{"id":{"repo_id":"ajou","oai_identifier":"oai:repository.ajou.ac.kr:201003/7529"},"canonical_url":"https://search.dev.ndltd.org/etd/ajou/oai:repository.ajou.ac.kr:201003/7529","repository":{"repo_id":"ajou","name":"Ajou University","base_url":"http://repository.ajou.ac.kr/oai/request"},"display":{"title":"Effect of Combination of Aspirin and TRAIL in HeLa Cervical Cancer Cells","abstract":"Purpose: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a member of the tumor necrosis factor (TNF) family of cytokines and induces apoptosis in most tumor cells. Although TRAIL is considered a promising drug for cancer therapies because of its tumor selectivity, many tumors are resistant to TRAIL. Thus TRAIL-resistant cancer cells must be sensitized first to become responsive to TRAIL. In this study, I studied whether pretreatment by aspirin augmented TRAIL-induced apoptotic death in HeLa cells, which derived from cervical cancer and investigated the underlying mechanism. Methods: Cell viability and proliferation were assessed by MTT assay and cytotoxicity was analyzed by LDH assay. Annexin-V /PI staining and sub G1 analysis were used for evaluation of apoptotic cells. We measured mitochondrial membrane potential in HeLa cells undergoing apoptosis by using JC-1. Protein level changes were documented by western blot analysis. Results: Aspirin inhibited proliferation of HeLa cells in the time- and dose-dependent manners. The combined treatment of aspirin with TRAIL strongly enhanced TRAIL-induced apoptotic cell death in HeLa cells. It activated caspase-8, -9 and -3 and caused the caspase-dependent loss of MMP and the release of cytochrome c from mitochondria to cytosol. The apoptotic characteristics enhanced by the combined treatment were inhibited by a pan-caspase inhibitor z-VAD-fmk. Interestingly, TRAIL caused the activation of ERK, whereas the ERK activation was blocked by aspirin. As a result, the activation of ERK decreased and additionally, Mcl-1 also decreased in combination treatment. Conclusion: TRAIL in combination with aspirin significantly increase apoptosis in HeLa cells through caspase-and mitochondrial-dependent pathway. Mechanism of such a promoted apoptotic effect might be associated with that TRAIL-induced ERK activation which triggers survival signal to HeLa cells is blocked by pretreatment of aspirin. Inhibition of ERK activation enhances TRAIL-induced caspase activation, which rapidly cleavages Mcl-1. It could also trigger translocation of cleaved Bid to mitochondria to activate mitochondrial pathway, which also amplifies caspase activation. Eventually, pre-treated aspirin could block the survival signal and enhance caspase-and mitochondrial-dependent apoptotic cell death in HeLa cells treated with TRAIL.","abstract_html":"Purpose: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a member of the tumor necrosis factor (TNF) family of cytokines and induces apoptosis in most tumor cells. Although TRAIL is considered a promising drug for cancer therapies because of its tumor selectivity, many tumors are resistant to TRAIL. Thus TRAIL-resistant cancer cells must be sensitized first to become responsive to TRAIL. In this study, I studied whether pretreatment by aspirin augmented TRAIL-induced apoptotic death in HeLa cells, which derived from cervical cancer and investigated the underlying mechanism. Methods: Cell viability and proliferation were assessed by MTT assay and cytotoxicity was analyzed by LDH assay. Annexin-V /PI staining and sub G1 analysis were used for evaluation of apoptotic cells. We measured mitochondrial membrane potential in HeLa cells undergoing apoptosis by using JC-1. Protein level changes were documented by western blot analysis. Results: Aspirin inhibited proliferation of HeLa cells in the time- and dose-dependent manners. The combined treatment of aspirin with TRAIL strongly enhanced TRAIL-induced apoptotic cell death in HeLa cells. It activated caspase-8, -9 and -3 and caused the caspase-dependent loss of MMP and the release of cytochrome c from mitochondria to cytosol. The apoptotic characteristics enhanced by the combined treatment were inhibited by a pan-caspase inhibitor z-VAD-fmk. Interestingly, TRAIL caused the activation of ERK, whereas the ERK activation was blocked by aspirin. As a result, the activation of ERK decreased and additionally, Mcl-1 also decreased in combination treatment. Conclusion: TRAIL in combination with aspirin significantly increase apoptosis in HeLa cells through