{"title":"山奈酚靶向全局表观遗传修饰因子阻碍HeLa细胞的生长和迁移能力","authors":"Nazia Afroze, Shafiul Haque, Arif Hussain","doi":"10.1111/jcmm.70498","DOIUrl":null,"url":null,"abstract":"<p>Dietary bioactive agents can curb tumour progression through chromatin alterations. Thus, this study attempts to evaluate the influence of kaempferol on epigenome modification in HeLa cells. Biochemical analysis for global DNA methylation-LINE 1, DNMTs (DNA methyltransferases), HAT (histone acetyl transferase), HDACs (histone deacetylases) and HMTs (histone methyltransferases) were examined with their transcript level expression through qPCR. Also, H3 and H4 histone modification marks were quantitated by an ELISA-based assay. Moreover, qPCR and protein profiler were performed to analyse the expression of migratory genes at both mRNA and protein levels, respectively, that was further substantiated through colony formation, invasion, and scratch wound assays. Finally, DNA methyl-sequencing was performed to analyse the promoter methylation of TSGs (tumour suppressor genes) and corroborated by analysing selected TSGs' expression. Kaempferol treatment did not alter the global DNA methylation-LINE 1 compared to untreated control, however, it reduced the expression and biochemical activities of DNMT and HDAC, which can be linked to their hypermethylation by kaempferol exposure. Concordant with the reduced expression of HMTs, HATs and other epi-enzymes, various histone H3 and H4 marks were also observed to be modulated. Kaempferol exposure led to promoter hypomethylation of various TSGs (such as <i>WIF1, RUNX1, RARβ, SOX1</i>), which subsequently led to enhanced expression at the mRNA level, which corresponds to their reactivation. Molecular studies were consistent with cell-based studies, which demonstrated a strong growth inhibitory and anti-migratory effect of kaempferol. This research helps to understand the probable mechanism used by kaempferol as a potential epigenetic modifier.</p>","PeriodicalId":101321,"journal":{"name":"JOURNAL OF CELLULAR AND MOLECULAR MEDICINE","volume":"29 7","pages":""},"PeriodicalIF":5.3000,"publicationDate":"2025-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jcmm.70498","citationCount":"0","resultStr":"{\"title\":\"Kaempferol Targets Global Epigenetic Modifiers to Impedes Growth and Migratory Ability of HeLa Cells\",\"authors\":\"Nazia Afroze, Shafiul Haque, Arif Hussain\",\"doi\":\"10.1111/jcmm.70498\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Dietary bioactive agents can curb tumour progression through chromatin alterations. Thus, this study attempts to evaluate the influence of kaempferol on epigenome modification in HeLa cells. Biochemical analysis for global DNA methylation-LINE 1, DNMTs (DNA methyltransferases), HAT (histone acetyl transferase), HDACs (histone deacetylases) and HMTs (histone methyltransferases) were examined with their transcript level expression through qPCR. Also, H3 and H4 histone modification marks were quantitated by an ELISA-based assay. Moreover, qPCR and protein profiler were performed to analyse the expression of migratory genes at both mRNA and protein levels, respectively, that was further substantiated through colony formation, invasion, and scratch wound assays. Finally, DNA methyl-sequencing was performed to analyse the promoter methylation of TSGs (tumour suppressor genes) and corroborated by analysing selected TSGs' expression. Kaempferol treatment did not alter the global DNA methylation-LINE 1 compared to untreated control, however, it reduced the expression and biochemical activities of DNMT and HDAC, which can be linked to their hypermethylation by kaempferol exposure. Concordant with the reduced expression of HMTs, HATs and other epi-enzymes, various histone H3 and H4 marks were also observed to be modulated. Kaempferol exposure led to promoter hypomethylation of various TSGs (such as <i>WIF1, RUNX1, RARβ, SOX1</i>), which subsequently led to enhanced expression at the mRNA level, which corresponds to their reactivation. Molecular studies were consistent with cell-based studies, which demonstrated a strong growth inhibitory and anti-migratory effect of kaempferol. 