ENO3在肝癌中的表达及其对奥沙利铂化疗敏感度的影响

崔红磊; 张笑丹; 郭丹风; 闫志平; 郭文治; 张水军

doi:10.3971/j.issn.1000-8578.2022.21.1020

ENO3在肝癌中的表达及其对奥沙利铂化疗敏感度的影响

Expression of ENO3 and Its Effect on Sensitivity of Hepatocellular Carcinoma Cells to Oxaliplatin

摘要

摘要:
目的研究ENO3基因在肝癌中的表达及其是否可影响肝癌细胞对奥沙利铂的敏感度并探讨可能机制。
方法 qRT-PCR、免疫组织化学法检测48例肝细胞肝癌组织和正常肝组织中ENO3的表达。构建ENO3过表达质粒, 转染MHCC97H细胞和HepG2细胞; 实验分为空载组(Vector组)、ENO3过表达组(ENO3组)、空载+OXA组(Vector+OXA组)和ENO3过表达+OXA组(ENO3+OXA组)。CCK-8法、克隆形成法检测细胞增殖能力, 流式细胞术检测细胞凋亡率; Western blot法检测细胞中凋亡相关蛋白Bcl-2、Bax和Caspase-3的表达水平。
结果肝癌组织中ENO3的表达量明显低于正常肝组织。ENO3过表达组细胞ENO3的mRNA水平与蛋白水平较空载组均明显升高。过表达ENO3联合奥沙利铂后, 与空载+OXA组相比, ENO3过表达+OXA组细胞活力降低、凋亡增加; Bcl-2蛋白表达减少, Bax、Caspase-3蛋白表达增加。
结论 ENO3在肝癌组织中表达下降, 过表达ENO3可通过促进凋亡增强肝癌细胞对奥沙利铂的敏感度。

Abstract:
Objective To investigate the expression of ENO3 gene in hepatocellular carcinoma and its effect on the sensitivity of hepatocellular carcinoma cell lines to OXA, and to explore the possible mechanism.
Methods qRT-PCR and immunohistochemical analysis were used to detect the expression of ENO3 in 48 pairs of hepatocellular carcinoma tissues and normal liver tissues.Overexpression plasmid was constructed and transfected into MHCC97H and HepG2 cells.The experiments were divided into empty group (Vector group), ENO3 overexpression group (ENO3 group), empty+OXA group (Vector+OXA group) and ENO3 overexpression+OXA group (ENO3+OXA group).The proliferation ability of MHCC97H and HepG2 cells were detected by CCK-8 assay and cell colony formation assay.The apoptosis rate was determined by flow cytometry assay.Protein expressions of Bcl-2, Bax and Caspase-3 were detected by Western blot assay.
Results The expression of ENO3 was significantly decreased in hepatocellular carcinoma tissues, compared with normal liver tissues adjacent to the carcinoma.The expression of ENO3 gene in the ENO3 overexpression group was significantly higher than that in the empty group.Compared with the Vector+OXA group, cell viability was decreased, apoptosis rate was increased, Bcl-2 protein expression was decreased, Bax and Caspase-3 protein expression were increased in the ENO3+OXA group.
Conclusion The expression of ENO3 is down-regulated in hepatocellular carcinoma tissues, and the overexpression of ENO3 can enhance the sensitivity of hepatocellular carcinoma cell lines to oxaliplatin by promoting cell apoptosis.

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