Keyhan Ebrahimi, Ramin Bagheri, Hamid Gholamhosseinian, Mohammad Reza Keramati, Houshang Rafatpanah, Mehrdad Iranshahi, Fatemeh B Rassouli
{"title":"通过与 CDK6 的相互作用,伞形酮改善电离辐射对成人 T 细胞白血病/淋巴瘤细胞的抗增殖作用;一项体外和硅学研究。","authors":"Keyhan Ebrahimi, Ramin Bagheri, Hamid Gholamhosseinian, Mohammad Reza Keramati, Houshang Rafatpanah, Mehrdad Iranshahi, Fatemeh B Rassouli","doi":"10.1177/03946320241287873","DOIUrl":null,"url":null,"abstract":"<p><p>Adult T-cell leukemia/lymphoma (ATL) is an aggressive malignancy with poor survival rates. The efficacy of radiotherapy in ATL needs enhancement with radiosensitizing agents. This study investigated whether umbelliprenin (UMB) could improve the therapeutic effects of ionizing radiation (IR) in ATL cells. UMB, a naturally occurring prenylated coumarin, exhibits anticancer properties and has shown synergistic effects when combined with chemotherapeutic drugs. Despite this promising profile, there is a notable lack of research on its potential combinatorial effects with IR, particularly for ATL treatment. UMB was extracted from <i>Ferula persica</i> using thin layer chromatography. MT-2 cells were treated with UMB alone and in combination with various doses of IR, and cell proliferation was assessed via alamarBlue assay. Flow cytometry with annexin V and PI staining was conducted, and candidate gene expression was analyzed by qPCR. <i>In silico</i> analysis involved identifying pathogenic targets of ATL, constructing protein-protein interaction (PPI) networks, and evaluating CDK6 expression in MT-2 cells. Molecular docking was used to determine the interaction between UMB and CDK6. The alamarBlue assay and flow cytometry showed that pretreating ATL cells with UMB significantly (<i>p</i> < .0001) enhanced anti-proliferative effects of IR. The combination index indicated a synergistic effect between UMB and IR. qPCR revealed significant (<i>p</i> < .0001) downregulation of <i>CD44, CDK6, c-MYC</i>, and <i>cFLIPL</i>, and overexpression of <i>cFLIPS</i>. Computational analysis identified CDK6 as a hub gene in the PPI network, and CDK6 overexpression was confirmed in MT-2 cells. Molecular docking revealed a favorable binding interaction between UMB and the ATP-binding site of CDK6, with a JAMDA score of -2.131, surpassing the control selonsertib. The current study provides evidence that UMB enhances the anti-proliferative effects of IR on ATL cells, and highlights the significance of targeting CDK6 in combinatorial approaches.</p>","PeriodicalId":48647,"journal":{"name":"International Journal of Immunopathology and Pharmacology","volume":"38 ","pages":"3946320241287873"},"PeriodicalIF":3.5000,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11437583/pdf/","citationCount":"0","resultStr":"{\"title\":\"Umbelliprenin improved anti-proliferative effects of ionizing radiation on adult T-cell leukemia/lymphoma cells via interaction with CDK6; an <i>in vitro</i> and <i>in silico</i> study.\",\"authors\":\"Keyhan Ebrahimi, Ramin Bagheri, Hamid Gholamhosseinian, Mohammad Reza Keramati, Houshang Rafatpanah, Mehrdad Iranshahi, Fatemeh B Rassouli\",\"doi\":\"10.1177/03946320241287873\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Adult T-cell leukemia/lymphoma (ATL) is an aggressive malignancy with poor survival rates. The efficacy of radiotherapy in ATL needs enhancement with radiosensitizing agents. This study investigated whether umbelliprenin (UMB) could improve the therapeutic effects of ionizing radiation (IR) in ATL cells. UMB, a naturally occurring prenylated coumarin, exhibits anticancer properties and has shown synergistic effects when combined with chemotherapeutic drugs. Despite this promising profile, there is a notable lack of research on its potential combinatorial effects with IR, particularly for ATL treatment. UMB was extracted from <i>Ferula persica</i> using thin layer chromatography. MT-2 cells were treated with UMB alone and in combination with various doses of IR, and cell proliferation was assessed via alamarBlue assay. Flow cytometry with annexin V and PI staining was conducted, and candidate gene expression was analyzed by qPCR. <i>In silico</i> analysis involved identifying pathogenic