- Email: [email protected]
OP0022 TIE-2 regulates stemness and metastasis of prostate cancer cells
e8
Abstracts / 50 (2014) e1–e74
OP0021 NON-SMALL-CELL LUNG CANCER EXHIBITS POTENT CHARACTERISTICS OF STEM CELLS
MULTI-
N. Zakaria a, N.M. Yusoff a, Z. Zakaria b, M.N. Lim b, P.J.N.M. Baharuddin b, S.K. Fakiruddin b, B. Yahaya a,* . a Cluster for Regenerative Medicine, Advanced Medical and Dental Institute (AMDI), Universiti Sains Malaysia, Malaysia, b Cancer Research Center, Institute for Medical Research (IMR), Kuala Lumpur, Malaysia Background: Lung cancer remains a major cause of cancer-related death because of the high incidence and recurrence despite significant advances in staging and therapies. Recent evidence indicates that tumours contain a small population of cells known as cancer stem cells (CSCs) that are responsible for tumour maintenance and spreading and are resistant to chemotherapy. The genetic composition of CSCs is yet to be fully understood and manipulation of the specific gene that maintains their integrity would be beneficial for future strategies to combat cancer. Methods: In the current study, we isolated putative lung CSCs from a lung adenocarcinoma cell line (A549) and normal stem cells from normal bronchial epithelial cells (PHBEC) on the basis of positive expression of stem cell surface markers (CD166, CD44 and EpCAM) using fluorescence-activated cell sorting. Findings: The putative lung CSC phenotypes of CD166+CD44+ and CD166+EpCAM+ showed multipotent characteristics of stem cells, including the capability to differentiate into osteogenic and chondrogenic, self-renewal, and expression of stem cell transcription factors such as Sox2, Oct3/4, and Klf4. Microarray and bioinformatics data analyses revealed that the putative lung CSCs have molecular signatures for both normal and cancer stem cells with most prominent biological functions associated with angiogenesis, migration, proapoptotic and antiapoptotic osteoblast differentiation, mesenchymal cell differentiation, and mesenchyme development. Additionally, self-renewal pathways such as Wnt and Hedgehog signalling pathways, pathways in cancer, apoptosis, p53, and the ECM-receptor interaction pathway are also significantly associated with the putative CSCs. Interpretation: The study revealed that isolated lung CSCs exhibit the characteristics of multipotent stem cells and their genetic composition might be valuable for future gene and stem cell therapy for lung cancer.
marrow homing and colonisation of haemopoietic stem cells. Notably, this Tie-2-positive population exists exclusively in highly metastatic prostate cancer cell lines. Methods: To study the role of Tie-2 in prostate tumour metastasis, we used fluorescence-activated cell sorting to isolate the Tie-2-positive population from a prostate cancer cell line (PC-3). We then used cDNA microarray analysis to characterise the gene expression profile of these cells. Furthermore, we did a cell adhesion assay to examine the ability of the Tie-2-positive cells in adhering to osteoblasts and endothelial cells. We also undertook quiescent staining and a drug sensitivity assay to examine whether Tie-2-positive cells are more quiescent, and thus become resistant to chemotherapeutic drugs. Finally, we injected both Tie-2-positive and Tie-2-negative PC-3 cells intracardiacly into NOD-SCID mice to establish whether Tie-2 expression promotes prostate tumour metastasis in vivo. Findings: Data from our study revealed that Tie-2-positive cells express higher level of prostate CSC markers compared with the Tie2-negative population. Meanwhile, Tie-2-positive cells are highly adhesive to both osteoblasts and endothelial cells, a characteristic necessary for tumour metastasis. We also found that Tie-2-positive cells are more quiescent and resistant to the chemotherapeutic drug cabazitaxel, providing further support that these cells possess CSC-like characteristics. More importantly, we found that Tie-2-positive cells, but not Tie-2negative cells, developed metastatic tumours in vivo. Interpretation: Our data suggest that Tie-2 plays an important part in the development of drug resistance and prostate tumour metastasis. Thus, Tie-2 might be a novel therapeutic target for treatment of patients with advanced prostate cancer.
