{"id":{"repo_id":"adelaide","oai_identifier":"oai:digital.library.adelaide.edu.au:2440/151383"},"canonical_url":"https://search.dev.ndltd.org/etd/adelaide/oai:digital.library.adelaide.edu.au:2440/151383","repository":{"repo_id":"adelaide","name":"University of Adelaide","base_url":"https://digital.library.adelaide.edu.au/server/oai/request"},"display":{"title":"Developing Novel Conservative Treatments For 2 Endometrial Cancer By Targeting Aquaporin-1","abstract":"Endometrial cancer (EC) is sixth most common cancer among females worldwide and 186 the most common invasive gynaecological malignancy. Pathogenesis of endometrial cancer is related to prolonged elevated levels of oestrogen due to obesity, nulliparity, and polycystic ovaries leading to uncontrolled proliferation of the endometrium, resulting in endometrial hyperplasia and cancer. Standard treatment is limited to surgical removal of uterus and bilateral ovaries in low grade cancers. Progestin therapy, a fertility-preserving treatment, is recommended for young women diagnosed with EC who wish to preserve their fertility. Yet, according to ESMO (European Society for Medical Oncology) guidelines, these women are required to undergo removal of uterus once the family is completed. Depriving women of childbearing ability can lead to significant psychological and sexual distress mainly due to loss of fertility and onset of menopausal symptoms. These treatment challenges highlight the urgent need for effective, non-surgical, and fertility-preserving treatments. A growing area of research aims to improve cancer invasiveness by pharmacologically targeting aquaporin 1, a transmembrane protein serving as dual water and ion channels and demonstrating high expression across various solid cancer types. This thesis uses two approaches to identify novel conservative treatment of EC. In the first approach, we tested a library of pharmacological modulators of aquaporins (AQPs) on invasiveness of endometrial cancer cells. In the second approach, we tested Traditional Chinese Medicines (TCMs) that haven’t been previously evaluated for potential to block AQP channels. Additionally, we tested the efficacy of TCMs in reducing the invasiveness of endometrial cancer cells and identified a novel AQP1 ion channel inhibitor. We used Ishikawa and MFE-280 cell lines corresponding to low- and high-grade endometrial cancer cells and primary endometrial cancer cells derived from low- and high- grade cancer tissues. Chapter 3 of this thesis demonstrates that the expression of oestrogen receptors was confirmed in both cell lines and later, the cell lines were treated with a physiological dose of oestradiol (100pM or 1nM) to assess the expression of AQP 0- AQP12b using quantitative polymerase chain reaction (qPCR). High expression of AQP1 transcript was found in both cell lines. Immunostaining of endometrial cancer tissues from patients demonstrated a high expression of AQP1 protein. Western blot analysis further indicated AQP1 protein in both cell lines, which was upregulated by oestradiol at 100pM or 1nM doses. Next, we tested a library of pharmacological inhibitors of AQPs including water channel inhibitors (Acetazolamide 1 μM and 10 μM, Ginsenoside Rg3 100 μM, KeenMind 33 μM), ion channel inhibitors (Bumetanide derivatives AqB011 60 μM-100 μM, 5-HMF, 1mM, 5-PMFC 0.5 mM), and non-specific AQP4 modulators (IMD-0354 0.2 μM, TGN-020 3 μM). A wide range of dose of all drugs were tested and optimal doses causing 50% inhibition of invasion were identified. We found that AqB011 (80 μM) and 5-PMFC (0.5 mM) caused significant reduction of invasion in low- and high-grade endometrial cancer cells in transwell invasion assays. AqB011 and 5-PMFC were further tested in primary endometrial cancer cells derived from patient tissues, AqB011 reduced invasion by 15-50%, while 5-PMFC reduced invasion by 50% in both cell lines. Tissue levels of AQP1 in cancer samples correlated positively with degree of inhibition of invasion by AqB011. The doses of AQB011 and 5-PMFC effective in reducing the invasiveness of cancer cells were tested for cytotoxicity using cell viability assays. The anti- invasive doses did not cause cell death indicating that reduction in invasion of cancer cells is not due to cell death. We also tested library of TCMs; (Resveratrol 40 μM and Curcumin 20 μM previously proven effective in reducing the invasiveness of endometrial cancer cell lines and Guizhi Fuling, under clinical trial for the management of endometriosis) that have not been tested for AQP blocking action on invasiveness of endometrial cancer cells. In chapter 4, we demonstrated that Guizhi Fuling (referred in the thesis as