targets of ATL, constructing protein-protein interaction (PPI) networks, and evaluating CDK6 expression in MT-2 cells. Molecular docking was used to determine the interaction between UMB and CDK6. The alamarBlue assay and flow cytometry showed that pretreating ATL cells with UMB significantly (<i>p</i> < .0001) enhanced anti-proliferative effects of IR. The combination index indicated a synergistic effect between UMB and IR. qPCR revealed significant (<i>p</i> < .0001) downregulation of <i>CD44, CDK6, c-MYC</i>, and <i>cFLIPL</i>, and overexpression of <i>cFLIPS</i>. Computational analysis identified CDK6 as a hub gene in the PPI network, and CDK6 overexpression was confirmed in MT-2 cells. Molecular docking revealed a favorable binding interaction between UMB and the ATP-binding site of CDK6, with a JAMDA score of -2.131, surpassing the control selonsertib. 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引用次数: 0
摘要
成人 T 细胞白血病/淋巴瘤(ATL)是一种侵袭性恶性肿瘤,存活率很低。放疗对 ATL 的疗效需要放射增敏剂来提高。本研究探讨了脐橙素(UMB)能否改善电离辐射(IR)对ATL细胞的治疗效果。UMB 是一种天然的前酰化香豆素,具有抗癌特性,并在与化疗药物联合使用时显示出协同效应。尽管UMB具有良好的抗癌特性,但关于它与IR的潜在组合效应,尤其是在ATL治疗方面的组合效应的研究却明显不足。采用薄层色谱法从阿魏中提取 UMB。用 UMB 单独或与不同剂量的红外线联合处理 MT-2 细胞,并通过氨蓝检测法评估细胞增殖情况。采用附子素 V 和 PI 染色法进行流式细胞术检测,并通过 qPCR 分析候选基因的表达。硅学分析包括确定ATL的致病靶点、构建蛋白质-蛋白质相互作用(PPI)网络以及评估MT-2细胞中CDK6的表达。分子对接用于确定 UMB 与 CDK6 之间的相互作用。钴蓝检测法和流式细胞术显示,用UMB预处理ATL细胞能显著增强IR的抗增殖作用(p < .0001)。qPCR显示CD44、CDK6、c-MYC和cFLIPL显著下调(p < .0001),而cFLIPS则过表达。计算分析发现 CDK6 是 PPI 网络中的一个枢纽基因,CDK6 在 MT-2 细胞中的过表达也得到了证实。分子对接显示,UMB与CDK6的ATP结合位点有良好的结合相互作用,其JAMDA得分为-2.131,超过了对照组selonsertib。目前的研究提供了 UMB 可增强 IR 对 ATL 细胞抗增殖作用的证据,并强调了在组合方法中靶向 CDK6 的重要性。
Umbelliprenin improved anti-proliferative effects of ionizing radiation on adult T-cell leukemia/lymphoma cells via interaction with CDK6; an in vitro and in silico study.
Adult T-cell leukemia/lymphoma (ATL) is an aggressive malignancy with poor survival rates. The efficacy of radiotherapy in ATL needs enhancement with radiosensitizing agents. This study investigated whether umbelliprenin (UMB) could improve the therapeutic effects of ionizing radiation (IR) in ATL cells. UMB, a naturally occurring prenylated coumarin, exhibits anticancer properties and has shown synergistic effects when combined with chemotherapeutic drugs. Despite this promising profile, there is a notable lack of research on its potential combinatorial effects with IR, particularly for ATL treatment. UMB was extracted from Ferula persica using thin layer chromatography. MT-2 cells were treated with UMB alone and in combination with various doses of IR, and cell proliferation was assessed via alamarBlue assay. Flow cytometry with annexin V and PI staining was conducted, and candidate gene expression was analyzed by qPCR. In silico analysis involved identifying pathogenic targets of ATL, constructing protein-protein interaction (PPI) networks, and evaluating CDK6 expression in MT-2 cells. Molecular docking was used to determine the interaction between UMB and CDK6. The alamarBlue assay and flow cytometry showed that pretreating ATL cells with UMB significantly (p < .0001) enhanced anti-proliferative effects of IR. The combination index indicated a synergistic effect between UMB and IR. qPCR revealed significant (p < .0001) downregulation of CD44, CDK6, c-MYC, and cFLIPL, and overexpression of cFLIPS. Computational analysis identified CDK6 as a hub gene in the PPI network, and CDK6 overexpression was confirmed in MT-2 cells. Molecular docking revealed a favorable binding interaction between UMB and the ATP-binding site of CDK6, with a JAMDA score of -2.131, surpassing the control selonsertib. The current study provides evidence that UMB enhances the anti-proliferative effects of IR on ATL cells, and highlights the significance of targeting CDK6 in combinatorial approaches.
期刊介绍:
International Journal of Immunopathology and Pharmacology is an Open Access peer-reviewed journal publishing original papers describing research in the fields of immunology, pathology and pharmacology. The intention is that the journal should reflect both the experimental and clinical aspects of immunology as well as advances in the understanding of the pathology and pharmacology of the immune system.