| 黄俊玲,钟腾猛,黄森平,等.ESET通过 lncRNA GMDS-AS1调控肝癌细胞增殖、凋亡、糖酵解的研究[J].安徽医药,2023,27(12):2402-2407. |
| ESET通过 lncRNA GMDS-AS1调控肝癌细胞增殖、凋亡、糖酵解的研究 |
| Histone methyltransferase ESET regulates the proliferation, apoptosis and glycolysis of hepatoma cells by inhibiting lncRNA GMDS-AS1 |
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| DOI：10.3969/j.issn.1009-6469.2023.12.015 |
| 中文关键词: 肝肿瘤 甲基转移酶 ESET 长链非编码 RNA甘露糖 4，6-脱水酶反义 RNA1 增殖 凋亡 糖酵解 |
| 英文关键词: Liver neoplasms Methyltransferase ESET Long noncoding RNA mannose 4, 6-dehydrase antisense RNA1 Prolif. eration Apoptosis Glycolysis |
| 基金项目:广西壮族自治区卫生健康委自筹经费科研课题( Z-L20230906)；右江民族医学院附属医院 2019年度第一批高层次人才科研项目( Y20196303) |
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| 中文摘要: |
| 目的探究组蛋白甲基转移酶集合域分叉 1(ESET)、长链非编码 RNA甘露糖 4，6-脱水酶反义 RNA1(lncRNA GMDS-AS1)对肝癌细胞增殖、凋亡、糖酵解的调控及二者之间的潜在关系。方法收集右江民族医学院附属医院和百色市人民医院 2019年 1月至 2020年 1月期间手术切除的肝癌组织及癌旁组织 49例。脂质体法将空白组(不做任何处理)、空载体组(转染 pcDNA或 si-con)、敲减 ESET组(转染 si-ESET)、过表达 lncRNA GMDS-AS1组(转染 pcDNA-GMDS-AS1)转染至 HepG2细胞；实时荧光定量逆转录聚合酶链反应( qRT-PCR)实验、蛋白质印迹法实验检测 ESET、lncRNA GMDS-AS1、细胞增殖抗原标志物 Ki-67、葡萄糖转运蛋白 1(GLUT-1)、乳酸脱氢酶 A(LDHA)、活化胱天蛋白酶 -3(C-caspase-3)的表达；细胞计数试剂盒( CCK8)法、流式细胞术检测细胞增殖、凋亡； RNA结合蛋白免疫沉淀(RIP)实验检测 ESET与 lncRNA GMDS-AS1的关系。结果肝癌组织中 ESETmRNA和蛋白表达量为 5.18±0.94、0.69±0.14，显著高于癌旁组织的 0.99±0.33、0.23±0.07，肝癌组织 lncRNA GMDS-AS1表达量为 0.63±0.20，显著低于癌旁组织的 1.02±0.31(P<0.05)二者呈负相关性；与空载体组相比，敲减 ESET组细胞 ESET的表达明显降低，细胞增殖抗原标志物 Ki-67蛋白表达显著降低，，细胞活性显著降低， GLUT-1、LDHA的蛋白表达明显降低， C-caspase-3蛋白表达显著升高，凋亡率显著升高( P<0.05)；过表达 lncRNA GMDS-AS1组细胞具有与敲减 ESET组相似的上述检测指标的变化。 RIP实验显示， ESET与 lncRNA GMDS-AS1具有结合能力。结论甲基转移酶 ESET参与肝癌细胞的增殖、凋亡和糖酵解过程，其潜在的机制可能与负向调节 lncRNA GMDS-AS1有关。 |
| 英文摘要: |
| Objective To explore the regulation of histone methyltransferase SET domain bifurcated 1 (ESET) and long noncoding RNA GDP-mannose 4, 6-dehydratase antisense RNA1 (lncRNA GMDS-AS1) on hepatoma cell proliferation, apoptosis and glycolysis and the potential relationship between them.Methods A total of 49 cases of hepatocellular carcinoma tissues and adjacent tissues sur.gically resected from the Affiliated Hospital of Youjiang Medical University and Baise People's Hospital for Nationalities between Janu.ary 2019 and January 2020 were collected. The blank group (without any treatment), empty group (transfected with pcDNA or si-con), ESET knockdown group (transfected with si-ESET) and lncRNA GMDS-AS1 overexpression group (transfected with pcDNA-GMDS-AS1) were transfected into HepG2 cells by the liposome method. The expression levels of ESET, lncRNA GMDS-AS1, cell proliferation antigen marker Ki-67, glucose transporter 1 (GLUT-1), lactate dehydrogenase A (LDHA) and activated cysteine protease (C-caspase-3) were detected by real-time fluorescence quantitative reverse transcription-polymerase chain reaction (qRT.PCR) and Western blotting. Cell proliferation and apoptosis were detected by the cell counting kit (CCK-8) method and flow cytometry. An RNA immunocoprecipi.tation (RIP) assay was used to detect the relationship between ESET and lncRNA GMDS-AS1.Results The expression levels of ESETmRNA and protein in hepatocellular carcinoma tissues were 5.18±0.94 and 0.69±0.14, respectively, which were significantly higherthan those in adjacent tissues [(0.99±0.33) and (0.23±0.07)], and the expression level of lncRNA GMDS-AS1 in hepatocellular carcino. ma tissues was 0.63±0.20, which was significantly lower than that in adjacent tissue (1.02±0.31) (P < 0.05), and both were negatively correlated. Compared with the empty vector group, the expression levels of ESET, Ki-67, GLUT-1 and LDHA protein as well as cell ac. tivity in the knockdown ESET group were significantly decreased, and C-caspase-3 protein expression and apoptosis rate were signifi. cantly increased (P < 0.05). The lncRNA GMDS-AS1 overexpression group had similar index changes as the ESET knockdown group.RIP experiments showed that ESET had the ability to interact lncRNA GMDS-AS1.Conclusion The histone methyltransferase ESETwas involved in the proliferation, apoptosis, and glycolysis processes of hepatoma cells, and the underlying mechanism may be relatedto the negative regulation of lncRNA GMDS-AS1. |
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