2018_Cytotoxic Evaluation of Orthosiphon Stamineus Benth Extracts and Their Molecular Mechanism of Apoptosis on Uterine Leiomyosarcoma (SK-UT-1) Cells
| Format: | General Document |
|---|
| _version_ | 1860798020563501056 |
|---|---|
| building | INTELEK Repository |
| collection | Online Access |
| collectionurl | https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection3 |
| copyright | Copyright©PWB2025 |
| country | Malaysia |
| date | 2018-08-12 |
| format | General Document |
| id | 15501 |
| institution | UniSZA |
| internalnotes | Sila masukkan subject wajib Dissertations, Academic untuk semua tesis.. Terima kasih |
| originalfilename | CYTOTOXIC EVALUATION OF ORTHOSIPHON STAMINEUS BENTH EXTRACTS AND THEIR MOLECULAR MECHANISM OF APOPTOSIS ON UTERINE LEIOMYOSARCOMA (SK-UT-1) CELLS (MASTER_2018).pdf |
| person | Norzilawati Pauzi |
| recordtype | oai_dc |
| resourceurl | https://intelek.unisza.edu.my/intelek/pages/view.php?ref=15501 |
| sourcemedia | Server storage Scanned document |
| spelling | 15501 https://intelek.unisza.edu.my/intelek/pages/view.php?ref=15501 https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection3 General Document Malaysia Library Staff (Top Management) Library Staff (Management) Library Staff (Support) Terengganu Faculty of Bio-resources & Food Industry English application/pdf 1.5 Server storage Scanned document Universiti Sultan Zainal Abidin UniSZA Private Access UNIVERSITI SULTAN ZAINAL ABIDIN SAMBox 2.3.4; modified using iTextSharp™ 5.5.10 ©2000-2016 iText Group NV (AGPL-version) Norzilawati Pauzi 202 CYTOTOXIC EVALUATION OF ORTHOSIPHON STAMINEUS BENTH EXTRACTS AND THEIR MOLECULAR MECHANISM OF APOPTOSIS ON UTERINE LEIOMYOSARCOMA (SK-UT-1) CELLS (MASTER_2018).pdf Copyright©PWB2025 2018-08-12 2018_Cytotoxic Evaluation of Orthosiphon Stamineus Benth Extracts and Their Molecular Mechanism of Apoptosis on Uterine Leiomyosarcoma (SK-UT-1) Cells Uterine leiomyosarcoma is a rare malignant tumor that arises from smooth muscle of the uterus. Uterine leiomyosarcoma shared similarities with uterine fibroid which regulated by several growth factors which caused an excessive menstrual bleeding and miscarriage. It contributes to the most hysterectomy cases, a surgery which results in complete removal of the uterus. Orthosiphon stamineus Benth (Misai Kucing) is a herbal plant originated from South East Asia including Malaysia. Several studies have been conducted and many findings have been reported about the efficacy of O. stamineus in biological activities including antioxidant, antitumor and antiangiogenesis. However, their activity against uterine leiomyosarcoma (SK-UT-1) cells has not yet explored. The present study was conducted to investigate the effect of water (OSW) and ethanol (50%) (OSEW) O. stamineus extracts against uterine leiomyosarcoma (SK-UT-1) cells by determining the cytotoxicity and cell cycle effect, assessing the mode of cell death and evaluating the effect of the extracts on the growth factor via gene expression. The cytotoxicity effect was conducted using MTT assay whereas BD CycleTestTM Plus DNA Kit assay was carried out to determine the effect of extracts on cell cycle of SK-UT-1 cells. Their effect on cell cycle-related (p53 and Cyclin B1) and inflammation-related (PARP-1 and Iκβα) proteins expression were assessed using Western blot. The presence of apoptotic cell morphologies and their population was determined using double staining assays (AOPI and Annexin V FITC/PI). Real-time reverse transcription polymerase chain reaction (qRT-PCR) was implemented to assess the effect of O. stamineus extracts on apoptotic-related (BAX and BCL-2) and growth factor-related (VEGF-A, TGF-β1 and PCNA) genes expression. The results showed that OSW and OSEW have moderate and no cytotoxic effect against SK-UT-1 cells with IC50 values 18 μg/mL and 34 μg/mL, respectively. Both extracts showed no cytotoxic effects towards normal fibroblast (NIH/3T3) cells with IC50 exceed 100 μg/mL. The extracts were also found to induce cell cycle arrest at G0/G1 by OSW and G0/G1 and G2/M by OSEW by regulating the Cyclin B1 protein expression in a p53-independent manner. AO/PI staining assay showed the presence of apoptotic cell morphologies and their population in Annexin V-FITC/PI assay were increased in dose-dependent manner. qRT-PCR analysis using relative quantitative method revealed the up regulation of BAX by OSEW alone and downregulation of BCL-2 by both extracts. Western blotting revealed the presence of PARP-1 cleavage and the increase of Iκβα protein expression by both extracts. Apart from that, VEGF A, TGF-β1 and PCNA genes expression were found to be downregulated by both extracts. From this study, it can be concluded that the O. stamineus extracts possess moderate cytotoxic, anti-proliferative and anti-inflammatory effect through regulation at biochemical level expression. It also induced apoptosis in uterine leiomyosarcoma cells (SK-UT-1) in a dose-dependent manner through intrinsic pathway. Orthosiphon stamineus Dissertations, Academic Sila masukkan subject wajib Dissertations, Academic untuk semua tesis.. Terima kasih Apoptosis Induction Orthosiphon Stamineus Cytotoxic Evaluation Thesis |
