2022_Deciphering the Effect Mechanism of Thymoquinone on JAK / STAT and PI3K/AKT Signalling Negative Regulation in Myeloid Leukaemia
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| collectionurl | https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection3 |
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| country | Malaysia |
| date | 2022-10-02 |
| format | General Document |
| id | 15741 |
| institution | UniSZA |
| internalnotes | Sila masukkan subject wajib Dissertations, Academic. Terima kasih... |
| originalfilename | DECIPHERING THE EFFECT MECHANISM OF THYMOQUINONE ON JAK _STAT AND PI3K_AKT SIGNALLING NEGATIVE REGULATION IN MYELOID LEUKAEMIA (PHD_2022).pdf |
| person | Futoon Abedrabbu Falah Al-Rawashde |
| recordtype | oai_dc |
| resourceurl | https://intelek.unisza.edu.my/intelek/pages/view.php?ref=15741 |
| sourcemedia | Server storage Scanned document |
| spelling | 15741 https://intelek.unisza.edu.my/intelek/pages/view.php?ref=15741 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 1.5 298 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 DECIPHERING THE EFFECT MECHANISM OF THYMOQUINONE ON JAK _STAT AND PI3K_AKT SIGNALLING NEGATIVE REGULATION IN MYELOID LEUKAEMIA (PHD_2022).pdf 2022_Deciphering the Effect Mechanism of Thymoquinone on JAK / STAT and PI3K/AKT Signalling Negative Regulation in Myeloid Leukaemia 2022-10-02 Futoon Abedrabbu Falah Al-Rawashde Thymoquinone—Pharmacological effects Leukaemia is a heterogeneous blood malignancy caused by genetic and epigenetic alterations. Epigenetic silencing of tumour suppressor genes (TSGs) plays an essential role in leukemogenesis. Hyperactivation of JAK/STAT and PI3K/Akt/mTOR signalling is critical in the pathogenesis of acute myeloid leukaemia (AML) and chronic myeloid leukaemia (CML). SHP-1, SOCS-1, and SOCS-3 are TSGs that negatively regulate JAK/STAT signalling. Re-expression of SHP-1, SOCS-1, and SOCS-3 through demethylation represents therapeutic targets in several cancers, including leukaemia. Thymoquinone (TQ) is a major component of Nigella sativa seeds that has shown anti cancer activities. However, the anti-leukaemia activities of TQ and its effect on DNA methylation are not entirely investigated. Therefore, this study aimed to evaluate the potential of TQ to re-express SHP-1, SOCS-1, and SOCS-3 through demethylation and the consequent effect on the proliferation and apoptosis of myeloid leukaemia cells. The half maximal inhibitory concentration (IC50) of TQ in K562 and MV4-11 leukaemia cells were determined after 24, 48 and 72 hours of treatment by using WST-8 assay. The cells were then treated with the IC50 of TQ for the other analysis. Apoptosis and cell cycle progression were studied using Annexin V-FITC/PI apoptosis detection kit and fluorometric-red cell cycle assay kit, respectively, and using flow cytometry. The expression of target genes was evaluated by quantitative reverse transcription PCR (RT qPCR). Methylation of SHP-1, SOCS-1, and SOCS-3 was determined by pyrosequencing analysis. Protein and phosphorylation levels of target proteins were measured using Jess Western analysis. The molecular docking of TQ to DNMT1, DNMT3A, DNMT3B, JAK2, STAT3, and STAT5 was evaluated. The data between groups were analyzed using repeated measures ANOVA, Paired sample t-test and Wilcoxon signed-rank test using SPSS. TQ significantly downregulated DNMT1, DNMT3A, and DNMT3B, and upregulated TET2 and WT1 in K562 and MV4-11 cells. This led to a significant hypomethylation of SHP-1 and SOCS-3 in K562 cells and MV4 11 cells compared to untreated cells. SOCS-1 was unmethylated in K562 and MV4-11 cells. Hypomethylation of SHP-1 and SOCS-3 resulted in the restoration of their expression. TQ significantly downregulated BCR-ABL in K562 cells and FLT3-ITD in MV4-11 cells. TQ also induced significant downregulation of JAK2, STAT3, and STAT5 at the mRNA and protein levels in K562 and MV4-11 cells. It also binds the active sites of DNMT1, DNMT3A, DNMT3B, JAK2, STAT3, and STAT5 to inhibit their enzymatic activity. Re-expression of SHP-1 and SOCS-3 was associated with decreased phosphorylation of JAK2, STAT3, STAT5, PI3K, and Akt which resulted in growth inhibition, cell cycle arrest, and apoptosis induction in K562 and MV4-11 cells. TQ mediated anti-leukaemia effects on myeloid leukaemia cells by creating a balance in the expression of the epigenetic regulator genes, resulting in hypomethylation and re expression of SHP-1 and SOCS-3. Restoration of SHP-1 and SOCS-3 expression led to inhibition of JAK/STAT and PI3K/Akt/mTOR signalling and a consequent inhibition of cell proliferation, arrest of cell cycle, and induction of apoptosis. Dissertations, Academic Sila masukkan subject wajib Dissertations, Academic. Terima kasih... Thymoquinone And Leukemia Treatment JAK/STAT Signaling Inhibition PI3K/AKT Pathway Suppression Thesis |