Fuling), significantly reduced the invasiveness of low- and high-grade endometrial cancer cell lines at doses that are tested in clinical trials for endometriosis. The effective dose of Fuling (80 μg/ml) was not toxic to cancer cells as assessed by cell viability assay. Fuling was demonstrated to block the invasive effects of oestradiol. In chapter 5, we confirmed the anti-invasive action of Fuling at non-toxic doses using an in vivo CAM model and on primary cancer cells derived from patient tissues. By electrophysiology we demonstrated that Fuling inhibited ion channel activity, but not the water channel activity of AQP1 expressed in Xenopus oocytes. In summary, the findings of this thesis provide a strong foundation for using AqB011, 5-PMFC, and Fuling as promising candidates for novel conservative therapy in endometrial cancer. Importantly, the thesis introduces a new compound to the library of pharmacological inhibitors targeting AQP1.","abstract_html":"Endometrial cancer (EC) is sixth most common cancer among females worldwide and 186 the most common invasive gynaecological malignancy. Pathogenesis of endometrial cancer is related to prolonged elevated levels of oestrogen due to obesity, nulliparity, and polycystic ovaries leading to uncontrolled proliferation of the endometrium, resulting in endometrial hyperplasia and cancer. Standard treatment is limited to surgical removal of uterus and bilateral ovaries in low grade cancers. Progestin therapy, a fertility-preserving treatment, is recommended for young women diagnosed with EC who wish to preserve their fertility. Yet, according to ESMO (European Society for Medical Oncology) guidelines, these women are required to undergo removal of uterus once the family is completed. Depriving women of childbearing ability can lead to significant psychological and sexual distress mainly due to loss of fertility and onset of menopausal symptoms. These treatment challenges highlight the urgent need for effective, non-surgical, and fertility-preserving treatments. A growing area of research aims to improve cancer invasiveness by pharmacologically targeting aquaporin 1, a transmembrane protein serving as dual water and ion channels and demonstrating high expression across various solid cancer types. This thesis uses two approaches to identify novel conservative treatment of EC. In the first approach, we tested a library of pharmacological modulators of aquaporins (AQPs) on invasiveness of endometrial cancer cells. In the second approach, we tested Traditional Chinese Medicines (TCMs) that haven’t been previously evaluated for potential to block AQP channels. Additionally, we tested the efficacy of TCMs in reducing the invasiveness of endometrial cancer cells and identified a novel AQP1 ion channel inhibitor. We used Ishikawa and MFE-280 cell lines corresponding to low- and high-grade endometrial cancer cells and primary endometrial cancer cells derived from low- and high- grade cancer tissues. Chapter 3 of this thesis demonstrates that the expression of oestrogen receptors was confirmed in both cell lines and later, the cell lines were treated with a physiological dose of oestradiol (100pM or 1nM) to assess the expression of AQP 0- AQP12b using quantitative polymerase chain reaction (qPCR). High expression of AQP1 transcript was found in both cell lines. Immunostaining of endometrial cancer tissues from patients demonstrated a high expression of AQP1 protein. Western blot analysis further indicated AQP1 protein in both cell lines, which was upregulated by oestradiol at 100pM or 1nM doses. Next, we tested a library of pharmacological inhibitors of AQPs including water channel inhibitors (Acetazolamide 1 μM and 10 μM, Ginsenoside Rg3 100 μM, KeenMind 33 μM), ion channel inhibitors (Bumetanide derivatives AqB011 60 μM-100 μM, 5-HMF, 1mM, 5-PMFC 0.5 mM), and non-specific AQP4 modulators (IMD-0354 0.2 μM, TGN-020 3 μM). A wide range of dose of all drugs were tested and optimal doses causing 50% inhibition of invasion were identified. We found that AqB011 (80 μM) and 5-PMFC (0.5 mM) caused significant reduction of invasion in low- and high-grade endometrial cancer cells in transwell invasion assays. AqB011 and 5-PMFC were further tested in primary endometrial cancer cells derived from patient tissues, AqB011 reduced invasion by 15-50%, while 5-PMFC reduced invasion by 50% in both cell lines. Tissue levels of AQP1 in cancer samples correlated positively with degree of inhibition of invasion by AqB011. The doses of AQB011 and 5-PMFC effective in reducing the invasiveness of cancer cells were tested for cytotoxicity using cell viability assays. The anti- invasive doses did not cause cell