2022_Elucidating the Therapeutic Impact of Thymoquinone on C-Myc Oncogene in Acute Myeloid Leukemic Cell Line
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| date | 2022-08-30 |
| format | General Document |
| id | 15751 |
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| internalnotes | Sila masukkan subject wajib Dissertations, Academic. Terima kasih... |
| originalfilename | ELUCIDATING THE THERAPEUTIC IMPACT OF THYMOQUINONE ON C-MYC ONCOGENE IN ACUTE MYELOID LEUKEMIC CELL LINE (PHD_2022).pdf |
| person | Belal Bassam Almajali |
| recordtype | oai_dc |
| resourceurl | https://intelek.unisza.edu.my/intelek/pages/view.php?ref=15751 |
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| spelling | 15751 https://intelek.unisza.edu.my/intelek/pages/view.php?ref=15751 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 Health Sciences English application/pdf 1.5 265 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) Copyright©PWB2025 Thymoquinone—Pharmacology. Thymoquinone—Pharmacology ELUCIDATING THE THERAPEUTIC IMPACT OF THYMOQUINONE ON C-MYC ONCOGENE IN ACUTE MYELOID LEUKEMIC CELL LINE (PHD_2022).pdf 2022_Elucidating the Therapeutic Impact of Thymoquinone on C-Myc Oncogene in Acute Myeloid Leukemic Cell Line Belal Bassam Almajali 2022-08-30 Acute myeloid leukemia (AML) is a blood cancer characterized by infiltration of bone marrow (BM) with malignant myeloblasts due to abnormal cytogenetic events. Overexpression of c-Myc plays an essential role in leukemogenesis and drug resistance, making c-Myc an attractive target for cancer therapy. It is impossible to target c-Myc directly, and instead, can be targeted indirectly through c-Myc-mediated signaling pathways. To date, AML patients are treated by intensive chemotherapy. However, besides the high cost of chemotherapy and its side effects, 50% of cases developed resistance. Therefore, an alternative and low side effect treatment option is vitally needed. Thymoquinone (TQ) is an essential bioactive ingredient in Nigella sativa and has demonstrated potential anticancer properties. Several studies have shown that TQ has inhibited cell proliferation and enhanced apoptosis in myeloid leukemia cells, breast cancer cells, and multiple myeloma. However, the effect mechanism of TQ on AML cells still not fully understood. The present study was performed to elucidate the effect mechanisms of TQ on HL60 AML cells. Cell viability and cytotoxic assays, followed by apoptosis assay were performed using MTT and FITC Annexin, respectively. Other molecular parameters were also performed before and after treatment with TQ including gene expression profiling using RNA sequencing followed by reverse transcriptase polymerase chain reaction (RT-qPCR) to validate the expression of c-Myc and genes involved in JAK/STAT and PI3K/AKT/mTOR pathways. In addition, Jess assay analysis was also performed to determine the effects of TQ on JAK/STAT, PI3K/AKT, and c-Myc protein expression. Two-way ANOVA, Kruskal–Wallis and Mann-Whitney tests were conducted for statistical analysis using GraphPad Prism 8.4.3, and P<0.05 was considered as statistically significant. The finding of MTT assay has revealed that 2µM was the 50% inhibitory concentration (IC50) of TQ on HL60 leukemia cells. Annexin-FITC has showed that TQ significantly (P<0.001) increased the apoptotic cells' percentage in dose and time-dependent manner. The cell cycle analysis using Guava® reagent has showed notably induced cycle arrest at G0-G1 phase (P<0.001) after treatment of HL60 cells with 1, 2, and 3-µM of TQ. The results of gene expression profiling have revealed the change of expression level for 114 genes in HL60 TQ treated cells either significantly down or up-regulation (p-adj<0.05, fold change > |1|). Then, the results were further analyzed using DAVID online free software which revealed that most of these genes are related to apoptosis and proliferation process. The results of RT-qPCR showed a marked downregulation in JAK2, STAT3, STAT5a, STAT5b, c-Myc, PI3K, AKT, and mTOR. The findings have also demonstrated that TQ inhibited JAK/STAT and PI3K/AKT signaling, leading to inhibition of c-Myc protein expression. The findings of this work support the hypothesis that TQ inhibited c-Myc proto-oncogene in HL60 cells through inhibition of c-Myc upstream regulators JAK/STAT and PI3K/AKT signaling pathways and TQ could be a potential treatment candidate for AML patients with c-Myc oncogene. Dissertations, Academic Sila masukkan subject wajib Dissertations, Academic. Terima kasih... Thymoquinone And Leukemia Treatment C-Myc Oncogene Suppression Thesis |