| spellingShingle | 2018_Cytotoxic Evaluation of Orthosiphon Stamineus Benth Extracts and Their Molecular Mechanism of Apoptosis on Uterine Leiomyosarcoma (SK-UT-1) Cells |
| state | Terengganu |
| subject | Orthosiphon stamineus Dissertations, Academic |
| summary | Uterine leiomyosarcoma is a rare malignant tumor that arises from smooth muscle of the uterus. Uterine leiomyosarcoma shared similarities with uterine fibroid which regulated by several growth factors which caused an excessive menstrual bleeding and miscarriage. It contributes to the most hysterectomy cases, a surgery which results in complete removal of the uterus. Orthosiphon stamineus Benth (Misai Kucing) is a herbal plant originated from South East Asia including Malaysia. Several studies have been conducted and many findings have been reported about the efficacy of O. stamineus in biological activities including antioxidant, antitumor and antiangiogenesis. However, their activity against uterine leiomyosarcoma (SK-UT-1) cells has not yet explored. The present study was conducted to investigate the effect of water (OSW) and ethanol (50%) (OSEW) O. stamineus extracts against uterine leiomyosarcoma (SK-UT-1) cells by determining the cytotoxicity and cell cycle effect, assessing the mode of cell death and evaluating the effect of the extracts on the growth factor via gene expression. The cytotoxicity effect was conducted using MTT assay whereas BD CycleTestTM Plus DNA Kit assay was carried out to determine the effect of extracts on cell cycle of SK-UT-1 cells. Their effect on cell cycle-related (p53 and Cyclin B1) and inflammation-related (PARP-1 and Iκβα) proteins expression were assessed using Western blot. The presence of apoptotic cell morphologies and their population was determined using double staining assays (AOPI and Annexin V FITC/PI). Real-time reverse transcription polymerase chain reaction (qRT-PCR) was implemented to assess the effect of O. stamineus extracts on apoptotic-related (BAX and BCL-2) and growth factor-related (VEGF-A, TGF-β1 and PCNA) genes expression. The results showed that OSW and OSEW have moderate and no cytotoxic effect against SK-UT-1 cells with IC50 values 18 μg/mL and 34 μg/mL, respectively. Both extracts showed no cytotoxic effects towards normal fibroblast (NIH/3T3) cells with IC50 exceed 100 μg/mL. The extracts were also found to induce cell cycle arrest at G0/G1 by OSW and G0/G1 and G2/M by OSEW by regulating the Cyclin B1 protein expression in a p53-independent manner. AO/PI staining assay showed the presence of apoptotic cell morphologies and their population in Annexin V-FITC/PI assay were increased in dose-dependent manner. qRT-PCR analysis using relative quantitative method revealed the up regulation of BAX by OSEW alone and downregulation of BCL-2 by both extracts. Western blotting revealed the presence of PARP-1 cleavage and the increase of Iκβα protein expression by both extracts. Apart from that, VEGF A, TGF-β1 and PCNA genes expression were found to be downregulated by both extracts. From this study, it can be concluded that the O. stamineus extracts possess moderate cytotoxic, anti-proliferative and anti-inflammatory effect through regulation at biochemical level expression. It also induced apoptosis in uterine leiomyosarcoma cells (SK-UT-1) in a dose-dependent manner through intrinsic pathway. |
| title | 2018_Cytotoxic Evaluation of Orthosiphon Stamineus Benth Extracts and Their Molecular Mechanism of Apoptosis on Uterine Leiomyosarcoma (SK-UT-1) Cells |
| title_full | 2018_Cytotoxic Evaluation of Orthosiphon Stamineus Benth Extracts and Their Molecular Mechanism of Apoptosis on Uterine Leiomyosarcoma (SK-UT-1) Cells |
| title_fullStr | 2018_Cytotoxic Evaluation of Orthosiphon Stamineus Benth Extracts and Their Molecular Mechanism of Apoptosis on Uterine Leiomyosarcoma (SK-UT-1) Cells |
| title_full_unstemmed | 2018_Cytotoxic Evaluation of Orthosiphon Stamineus Benth Extracts and Their Molecular Mechanism of Apoptosis on Uterine Leiomyosarcoma (SK-UT-1) Cells |
| title_short | 2018_Cytotoxic Evaluation of Orthosiphon Stamineus Benth Extracts and Their Molecular Mechanism of Apoptosis on Uterine Leiomyosarcoma (SK-UT-1) Cells |
| title_sort | 2018_cytotoxic evaluation of orthosiphon stamineus benth extracts and their molecular mechanism of apoptosis on uterine leiomyosarcoma (sk-ut-1) cells |