| spellingShingle | 2022_Deciphering the Effect Mechanism of Thymoquinone on JAK / STAT and PI3K/AKT Signalling Negative Regulation in Myeloid Leukaemia |
| state | Terengganu |
| subject | Thymoquinone—Pharmacological effects Dissertations, Academic |
| summary | Leukaemia is a heterogeneous blood malignancy caused by genetic and epigenetic alterations. Epigenetic silencing of tumour suppressor genes (TSGs) plays an essential role in leukemogenesis. Hyperactivation of JAK/STAT and PI3K/Akt/mTOR signalling is critical in the pathogenesis of acute myeloid leukaemia (AML) and chronic myeloid leukaemia (CML). SHP-1, SOCS-1, and SOCS-3 are TSGs that negatively regulate JAK/STAT signalling. Re-expression of SHP-1, SOCS-1, and SOCS-3 through demethylation represents therapeutic targets in several cancers, including leukaemia. Thymoquinone (TQ) is a major component of Nigella sativa seeds that has shown anti cancer activities. However, the anti-leukaemia activities of TQ and its effect on DNA methylation are not entirely investigated. Therefore, this study aimed to evaluate the potential of TQ to re-express SHP-1, SOCS-1, and SOCS-3 through demethylation and the consequent effect on the proliferation and apoptosis of myeloid leukaemia cells. The half maximal inhibitory concentration (IC50) of TQ in K562 and MV4-11 leukaemia cells were determined after 24, 48 and 72 hours of treatment by using WST-8 assay. The cells were then treated with the IC50 of TQ for the other analysis. Apoptosis and cell cycle progression were studied using Annexin V-FITC/PI apoptosis detection kit and fluorometric-red cell cycle assay kit, respectively, and using flow cytometry. The expression of target genes was evaluated by quantitative reverse transcription PCR (RT qPCR). Methylation of SHP-1, SOCS-1, and SOCS-3 was determined by pyrosequencing analysis. Protein and phosphorylation levels of target proteins were measured using Jess Western analysis. The molecular docking of TQ to DNMT1, DNMT3A, DNMT3B, JAK2, STAT3, and STAT5 was evaluated. The data between groups were analyzed using repeated measures ANOVA, Paired sample t-test and Wilcoxon signed-rank test using SPSS. TQ significantly downregulated DNMT1, DNMT3A, and DNMT3B, and upregulated TET2 and WT1 in K562 and MV4-11 cells. This led to a significant hypomethylation of SHP-1 and SOCS-3 in K562 cells and MV4 11 cells compared to untreated cells. SOCS-1 was unmethylated in K562 and MV4-11 cells. Hypomethylation of SHP-1 and SOCS-3 resulted in the restoration of their expression. TQ significantly downregulated BCR-ABL in K562 cells and FLT3-ITD in MV4-11 cells. TQ also induced significant downregulation of JAK2, STAT3, and STAT5 at the mRNA and protein levels in K562 and MV4-11 cells. It also binds the active sites of DNMT1, DNMT3A, DNMT3B, JAK2, STAT3, and STAT5 to inhibit their enzymatic activity. Re-expression of SHP-1 and SOCS-3 was associated with decreased phosphorylation of JAK2, STAT3, STAT5, PI3K, and Akt which resulted in growth inhibition, cell cycle arrest, and apoptosis induction in K562 and MV4-11 cells. TQ mediated anti-leukaemia effects on myeloid leukaemia cells by creating a balance in the expression of the epigenetic regulator genes, resulting in hypomethylation and re expression of SHP-1 and SOCS-3. Restoration of SHP-1 and SOCS-3 expression led to inhibition of JAK/STAT and PI3K/Akt/mTOR signalling and a consequent inhibition of cell proliferation, arrest of cell cycle, and induction of apoptosis. |
| title | 2022_Deciphering the Effect Mechanism of Thymoquinone on JAK / STAT and PI3K/AKT Signalling Negative Regulation in Myeloid Leukaemia |
| title_full | 2022_Deciphering the Effect Mechanism of Thymoquinone on JAK / STAT and PI3K/AKT Signalling Negative Regulation in Myeloid Leukaemia |
| title_fullStr | 2022_Deciphering the Effect Mechanism of Thymoquinone on JAK / STAT and PI3K/AKT Signalling Negative Regulation in Myeloid Leukaemia |
| title_full_unstemmed | 2022_Deciphering the Effect Mechanism of Thymoquinone on JAK / STAT and PI3K/AKT Signalling Negative Regulation in Myeloid Leukaemia |
| title_short | 2022_Deciphering the Effect Mechanism of Thymoquinone on JAK / STAT and PI3K/AKT Signalling Negative Regulation in Myeloid Leukaemia |
| title_sort | 2022_deciphering the effect mechanism of thymoquinone on jak / stat and pi3k/akt signalling negative regulation in myeloid leukaemia |