caspase-and mitochondrial-dependent pathway. Mechanism of such a promoted apoptotic effect might be associated with that TRAIL-induced ERK activation which triggers survival signal to HeLa cells is blocked by pretreatment of aspirin. Inhibition of ERK activation enhances TRAIL-induced caspase activation, which rapidly cleavages Mcl-1. It could also trigger translocation of cleaved Bid to mitochondria to activate mitochondrial pathway, which also amplifies caspase activation. Eventually, pre-treated aspirin could block the survival signal and enhance caspase-and mitochondrial-dependent apoptotic cell death in HeLa cells treated with TRAIL.","abstract_has_math":false,"creators":["임, 세란"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["장, 영주","대학원 의생명과학과","201024352"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2012,"date_issued":"2012-10-25T04:49:46Z","date_published":"2012-10-25T04:49:46Z","updated_at":"2026-07-24T00:51:39Z","subjects":[],"languages":["en"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000012384","000000012384"],"render_values":[{"text":"http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000012384","href":"http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000012384","code":true},{"text":"000000012384","href":null,"code":true}]}]},"links":{"outbound_url":"http://repository.ajou.ac.kr/handle/201003/7529","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["장, 영주","대학원 의생명과학과","201024352","임, 세란"]},{"key":"dc:creator","label":"Author","values":["임, 세란"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2012-10-25T04:49:46Z","2012"]},{"key":"dc:type","label":"Dc Type","values":["Thesis","Theses"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://repository.ajou.ac.kr/handle/201003/7529","http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000012384","000000012384"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Purpose: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a member of the tumor necrosis factor (TNF) family of cytokines and induces apoptosis in most tumor cells. Although TRAIL is considered a promising drug for cancer therapies because of its tumor selectivity, many tumors are resistant to TRAIL. Thus TRAIL-resistant cancer cells must be sensitized first to become responsive to TRAIL. In this study, I studied whether pretreatment by aspirin augmented TRAIL-induced apoptotic death in HeLa cells, which derived from cervical cancer and investigated the underlying mechanism. Methods: Cell viability and proliferation were assessed by MTT assay and cytotoxicity was analyzed by LDH assay. Annexin-V /PI staining and sub G1 analysis were used for evaluation of apoptotic cells. We measured mitochondrial membrane potential in HeLa cells undergoing apoptosis by using JC-1. Protein level changes were documented by western blot analysis. Results: Aspirin inhibited proliferation of HeLa cells in the time- and dose-dependent manners. The combined treatment of aspirin with TRAIL strongly enhanced TRAIL-induced apoptotic cell death in HeLa cells. It activated caspase-8, -9 and -3 and caused the caspase-dependent loss of MMP and the release of cytochrome c from mitochondria to cytosol. The apoptotic characteristics enhanced by the combined treatment were inhibited by a pan-caspase inhibitor z-VAD-fmk. Interestingly, TRAIL caused the activation of ERK, whereas the ERK activation was blocked by aspirin. As a result, the activation of ERK decreased and additionally, Mcl-1 also decreased in combination treatment. Conclusion: TRAIL in combination with aspirin significantly increase apoptosis in HeLa cells through caspase-and mitochondrial-dependent pathway. Mechanism of such a promoted apoptotic effect might be associated with that TRAIL-induced ERK activation which triggers survival signal to HeLa cells is blocked by pretreatment of aspirin. Inhibition of ERK activation enhances TRAIL-induced caspase activation, which rapidly cleavages Mcl-1. It could also trigger translocation of cleaved Bid to mitochondria to activate mitochondrial pathway, which also amplifies caspase activation. Eventually, pre-treated aspirin could block the survival signal and enhance caspase-and mitochondrial-dependent apoptotic cell death in HeLa cells treated with TRAIL.","ABSTRACT ⅰ TABLE OF CONTENTS ⅲ LIST OF FIGURES ⅴ Ⅰ. INTRODUCTION 1 Ⅱ. MATERIALS AND METHODS 3 A. Reagents 3 B. Antibodies 3 C. Cell culture 3 D. Cell viability assay 3 E. Cytotoxicity assay 4 F. Sub G1 analyses 4 G. Annexin-V/ PI staining 4 H. Immunoblotting 5 I. Measurement of mitochondrial membrane potential 5 J. Analysis of cytochrome c release 6 Ⅲ. RESULTS 7 1. Aspirin inhibits the proliferation of HeLa cells and augments TRAIL-mediated cell death 7 2. Aspirin triggers TRAIL-induced apoptotic cell death via enhanced caspase activity 10 3. Aspirin results in TRAIL-induced caspase-dependent mitochondrial membrane potential change and cytochrome c release 16 4. Aspirin makes TRAIL-resistant HeLa cells TRAIL-sensitive through inhibition of ERK1/2 activation 19 Ⅳ. DISCUSSION 26 Ⅴ. REFERENCES 29 Ⅵ. 