death indicating that reduction in invasion of cancer cells is not due to cell death. We also tested library of TCMs; (Resveratrol 40 μM and Curcumin 20 μM previously proven effective in reducing the invasiveness of endometrial cancer cell lines and Guizhi Fuling, under clinical trial for the management of endometriosis) that have not been tested for AQP blocking action on invasiveness of endometrial cancer cells. In chapter 4, we demonstrated that Guizhi Fuling (referred in the thesis as Fuling), significantly reduced the invasiveness of low- and high-grade endometrial cancer cell lines at doses that are tested in clinical trials for endometriosis. The effective dose of Fuling (80 μg/ml) was not toxic to cancer cells as assessed by cell viability assay. Fuling was demonstrated to block the invasive effects of oestradiol. In chapter 5, we confirmed the anti-invasive action of Fuling at non-toxic doses using an in vivo CAM model and on primary cancer cells derived from patient tissues. By electrophysiology we demonstrated that Fuling inhibited ion channel activity, but not the water channel activity of AQP1 expressed in Xenopus oocytes. In summary, the findings of this thesis provide a strong foundation for using AqB011, 5-PMFC, and Fuling as promising candidates for novel conservative therapy in endometrial cancer. Importantly, the thesis introduces a new compound to the library of pharmacological inhibitors targeting AQP1.","abstract_has_math":false,"creators":["Khan, Sidra Nawaz"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Yool, Andrea","Ricciardelli, Carmela"],"committee_chairs":[],"committee_members":[],"year":2024,"date_issued":"2024","date_published":"2024","updated_at":"2026-08-21T16:41:36Z","subjects":["Ednometrial cancer","Gynaecological cancer","Reproductive health","Women's health","Drug discovery","Aquaporin-1","AQP-1","Guizhi Fuling","Fuling","CAM","AqB011","5-PMFC","Ion channel inhibitors","TCMs"],"languages":["en"],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://hdl.handle.net/2440/151383","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"source_record":{"url":"https://digital.library.adelaide.edu.au/server/oai/request?verb=GetRecord&metadataPrefix=dim&identifier=oai%3Adigital.library.adelaide.edu.au%3A2440%2F151383","prefix":"dim"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Yool, Andrea","Ricciardelli, Carmela"]},{"key":"dc:creator","label":"Author","values":["Khan, Sidra Nawaz"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.issued","label":"Date","values":["2024"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Ednometrial cancer","Gynaecological cancer","Reproductive health","Women's health","Drug discovery","Aquaporin-1","AQP-1","Guizhi Fuling","Fuling","CAM","AqB011","5-PMFC","Ion channel inhibitors","TCMs"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://hdl.handle.net/2440/151383"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Endometrial cancer (EC) is sixth most common cancer among females worldwide and 186 the most common invasive gynaecological malignancy. Pathogenesis of endometrial cancer is related to prolonged elevated levels of oestrogen due to obesity, nulliparity, and polycystic ovaries leading to uncontrolled proliferation of the endometrium, resulting in endometrial hyperplasia and cancer. Standard treatment is limited to surgical removal of uterus and bilateral ovaries in low grade cancers. Progestin therapy, a fertility-preserving treatment, is recommended for young women diagnosed with EC who wish to preserve their fertility. Yet, according to ESMO (European Society for Medical Oncology) guidelines, these women are required to undergo removal of uterus once the family is completed. Depriving women of childbearing ability can lead to significant psychological and sexual distress mainly due to loss of fertility and onset of menopausal symptoms. These treatment challenges highlight the urgent need for effective, non-surgical, and fertility-preserving treatments. A growing area of research aims to improve cancer invasiveness by pharmacologically targeting aquaporin 1, a transmembrane protein serving as dual water and ion channels and demonstrating high expression across various solid cancer types. This thesis uses two approaches to identify novel conservative treatment of EC. In the first approach, we tested a library of pharmacological modulators of aquaporins (AQPs) on invasiveness of endometrial cancer cells. In the second approach, we tested Traditional Chinese Medicines (TCMs) that haven’t been previously evaluated for potential to block AQP channels. Additionally, we tested the efficacy of TCMs in reducing the invasiveness of