2018_Investigation of Chemical Profile and Apoptosis Induction by Melastoma Malabathricum L. Extracts Against HEPG2 Cell Lines
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| date | 2019-02-19 |
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| id | 15484 |
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| internalnotes | Sila masukkan subject wajib Dissertations, Academic. Terima kasih... |
| originalfilename | INVESTIGATION OF CHEMICAL PROFILE AND APOPTOSIS INDUCTION BY MELASTOMA MALABATHRICUM L. EXTRACTS AGAINST HEPG2 CELL LINES (MASTER_2018).pdf |
| person | Wan Siti Amirah Binti Wan Mohd Azemin |
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| resourceurl | https://intelek.unisza.edu.my/intelek/pages/view.php?ref=15484 |
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| spelling | 15484 https://intelek.unisza.edu.my/intelek/pages/view.php?ref=15484 https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection3 General Document Library Staff (Top Management) Library Staff (Management) Library Staff (Support) Terengganu Faculty of Medicine 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) 2019-02-19 236 INVESTIGATION OF CHEMICAL PROFILE AND APOPTOSIS INDUCTION BY MELASTOMA MALABATHRICUM L. EXTRACTS AGAINST HEPG2 CELL LINES (MASTER_2018).pdf 2018_Investigation of Chemical Profile and Apoptosis Induction by Melastoma Malabathricum L. Extracts Against HEPG2 Cell Lines Copyright©PWB2025 Wan Siti Amirah Binti Wan Mohd Azemin Melastoma malabathricum—Chemical composition Hepatocellular carcinoma is third leading cause of cancer-related deaths in the world. This cancer is detected at an advanced stage and the available drugs often cause side effects. Thus, programmed cell death (apoptosis) of the cancer cells is crucial for helping the human immune system to stop this abnormal growth which is an extremely promising approach for anticancer therapy. Melastoma malabathricum (MM) is a medicinal plant that grows wildly throughout Malaysia. This plant has great potential as anticancer agent as shown by previous studies on various cancer cell lines. This study has been conducted in two parts to further elucidate this. The first part was to standardize the M. malabathricum plant from seven different localities and the second part investigated the potential of MM extracts as anticancer therapeutic through assessment of apoptotic induction in HepG2 cell lines. i) Morphological, and quality control techniques based on gravimetric analysis, preliminary chemical screening, high performance thin layer chromatographic analysis as well as Fourier Transform Infrared Spectroscopy (FTIR) metabolite fingerprinting were applied to the MM samples from different locations. ii) The mode of cell death caused by methanol and aqueous extracts against HepG2 cell lines was examined by cytotoxic assay, acridine orange (AO)/propidium iodide (PI) double-staining and annexinV-FITC/PI staining techniques. The distribution of cell cycle was assessed using flow cytometry technique. Apoptotic related proteins expression namely phosphorylated Bcl-2, tumour suppressor p53 and PARP-1 cleavage proteins were determined using immunoblotting technique. i) The authenticity of the plant was established based on quality control methods which showed that the morphological characteristics, gravimetric results and phytochemical content were similar to that reported in herbal monographs and standard references. HPTLC screening showed that either flavonoid standard marker, quercetin or triterpenic acid marker, ursolic acid offered a fast profiling method whereas the FTIR metabolite fingerprinting using LDA analysis on six principal components of the FTIR data showed good classification of the samples according to locations. ii) The cytotoxicity assays revealed methanol extract possessed the highest toxicity value (IC50=1.4µg/mL) compared to other extracts with aqueous extract being second best (IC50 = 10µg/mL). Aqueous extract showed acute and rapid responses by rupturing the membrane immediately with 31.3% and 2.9% of HepG2 cells in early apoptosis and 51.3% and 8.8% in late apoptosis as shown by AO/PI staining and Annexin V assays, respectively. The percentages of methanol extract treated HepG2 cells at late apoptosis were 30.3% and 3.8% as shown by AO/PI staining and Annexin V assays. Aqueous and methanol extracts significantly arrested cell progression at the G2/M phase where the ratio of the G2/M phase population significantly increased from 21.9% (untreated control) to 23.9% and 23.8% (24 hours), respectively. Besides, the percentages of HepG2 cells were increased at Sub Go/G1 phase from 3.8% (untreated control) to 11.7% and 12% (24 hours) for aqueous and methanol extracts, respectively, which indicated cells with DNA damage and undergoing apoptosis. Both extracts showed increased expression of key apoptotic proteins such as phosphorylated Bcl-2, p53, and PARP-1 protein compared to negative control. The research demonstrates that aqueous extract induces apoptosis event in rapid response compared to methanol extract. MM extracts significantly induced apoptosis and G2/M -phase checkpoint arrest. Thus, Melastoma malabathricum extract can potentially be developed as an anticancer therapeutic especially against hepatic cancer. Dissertations, Academic Sila masukkan subject wajib Dissertations, Academic. Terima kasih... Melastoma Malabathricum Extracts Chemical Profiling Of Medicinal Plants HepG2 Liver Cancer Cells Thesis |
