA and western blot (WB) for APEX1 was performed for metastatic and non-metastatic group in serum CCA. This revealed that APEX1 intensity ratio was highly expressed in metastatic compared to non-metastatic group (4.73±2.02 for metastatic vs. 1.14±0.81 for non-metastatic group). In conclusion, serum APEX1 might be useful as a prognosis biomarker for treatment in CCA patients.
Daraporn Chua-on
Khon Kaen University, Thailand
Title: High expression of apoptosis-inducing factor, mitochondrion-associated 3 (AIFM3) in human cholangiocarcinoma
Biography:
Daraporn Chua-on is currently pursuing a Doctoral degree in Medical Sciences at the Faculty of Associated Medical Sciences, Khon Kaen University, Thailand. She is interested in cancer biomarkers and chemotherapeutic target.
Abstract:
Diagnostic biomarker of cholangiocarcinoma (CCA) patients is absolutely poor in spite of extensive efforts for the improvment of diagnosis. Mitochondial proteins play key roles in carcinogenesis of various cancer. Therefore, mitochondria are considered as the target organelles for discovering biomarkers of several cancers including CCA. The purpose of this study is to identify potential candidate proteins for diagnosis using mitochondrial proteome analysis for CCA tissues. A shotgun proteomic approch using SDSPAGE coupled with LC-MS/MS was applied to compare the expression of mitochondrial proteins in CCA and the adjacent noncancerous tissues. Using the proteomic analysis for the pooled mitochondrial proteins purified from three each of papillary and nonpapillary types of CCA and their adjacent tissues, 281 proteins were identified as mitochondrial proteins, and 105 of them showed significantly different expression compared with the corresponding counterparts. Among 105 proteins, apoptosis-inducing factor, mitochondrion-associated 3 (AIFM3) was a unique protein commonly over-expressed in both papillary and non-papillary types ot CCA tissues but not in the adjacent non-cancerous tissues. Immunohistochemistry showed expression of AIFM3 staining was significantly higher in CCA than in the corresponding adjacent non-cancerous tissues. Furthermore, high expression of AIFM3 was also significantly expressed in human serum CCA. In conclusion, AIFM3 was aberrantly expressed in both CCA tissues and serum. This finding suggeests that AIFM3 could be a potential biomarker for CCA diagnosis.