국문요약 36","Master"]},{"key":"dc:title","label":"Title","values":["Effect of Combination of Aspirin and TRAIL in HeLa Cervical Cancer Cells","Aspirin과 TRAIL 병합처리시 자궁경부암인 HeLa cell에 미치는 영향"]}]}],"canonical_facts":{"dc:contributor":["장, 영주","대학원 의생명과학과","201024352","임, 세란"],"dc:creator":["임, 세란"],"dc:date":["2012-10-25T04:49:46Z","2012"],"dc:description":["Purpose: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a member of the tumor necrosis factor (TNF) family of cytokines and induces apoptosis in most tumor cells. Although TRAIL is considered a promising drug for cancer therapies because of its tumor selectivity, many tumors are resistant to TRAIL. Thus TRAIL-resistant cancer cells must be sensitized first to become responsive to TRAIL. In this study, I studied whether pretreatment by aspirin augmented TRAIL-induced apoptotic death in HeLa cells, which derived from cervical cancer and investigated the underlying mechanism. Methods: Cell viability and proliferation were assessed by MTT assay and cytotoxicity was analyzed by LDH assay. Annexin-V /PI staining and sub G1 analysis were used for evaluation of apoptotic cells. We measured mitochondrial membrane potential in HeLa cells undergoing apoptosis by using JC-1. Protein level changes were documented by western blot analysis. Results: Aspirin inhibited proliferation of HeLa cells in the time- and dose-dependent manners. The combined treatment of aspirin with TRAIL strongly enhanced TRAIL-induced apoptotic cell death in HeLa cells. It activated caspase-8, -9 and -3 and caused the caspase-dependent loss of MMP and the release of cytochrome c from mitochondria to cytosol. The apoptotic characteristics enhanced by the combined treatment were inhibited by a pan-caspase inhibitor z-VAD-fmk. Interestingly, TRAIL caused the activation of ERK, whereas the ERK activation was blocked by aspirin. As a result, the activation of ERK decreased and additionally, Mcl-1 also decreased in combination treatment. Conclusion: TRAIL in combination with aspirin significantly increase apoptosis in HeLa cells through caspase-and mitochondrial-dependent pathway. Mechanism of such a promoted apoptotic effect might be associated with that TRAIL-induced ERK activation which triggers survival signal to HeLa cells is blocked by pretreatment of aspirin. Inhibition of ERK activation enhances TRAIL-induced caspase activation, which rapidly cleavages Mcl-1. It could also trigger translocation of cleaved Bid to mitochondria to activate mitochondrial pathway, which also amplifies caspase activation. Eventually, pre-treated aspirin could block the survival signal and enhance caspase-and mitochondrial-dependent apoptotic cell death in HeLa cells treated with TRAIL.","ABSTRACT ⅰ TABLE OF CONTENTS ⅲ LIST OF FIGURES ⅴ Ⅰ. INTRODUCTION 1 Ⅱ. MATERIALS AND METHODS 3 A. Reagents 3 B. Antibodies 3 C. Cell culture 3 D. Cell viability assay 3 E. Cytotoxicity assay 4 F. Sub G1 analyses 4 G. Annexin-V/ PI staining 4 H. Immunoblotting 5 I. Measurement of mitochondrial membrane potential 5 J. Analysis of cytochrome c release 6 Ⅲ. RESULTS 7 1. Aspirin inhibits the proliferation of HeLa cells and augments TRAIL-mediated cell death 7 2. Aspirin triggers TRAIL-induced apoptotic cell death via enhanced caspase activity 10 3. Aspirin results in TRAIL-induced caspase-dependent mitochondrial membrane potential change and cytochrome c release 16 4. Aspirin makes TRAIL-resistant HeLa cells TRAIL-sensitive through inhibition of ERK1/2 activation 19 Ⅳ. DISCUSSION 26 Ⅴ. REFERENCES 29 Ⅵ. 국문요약 36","Master"],"dc:identifier":["http://repository.ajou.ac.kr/handle/201003/7529","http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000012384","000000012384"],"dc:language":["en"],"dc:title":["Effect of Combination of Aspirin and TRAIL in HeLa Cervical Cancer Cells","Aspirin과 TRAIL 병합처리시 자궁경부암인 HeLa cell에 미치는 영향"],"dc:type":["Thesis","Theses"]},"updated_at":"2026-07-24T00:51:39Z"}