endometrial cancer cells and identified a novel AQP1 ion channel inhibitor. We used Ishikawa and MFE-280 cell lines corresponding to low- and high-grade endometrial cancer cells and primary endometrial cancer cells derived from low- and high- grade cancer tissues. Chapter 3 of this thesis demonstrates that the expression of oestrogen receptors was confirmed in both cell lines and later, the cell lines were treated with a physiological dose of oestradiol (100pM or 1nM) to assess the expression of AQP 0- AQP12b using quantitative polymerase chain reaction (qPCR). High expression of AQP1 transcript was found in both cell lines. Immunostaining of endometrial cancer tissues from patients demonstrated a high expression of AQP1 protein. Western blot analysis further indicated AQP1 protein in both cell lines, which was upregulated by oestradiol at 100pM or 1nM doses. Next, we tested a library of pharmacological inhibitors of AQPs including water channel inhibitors (Acetazolamide 1 μM and 10 μM, Ginsenoside Rg3 100 μM, KeenMind 33 μM), ion channel inhibitors (Bumetanide derivatives AqB011 60 μM-100 μM, 5-HMF, 1mM, 5-PMFC 0.5 mM), and non-specific AQP4 modulators (IMD-0354 0.2 μM, TGN-020 3 μM). A wide range of dose of all drugs were tested and optimal doses causing 50% inhibition of invasion were identified. We found that AqB011 (80 μM) and 5-PMFC (0.5 mM) caused significant reduction of invasion in low- and high-grade endometrial cancer cells in transwell invasion assays. AqB011 and 5-PMFC were further tested in primary endometrial cancer cells derived from patient tissues, AqB011 reduced invasion by 15-50%, while 5-PMFC reduced invasion by 50% in both cell lines. Tissue levels of AQP1 in cancer samples correlated positively with degree of inhibition of invasion by AqB011. The doses of AQB011 and 5-PMFC effective in reducing the invasiveness of cancer cells were tested for cytotoxicity using cell viability assays. The anti- invasive doses did not cause cell death indicating that reduction in invasion of cancer cells is not due to cell death. We also tested library of TCMs; (Resveratrol 40 μM and Curcumin 20 μM previously proven effective in reducing the invasiveness of endometrial cancer cell lines and Guizhi Fuling, under clinical trial for the management of endometriosis) that have not been tested for AQP blocking action on invasiveness of endometrial cancer cells. In chapter 4, we demonstrated that Guizhi Fuling (referred in the thesis as Fuling), significantly reduced the invasiveness of low- and high-grade endometrial cancer cell lines at doses that are tested in clinical trials for endometriosis. The effective dose of Fuling (80 μg/ml) was not toxic to cancer cells as assessed by cell viability assay. Fuling was demonstrated to block the invasive effects of oestradiol. In chapter 5, we confirmed the anti-invasive action of Fuling at non-toxic doses using an in vivo CAM model and on primary cancer cells derived from patient tissues. By electrophysiology we demonstrated that Fuling inhibited ion channel activity, but not the water channel activity of AQP1 expressed in Xenopus oocytes. In summary, the findings of this thesis provide a strong foundation for using AqB011, 5-PMFC, and Fuling as promising candidates for novel conservative therapy in endometrial cancer. Importantly, the thesis introduces a new compound to the library of pharmacological inhibitors targeting AQP1."]},{"key":"dc:title","label":"Title","values":["Developing Novel Conservative Treatments For 2 Endometrial Cancer By Targeting Aquaporin-1"]}]}],"canonical_facts":{"dc:contributor.advisor":["Yool, Andrea","Ricciardelli, Carmela"],"dc:creator":["Khan, Sidra Nawaz"],"dc:date.issued":["2024"],"dc:description.abstract":["Endometrial cancer (EC) is sixth most common cancer among females worldwide and 186 the most common invasive gynaecological malignancy. Pathogenesis of endometrial cancer is related to prolonged elevated levels of oestrogen due to obesity, nulliparity, and polycystic ovaries leading to uncontrolled proliferation of the endometrium, resulting in endometrial hyperplasia and cancer. Standard treatment is limited to surgical removal of uterus and bilateral ovaries in low grade cancers. Progestin therapy, a fertility-preserving treatment, is recommended for young women diagnosed with EC who wish to preserve their fertility. Yet, according to ESMO (European Society for Medical Oncology) guidelines, these women are required to undergo removal of uterus once the family is completed. Depriving women of childbearing ability can lead to significant psychological and sexual distress mainly due to loss of fertility and onset of menopausal symptoms. These