http://dx.doi.org/10.1016/j.ejca.2014.03.040
OP0023 CTLA4 SINGLE NUCLEOTIDE POLYMORPHISMS CORRELATE WITH RESPONSE TO BCG IMMUNOTHERAPY R. Mahendran a,b,*, S.M. Tham a, J.N. Rahmat a, Y.K. Lim a, J.H. Sng a,b, L.N. Raman a, Z.M.W. Ma a, W.C. Tsang a, E. Chiong a, Y.H. Chan a, K. Esuvaranathan a . a Department of Surgery, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, b National University Cancer Institute, Singapore
http://dx.doi.org/10.1016/j.ejca.2014.03.039
OP0022 TIE-2 REGULATES STEMNESS PROSTATE CANCER CELLS
AND
METASTASIS
OF
K.D. Tang *, M.-T. Ling. Australia Prostate Cancer Research Centre – Queensland, Institute of Health and Biomedical Innovation, Queensland University of Technology, Princess Alexandra Hospital, Translational Research Institute, Brisbane, QLD, Australia Background: Prostate cancer is the most commonly diagnosed male cancer in western countries. Currently, the major treatment challenge for prostate cancer patients is the development of tumour metastasis. Ample evidence supports the idea that tumour metastasis originates from a rare population of cancer cells known as cancer stem cells (CSCs). Unfortunately, little is known about the identity of these cells, making it difficult to target prostate tumour metastasis. Here we report the identification of a rare population of prostate cancer cells that express the Tie-2 protein, a tyrosine kinase receptor needed for bone
Background: Although BCG is the gold standard for non-muscle invasive bladder cancer, there is a significant non-responder rate. The reasons for this are not known, although single nucleotide polymorphisms (SNPs) in some genes correlate with response to therapy. Bladder cancer patients who were treated with anti-CTLA4 antibodies had increased CD4+ICOShi IFNc expressing T cells over Treg cell numbers in the bladder and peripheral blood. Therefore seven SNPs in the CTLA4 gene (rs733618, rs4553808, rs5742909, rs231775, rs3087243, rs7565213, rs960792) that regulate protein expression and function were chosen for analysis. Methods: Institutional review board approval was obtained for this project. SNPs in 138 bladder cancer patients, who had previously been treated with BCG and for whom long-term follow-up data were available, and 146 healthy controls were analysed. Genomic DNA was extracted and subjected to PCR followed by high resolution melt (HRM) analysis and sequence analyses. Findings: There were no differences in the incidence of SNPs between healthy controls and patients. By Kaplan–Meier analysis, GA at rs4553808 (148 months) and CT at rs5742909 (167 months) correlated with longer time to recurrence. By Cox regression analysis, CT at rs5742909; AA at rs3087243; AA and AG at rs7565213 and TT at
Abstracts / 50 (2014) e1–e74
OP0021 NON-SMALL-CELL LUNG CANCER EXHIBITS POTENT CHARACTERISTICS OF STEM CELLS
MULTI-
N. Zakaria a, N.M. Yusoff a, Z. Zakaria b, M.N. Lim b, P.J.N.M. Baharuddin b, S.K. Fakiruddin b, B. Yahaya a,* . a Cluster for Regenerative Medicine, Advanced Medical and Dental Institute (AMDI), Universiti Sains Malaysia, Malaysia, b Cancer Research Center, Institute for Medical Research (IMR), Kuala Lumpur, Malaysia Background: Lung cancer remains a major cause of cancer-related death because of the high incidence and recurrence despite significant advances in staging and therapies. Recent evidence indicates that tumours contain a small population of cells known as cancer stem cells (CSCs) that are responsible for tumour maintenance and spreading and are resistant to chemotherapy. The genetic composition of CSCs is yet to be fully understood and manipulation of the specific gene that maintains their integrity would be beneficial for future strategies to combat cancer. Methods: In the current study, we isolated putative lung CSCs from a lung adenocarcinoma cell line (A549) and normal stem cells from normal bronchial epithelial cells (PHBEC) on the basis of positive expression of stem cell surface markers (CD166, CD44 and EpCAM) using fluorescence-activated cell sorting. Findings: The putative lung CSC phenotypes of CD166+CD44+ and CD166+EpCAM+ showed multipotent characteristics of stem cells, including the capability to differentiate into osteogenic and chondrogenic, self-renewal, and expression of stem cell transcription factors such as Sox2, Oct3/4, and Klf4. Microarray and bioinformatics data analyses revealed that the putative lung CSCs have molecular signatures for both normal and cancer stem cells with most prominent biological functions associated with angiogenesis, migration, proapoptotic and antiapoptotic osteoblast differentiation, mesenchymal cell differentiation, and mesenchyme development. Additionally, self-renewal pathways such as Wnt and Hedgehog signalling pathways, pathways in cancer, apoptosis, p53, and the ECM-receptor interaction pathway are also significantly associated with the putative CSCs. Interpretation: The study revealed that isolated lung CSCs exhibit the characteristics of multipotent stem cells and their genetic composition might be valuable for future gene and stem cell therapy for lung cancer.