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引用次数: 0
摘要
膳食生物活性制剂可以通过改变染色质来抑制肿瘤的发展。因此,本研究试图评估山奈酚对HeLa细胞表观基因组修饰的影响。通过qPCR检测全球DNA甲基化- line 1、DNA甲基转移酶(dnmt)、组蛋白乙酰转移酶(HAT)、组蛋白去乙酰化酶(hdac)和组蛋白甲基转移酶(hmt)的生化分析及其转录本水平表达。此外,H3和H4组蛋白修饰标记通过elisa法定量。此外,利用qPCR和蛋白谱仪分别分析了迁移基因在mRNA和蛋白水平上的表达,并通过集落形成、入侵和抓伤实验进一步证实了这一点。最后,通过DNA甲基化测序分析TSGs(肿瘤抑制基因)的启动子甲基化,并通过分析选定的TSGs的表达来证实。与未处理的对照组相比,山奈酚处理没有改变总体DNA甲基化- line - 1,然而,它降低了DNMT和HDAC的表达和生化活性,这可能与山奈酚暴露导致的它们的高甲基化有关。与hmt、HATs等外表酶的表达减少一致,组蛋白H3、H4的各种标记也被调节。山奈酚暴露导致各种tsg(如WIF1、RUNX1、RARβ、SOX1)的启动子低甲基化,随后导致mRNA水平上的表达增强,这与它们的再激活相对应。分子研究结果与细胞研究结果一致,表明山奈酚具有较强的生长抑制和抗迁移作用。这项研究有助于了解山奈酚作为潜在表观遗传修饰物的可能机制。
Kaempferol Targets Global Epigenetic Modifiers to Impedes Growth and Migratory Ability of HeLa Cells
Dietary bioactive agents can curb tumour progression through chromatin alterations. Thus, this study attempts to evaluate the influence of kaempferol on epigenome modification in HeLa cells. Biochemical analysis for global DNA methylation-LINE 1, DNMTs (DNA methyltransferases), HAT (histone acetyl transferase), HDACs (histone deacetylases) and HMTs (histone methyltransferases) were examined with their transcript level expression through qPCR. Also, H3 and H4 histone modification marks were quantitated by an ELISA-based assay. Moreover, qPCR and protein profiler were performed to analyse the expression of migratory genes at both mRNA and protein levels, respectively, that was further substantiated through colony formation, invasion, and scratch wound assays. Finally, DNA methyl-sequencing was performed to analyse the promoter methylation of TSGs (tumour suppressor genes) and corroborated by analysing selected TSGs' expression. Kaempferol treatment did not alter the global DNA methylation-LINE 1 compared to untreated control, however, it reduced the expression and biochemical activities of DNMT and HDAC, which can be linked to their hypermethylation by kaempferol exposure. Concordant with the reduced expression of HMTs, HATs and other epi-enzymes, various histone H3 and H4 marks were also observed to be modulated. Kaempferol exposure led to promoter hypomethylation of various TSGs (such as WIF1, RUNX1, RARβ, SOX1), which subsequently led to enhanced expression at the mRNA level, which corresponds to their reactivation. Molecular studies were consistent with cell-based studies, which demonstrated a strong growth inhibitory and anti-migratory effect of kaempferol. This research helps to understand the probable mechanism used by kaempferol as a potential epigenetic modifier.
期刊介绍:
The Journal of Cellular and Molecular Medicine serves as a bridge between physiology and cellular medicine, as well as molecular biology and molecular therapeutics. With a 20-year history, the journal adopts an interdisciplinary approach to showcase innovative discoveries.
It publishes research aimed at advancing the collective understanding of the cellular and molecular mechanisms underlying diseases. The journal emphasizes translational studies that translate this knowledge into therapeutic strategies. Being fully open access, the journal is accessible to all readers.