Thanigaivelan Kanagasabai, Mariam Hawaz, Kayla Ellis, Orlyne Fah, Helana Mikhaeil, Philip Nguyen, Nathalie Tombo, Anil Shanker, Chethan Sampath, Zaid H Khoury, James Cade, Alexys Ferguson, Pandu Gangula
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The critical parameters of cellular metabolism, including extracellular acidification rate (ECAR) and mitochondrial oxidative phosphorylation based on the oxygen consumption rate of cancer cells were assessed using Seahorse assay. Western blotting was performed to assess the proteins that are associated with proliferation (Survivin, IL-6) and cancer stem cell function (Oct4, Sox2) in cell lysates prepared from control and etoposide treated groups. Statistical analysis was performed using One-way ANOVA with Dunnett's multiple comparisons test.</p><p><strong>Results: </strong>The protein expression of TOP2A was significantly (P<0.05) inhibited by etoposide. Additionally, TOP2A inhibition decreased the mitochondrial respiratory parameters including basal respiration, maximal respiration and ATP production. However, TOP2A inhibition has no impact on glycolytic function. Moreover, the proliferative marker survivin and IL-6 showed a significant (P<0.05) decrease after TOP2A inhibition. Conversely, the protein expression of cancer stem cell markers Oct-4 and Sox 2 were not altered.</p><p><strong>Conclusion: </strong>These results indicate that inhibition of TOP2A is more efficacious by decreasing the mitochondrial metabolic reprogramming and thereby downregulating the key anti-apoptotic and pro-survival mediators. Thus, TOP2A represents an ideal therapeutic target and offers a potential treatment strategy for OSCC.</p>","PeriodicalId":93742,"journal":{"name":"Dental research and oral health","volume":"7 2","pages":"58-65"},"PeriodicalIF":0.0000,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11218736/pdf/","citationCount":"0","resultStr":"{\"title\":\"Effects of Topoisomerase II alpha Inhibition on Oral Cancer Cell Metabolism and Cancer Stem Cell Function.\",\"authors\":\"Thanigaivelan Kanagasabai, Mariam Hawaz, Kayla Ellis, Orlyne Fah, Helana Mikhaeil, Philip Nguyen, Nathalie Tombo, Anil Shanker, Chethan Sampath, Zaid H Khoury, James Cade, Alexys Ferguson, Pandu Gangula\",\"doi\":\"10.26502/droh.0076\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Topoisomerase IIα (TOP2A), is an enzyme involved in DNA replication, transcription, recombination, and chromatin remodeling and is found in a variety of cancers. However, the role of TOP2A regulation in oral cancer progression is not fully explained. We investigated the effect of TOP2A inhibition on cell survival, metabolism, and cancer stem cell self-renewal function in oral cancer cells.</p><p><strong>Methods: </strong>Oral carcinoma cell line SCC25 was cultured in complete DMEM/F12 media and treated with 5μM of Etoposide (Topoisomerase II inhibitor) for 48h. The critical parameters of cellular metabolism, including extracellular acidification rate (ECAR) and mitochondrial oxidative phosphorylation based on the oxygen consumption rate of cancer cells were assessed using Seahorse assay. Western blotting was performed to assess the proteins that are associated with proliferation (Survivin, IL-6) and cancer stem cell function (Oct4, Sox2) in cell lysates prepared from control and etoposide treated groups. Statistical analysis was performed using One-way ANOVA with Dunnett's multiple comparisons test.</p><p><strong>Results: </strong>The protein expression of TOP2A was significantly (P<0.05) inhibited by etoposide. Additionally, TOP2A inhibition decreased the mitochondrial respiratory parameters including basal respiration, maximal respiration and ATP production. However, TOP2A inhibition has no impact on glycolytic function. Moreover, the proliferative marker survivin and IL-6 showed a significant (P<0.05) decrease after TOP2A inhibition. Conversely, the protein expression of cancer stem cell markers Oct-4 and Sox 2 were not altered.</p><p><strong>Conclusion: </strong>These results indicate that inhibition of TOP2A is more efficacious by decreasing the mitochondrial metabolic reprogramming and thereby downregulating the key anti-apoptotic and pro-survival mediators. 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引用次数: 0
摘要
背景：拓扑异构酶 IIα (TOP2A)是一种参与 DNA 复制、转录、重组和染色质重塑的酶，存在于多种癌症中。然而，TOP2A 在口腔癌进展过程中的调控作用尚未得到充分解释。我们研究了抑制 TOP2A 对口腔癌细胞存活、代谢和癌症干细胞自我更新功能的影响：方法：口腔癌细胞株 SCC25 在完全 DMEM/F12 培养基中培养，并用 5μM 的依托泊苷（拓扑异构酶 II 抑制剂）处理 48 小时。使用海马测定法评估细胞代谢的关键参数，包括细胞外酸化率（ECAR）和基于癌细胞耗氧率的线粒体氧化磷酸化。对对照组和依托泊苷处理组细胞裂解物中与增殖（Survivin、IL-6）和癌症干细胞功能（Oct4、Sox2）相关的蛋白质进行了 Western 印迹评估。统计分析采用单因素方差分析和邓尼特多重比较检验：结果：TOP2A 的蛋白表达明显减少（PConclusion：这些结果表明，抑制 TOP2A 可减少线粒体代谢重编程，从而下调关键的抗凋亡和促生存介质，因此具有更好的疗效。因此，TOP2A是一个理想的治疗靶点，为OSCC提供了一种潜在的治疗策略。
Effects of Topoisomerase II alpha Inhibition on Oral Cancer Cell Metabolism and Cancer Stem Cell Function.
Background: Topoisomerase IIα (TOP2A), is an enzyme involved in DNA replication, transcription, recombination, and chromatin remodeling and is found in a variety of cancers. However, the role of TOP2A regulation in oral cancer progression is not fully explained. We investigated the effect of TOP2A inhibition on cell survival, metabolism, and cancer stem cell self-renewal function in oral cancer cells.
Methods: Oral carcinoma cell line SCC25 was cultured in complete DMEM/F12 media and treated with 5μM of Etoposide (Topoisomerase II inhibitor) for 48h. The critical parameters of cellular metabolism, including extracellular acidification rate (ECAR) and mitochondrial oxidative phosphorylation based on the oxygen consumption rate of cancer cells were assessed using Seahorse assay. Western blotting was performed to assess the proteins that are associated with proliferation (Survivin, IL-6) and cancer stem cell function (Oct4, Sox2) in cell lysates prepared from control and etoposide treated groups. Statistical analysis was performed using One-way ANOVA with Dunnett's multiple comparisons test.
Results: The protein expression of TOP2A was significantly (P<0.05) inhibited by etoposide. Additionally, TOP2A inhibition decreased the mitochondrial respiratory parameters including basal respiration, maximal respiration and ATP production. However, TOP2A inhibition has no impact on glycolytic function. Moreover, the proliferative marker survivin and IL-6 showed a significant (P<0.05) decrease after TOP2A inhibition. Conversely, the protein expression of cancer stem cell markers Oct-4 and Sox 2 were not altered.
Conclusion: These results indicate that inhibition of TOP2A is more efficacious by decreasing the mitochondrial metabolic reprogramming and thereby downregulating the key anti-apoptotic and pro-survival mediators. Thus, TOP2A represents an ideal therapeutic target and offers a potential treatment strategy for OSCC.
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