| spellingShingle | 2022_Elucidating the Therapeutic Impact of Thymoquinone on C-Myc Oncogene in Acute Myeloid Leukemic Cell Line |
| state | Terengganu |
| subject | Thymoquinone—Pharmacology Dissertations, Academic |
| summary | Acute myeloid leukemia (AML) is a blood cancer characterized by infiltration of bone marrow (BM) with malignant myeloblasts due to abnormal cytogenetic events. Overexpression of c-Myc plays an essential role in leukemogenesis and drug resistance, making c-Myc an attractive target for cancer therapy. It is impossible to target c-Myc directly, and instead, can be targeted indirectly through c-Myc-mediated signaling pathways. To date, AML patients are treated by intensive chemotherapy. However, besides the high cost of chemotherapy and its side effects, 50% of cases developed resistance. Therefore, an alternative and low side effect treatment option is vitally needed. Thymoquinone (TQ) is an essential bioactive ingredient in Nigella sativa and has demonstrated potential anticancer properties. Several studies have shown that TQ has inhibited cell proliferation and enhanced apoptosis in myeloid leukemia cells, breast cancer cells, and multiple myeloma. However, the effect mechanism of TQ on AML cells still not fully understood. The present study was performed to elucidate the effect mechanisms of TQ on HL60 AML cells. Cell viability and cytotoxic assays, followed by apoptosis assay were performed using MTT and FITC Annexin, respectively. Other molecular parameters were also performed before and after treatment with TQ including gene expression profiling using RNA sequencing followed by reverse transcriptase polymerase chain reaction (RT-qPCR) to validate the expression of c-Myc and genes involved in JAK/STAT and PI3K/AKT/mTOR pathways. In addition, Jess assay analysis was also performed to determine the effects of TQ on JAK/STAT, PI3K/AKT, and c-Myc protein expression. Two-way ANOVA, Kruskal–Wallis and Mann-Whitney tests were conducted for statistical analysis using GraphPad Prism 8.4.3, and P<0.05 was considered as statistically significant. The finding of MTT assay has revealed that 2µM was the 50% inhibitory concentration (IC50) of TQ on HL60 leukemia cells. Annexin-FITC has showed that TQ significantly (P<0.001) increased the apoptotic cells' percentage in dose and time-dependent manner. The cell cycle analysis using Guava® reagent has showed notably induced cycle arrest at G0-G1 phase (P<0.001) after treatment of HL60 cells with 1, 2, and 3-µM of TQ. The results of gene expression profiling have revealed the change of expression level for 114 genes in HL60 TQ treated cells either significantly down or up-regulation (p-adj<0.05, fold change > |1|). Then, the results were further analyzed using DAVID online free software which revealed that most of these genes are related to apoptosis and proliferation process. The results of RT-qPCR showed a marked downregulation in JAK2, STAT3, STAT5a, STAT5b, c-Myc, PI3K, AKT, and mTOR. The findings have also demonstrated that TQ inhibited JAK/STAT and PI3K/AKT signaling, leading to inhibition of c-Myc protein expression. The findings of this work support the hypothesis that TQ inhibited c-Myc proto-oncogene in HL60 cells through inhibition of c-Myc upstream regulators JAK/STAT and PI3K/AKT signaling pathways and TQ could be a potential treatment candidate for AML patients with c-Myc oncogene. |
| title | 2022_Elucidating the Therapeutic Impact of Thymoquinone on C-Myc Oncogene in Acute Myeloid Leukemic Cell Line |
| title_full | 2022_Elucidating the Therapeutic Impact of Thymoquinone on C-Myc Oncogene in Acute Myeloid Leukemic Cell Line |
| title_fullStr | 2022_Elucidating the Therapeutic Impact of Thymoquinone on C-Myc Oncogene in Acute Myeloid Leukemic Cell Line |
| title_full_unstemmed | 2022_Elucidating the Therapeutic Impact of Thymoquinone on C-Myc Oncogene in Acute Myeloid Leukemic Cell Line |
| title_short | 2022_Elucidating the Therapeutic Impact of Thymoquinone on C-Myc Oncogene in Acute Myeloid Leukemic Cell Line |
| title_sort | 2022_elucidating the therapeutic impact of thymoquinone on c-myc oncogene in acute myeloid leukemic cell line |