marrow homing and colonisation of haemopoietic stem cells. Notably, this Tie-2-positive population exists exclusively in highly metastatic prostate cancer cell lines. Methods: To study the role of Tie-2 in prostate tumour metastasis, we used fluorescence-activated cell sorting to isolate the Tie-2-positive population from a prostate cancer cell line (PC-3). We then used cDNA microarray analysis to characterise the gene expression profile of these cells. Furthermore, we did a cell adhesion assay to examine the ability of the Tie-2-positive cells in adhering to osteoblasts and endothelial cells. We also undertook quiescent staining and a drug sensitivity assay to examine whether Tie-2-positive cells are more quiescent, and thus become resistant to chemotherapeutic drugs. Finally, we injected both Tie-2-positive and Tie-2-negative PC-3 cells intracardiacly into NOD-SCID mice to establish whether Tie-2 expression promotes prostate tumour metastasis in vivo. Findings: Data from our study revealed that Tie-2-positive cells express higher level of prostate CSC markers compared with the Tie2-negative population. Meanwhile, Tie-2-positive cells are highly adhesive to both osteoblasts and endothelial cells, a characteristic necessary for tumour metastasis. We also found that Tie-2-positive cells are more quiescent and resistant to the chemotherapeutic drug cabazitaxel, providing further support that these cells possess CSC-like characteristics. More importantly, we found that Tie-2-positive cells, but not Tie-2negative cells, developed metastatic tumours in vivo. Interpretation: Our data suggest that Tie-2 plays an important part in the development of drug resistance and prostate tumour metastasis. Thus, Tie-2 might be a novel therapeutic target for treatment of patients with advanced prostate cancer.
http://dx.doi.org/10.1016/j.ejca.2014.03.040
OP0023 CTLA4 SINGLE NUCLEOTIDE POLYMORPHISMS CORRELATE WITH RESPONSE TO BCG IMMUNOTHERAPY R. Mahendran a,b,*, S.M. Tham a, J.N. Rahmat a, Y.K. Lim a, J.H. Sng a,b, L.N. Raman a, Z.M.W. Ma a, W.C. Tsang a, E. Chiong a, Y.H. Chan a, K. Esuvaranathan a . a Department of Surgery, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, b National University Cancer Institute, Singapore
http://dx.doi.org/10.1016/j.ejca.2014.03.039
OP0022 TIE-2 REGULATES STEMNESS PROSTATE CANCER CELLS
AND
METASTASIS
OF
K.D. Tang *, M.-T. Ling. Australia Prostate Cancer Research Centre – Queensland, Institute of Health and Biomedical Innovation, Queensland University of Technology, Princess Alexandra Hospital, Translational Research Institute, Brisbane, QLD, Australia Background: Prostate cancer is the most commonly diagnosed male cancer in western countries. Currently, the major treatment challenge for prostate cancer patients is the development of tumour metastasis. Ample evidence supports the idea that tumour metastasis originates from a rare population of cancer cells known as cancer stem cells (CSCs). Unfortunately, little is known about the identity of these cells, making it difficult to target prostate tumour metastasis. Here we report the identification of a rare population of prostate cancer cells that express the Tie-2 protein, a tyrosine kinase receptor needed for bone
Background: Although BCG is the gold standard for non-muscle invasive bladder cancer, there is a significant non-responder rate. The reasons for this are not known, although single nucleotide polymorphisms (SNPs) in some genes correlate with response to therapy. Bladder cancer patients who were treated with anti-CTLA4 antibodies had increased CD4+ICOShi IFNc expressing T cells over Treg cell numbers in the bladder and peripheral blood. Therefore seven SNPs in the CTLA4 gene (rs733618, rs4553808, rs5742909, rs231775, rs3087243, rs7565213, rs960792) that regulate protein expression and function were chosen for analysis. Methods: Institutional review board approval was obtained for this project. SNPs in 138 bladder cancer patients, who had previously been treated with BCG and for whom long-term follow-up data were available, and 146 healthy controls were analysed. Genomic DNA was extracted and subjected to PCR followed by high resolution melt (HRM) analysis and sequence analyses. Findings: There were no differences in the incidence of SNPs between healthy controls and patients. By Kaplan–Meier analysis, GA at rs4553808 (148 months) and CT at rs5742909 (167 months) correlated with longer time to recurrence. By Cox regression analysis, CT at rs5742909; AA at rs3087243; AA and AG at rs7565213 and TT at