| spellingShingle | 2018_Investigation of Chemical Profile and Apoptosis Induction by Melastoma Malabathricum L. Extracts Against HEPG2 Cell Lines |
| state | Terengganu |
| subject | Melastoma malabathricum—Chemical composition Dissertations, Academic |
| summary | Hepatocellular carcinoma is third leading cause of cancer-related deaths in the world. This cancer is detected at an advanced stage and the available drugs often cause side effects. Thus, programmed cell death (apoptosis) of the cancer cells is crucial for helping the human immune system to stop this abnormal growth which is an extremely promising approach for anticancer therapy. Melastoma malabathricum (MM) is a medicinal plant that grows wildly throughout Malaysia. This plant has great potential as anticancer agent as shown by previous studies on various cancer cell lines. This study has been conducted in two parts to further elucidate this. The first part was to standardize the M. malabathricum plant from seven different localities and the second part investigated the potential of MM extracts as anticancer therapeutic through assessment of apoptotic induction in HepG2 cell lines. i) Morphological, and quality control techniques based on gravimetric analysis, preliminary chemical screening, high performance thin layer chromatographic analysis as well as Fourier Transform Infrared Spectroscopy (FTIR) metabolite fingerprinting were applied to the MM samples from different locations. ii) The mode of cell death caused by methanol and aqueous extracts against HepG2 cell lines was examined by cytotoxic assay, acridine orange (AO)/propidium iodide (PI) double-staining and annexinV-FITC/PI staining techniques. The distribution of cell cycle was assessed using flow cytometry technique. Apoptotic related proteins expression namely phosphorylated Bcl-2, tumour suppressor p53 and PARP-1 cleavage proteins were determined using immunoblotting technique. i) The authenticity of the plant was established based on quality control methods which showed that the morphological characteristics, gravimetric results and phytochemical content were similar to that reported in herbal monographs and standard references. HPTLC screening showed that either flavonoid standard marker, quercetin or triterpenic acid marker, ursolic acid offered a fast profiling method whereas the FTIR metabolite fingerprinting using LDA analysis on six principal components of the FTIR data showed good classification of the samples according to locations. ii) The cytotoxicity assays revealed methanol extract possessed the highest toxicity value (IC50=1.4µg/mL) compared to other extracts with aqueous extract being second best (IC50 = 10µg/mL). Aqueous extract showed acute and rapid responses by rupturing the membrane immediately with 31.3% and 2.9% of HepG2 cells in early apoptosis and 51.3% and 8.8% in late apoptosis as shown by AO/PI staining and Annexin V assays, respectively. The percentages of methanol extract treated HepG2 cells at late apoptosis were 30.3% and 3.8% as shown by AO/PI staining and Annexin V assays. Aqueous and methanol extracts significantly arrested cell progression at the G2/M phase where the ratio of the G2/M phase population significantly increased from 21.9% (untreated control) to 23.9% and 23.8% (24 hours), respectively. Besides, the percentages of HepG2 cells were increased at Sub Go/G1 phase from 3.8% (untreated control) to 11.7% and 12% (24 hours) for aqueous and methanol extracts, respectively, which indicated cells with DNA damage and undergoing apoptosis. Both extracts showed increased expression of key apoptotic proteins such as phosphorylated Bcl-2, p53, and PARP-1 protein compared to negative control. The research demonstrates that aqueous extract induces apoptosis event in rapid response compared to methanol extract. MM extracts significantly induced apoptosis and G2/M -phase checkpoint arrest. Thus, Melastoma malabathricum extract can potentially be developed as an anticancer therapeutic especially against hepatic cancer. |
| title | 2018_Investigation of Chemical Profile and Apoptosis Induction by Melastoma Malabathricum L. Extracts Against HEPG2 Cell Lines |
| title_full | 2018_Investigation of Chemical Profile and Apoptosis Induction by Melastoma Malabathricum L. Extracts Against HEPG2 Cell Lines |
| title_fullStr | 2018_Investigation of Chemical Profile and Apoptosis Induction by Melastoma Malabathricum L. Extracts Against HEPG2 Cell Lines |
| title_full_unstemmed | 2018_Investigation of Chemical Profile and Apoptosis Induction by Melastoma Malabathricum L. Extracts Against HEPG2 Cell Lines |
| title_short | 2018_Investigation of Chemical Profile and Apoptosis Induction by Melastoma Malabathricum L. Extracts Against HEPG2 Cell Lines |
| title_sort | 2018_investigation of chemical profile and apoptosis induction by melastoma malabathricum l. extracts against hepg2 cell lines |