treatment challenges highlight the urgent need for effective, non-surgical, and fertility-preserving treatments. A growing area of research aims to improve cancer invasiveness by pharmacologically targeting aquaporin 1, a transmembrane protein serving as dual water and ion channels and demonstrating high expression across various solid cancer types. This thesis uses two approaches to identify novel conservative treatment of EC. In the first approach, we tested a library of pharmacological modulators of aquaporins (AQPs) on invasiveness of endometrial cancer cells. In the second approach, we tested Traditional Chinese Medicines (TCMs) that haven’t been previously evaluated for potential to block AQP channels. Additionally, we tested the efficacy of TCMs in reducing the invasiveness of endometrial cancer cells and identified a novel AQP1 ion channel inhibitor. We used Ishikawa and MFE-280 cell lines corresponding to low- and high-grade endometrial cancer cells and primary endometrial cancer cells derived from low- and high- grade cancer tissues. Chapter 3 of this thesis demonstrates that the expression of oestrogen receptors was confirmed in both cell lines and later, the cell lines were treated with a physiological dose of oestradiol (100pM or 1nM) to assess the expression of AQP 0- AQP12b using quantitative polymerase chain reaction (qPCR). High expression of AQP1 transcript was found in both cell lines. Immunostaining of endometrial cancer tissues from patients demonstrated a high expression of AQP1 protein. Western blot analysis further indicated AQP1 protein in both cell lines, which was upregulated by oestradiol at 100pM or 1nM doses. Next, we tested a library of pharmacological inhibitors of AQPs including water channel inhibitors (Acetazolamide 1 μM and 10 μM, Ginsenoside Rg3 100 μM, KeenMind 33 μM), ion channel inhibitors (Bumetanide derivatives AqB011 60 μM-100 μM, 5-HMF, 1mM, 5-PMFC 0.5 mM), and non-specific AQP4 modulators (IMD-0354 0.2 μM, TGN-020 3 μM). A wide range of dose of all drugs were tested and optimal doses causing 50% inhibition of invasion were identified. We found that AqB011 (80 μM) and 5-PMFC (0.5 mM) caused significant reduction of invasion in low- and high-grade endometrial cancer cells in transwell invasion assays. AqB011 and 5-PMFC were further tested in primary endometrial cancer cells derived from patient tissues, AqB011 reduced invasion by 15-50%, while 5-PMFC reduced invasion by 50% in both cell lines. Tissue levels of AQP1 in cancer samples correlated positively with degree of inhibition of invasion by AqB011. The doses of AQB011 and 5-PMFC effective in reducing the invasiveness of cancer cells were tested for cytotoxicity using cell viability assays. The anti- invasive doses did not cause cell death indicating that reduction in invasion of cancer cells is not due to cell death. We also tested library of TCMs; (Resveratrol 40 μM and Curcumin 20 μM previously proven effective in reducing the invasiveness of endometrial cancer cell lines and Guizhi Fuling, under clinical trial for the management of endometriosis) that have not been tested for AQP blocking action on invasiveness of endometrial cancer cells. In chapter 4, we demonstrated that Guizhi Fuling (referred in the thesis as Fuling), significantly reduced the invasiveness of low- and high-grade endometrial cancer cell lines at doses that are tested in clinical trials for endometriosis. The effective dose of Fuling (80 μg/ml) was not toxic to cancer cells as assessed by cell viability assay. Fuling was demonstrated to block the invasive effects of oestradiol. In chapter 5, we confirmed the anti-invasive action of Fuling at non-toxic doses using an in vivo CAM model and on primary cancer cells derived from patient tissues. By electrophysiology we demonstrated that Fuling inhibited ion channel activity, but not the water channel activity of AQP1 expressed in Xenopus oocytes. In summary, the findings of this thesis provide a strong foundation for using AqB011, 5-PMFC, and Fuling as promising candidates for novel conservative therapy in endometrial cancer. Importantly, the thesis introduces a new compound to the library of pharmacological inhibitors targeting AQP1."],"dc:identifier.uri":["https://hdl.handle.net/2440/151383"],"dc:language.iso":["en"],"dc:subject":["Ednometrial cancer","Gynaecological cancer","Reproductive health","Women's health","Drug discovery","Aquaporin-1","AQP-1","Guizhi Fuling","Fuling","CAM","AqB011","5-PMFC","Ion channel inhibitors","TCMs"],"dc:title":["Developing Novel Conservative Treatments For 2 Endometrial Cancer By Targeting Aquaporin-1"],"dc:type":["Thesis"]},"updated_at":"2026-08-21T16:41:36Z"}