组蛋白去甲基化酶JMJD2E抑制肝癌细胞恶性进展的研究

Histone Demethylase JMJD2E Suppresses Malignant Progression of Liver Cancer Cells

目的:探讨JMJD2E在肝癌细胞中的作用机制,以及其与miR372之间的调控关系。方法:本研究中,选择了Hep3B人肝癌细胞并构建了JMJD2E过表达的细胞系。使用了多种实验技术,包括RT-PCR、Northern印迹法、Western blotting等,来检测JMJD2E及相关基因的表达,同时通过miRNA检测工具详细分析和验证了miR372的表达。此外,还进行了免疫沉淀实验、RNA免疫沉淀、染色质免疫沉淀和超级凝胶移位实验,以深入研究JMJD2E蛋白与核酸的相互作用和染色质水平的作用机制。最后,通过CCK8测定法和细胞菌落形成效率测定评估了肝癌细胞的增殖和生长能力,并在小鼠体内进行异种移植实验验证了研究结果。结果:JMJD2E过表达明显抑制了肝癌细胞的生长和增殖能力,包括细胞数量减少、集落形成效率下降以及细胞增殖率的减少。异种移植实验证实了这种抑制效应。此外,JMJD2E在表观遗传学上减弱miR372的表达。结论:本研究揭示了JMJD2E在肝癌细胞中的抑制作用,可能通过调控miR372的表达发挥关键作用。JMJD2E的过表达降低了miR372的表达水平,从而影响了肝癌细胞的生长和增殖。为肝癌治疗提供了新的潜在靶点和策略,有望改善肝癌的治疗效果。

Objective:To explores the mechanistic role of JMJD2E in liver cancer cells and its regulatory relationship with miR372.Methods:In this study,selected Hep3B human liver cancer cells and established a cell line overexpressing JMJD2E.Employed various experimental techniques including RT-PCR,Northern blotting,and Western blotting to assess the expression of JMJD2E and related genes.Additionally,conducted immunoprecipitation experiments,RNA immunoprecipitation,chromatin immunoprecipitation,and super gel shift assays to delve into the interactions between JMJD2E protein and nucleic acids,as well as its chromatin-level mechanisms.Finally,evaluated the proliferation and growth capabilities of liver cancer cells using CCK8 assay and colony formation efficiency assay,corroborating the findings through xenograft experiments in mice.Results:Overexpression of JMJD2E significantly inhibited the growth and proliferation capacity of liver cancer cells,including a decrease in cell count,reduced colony formation efficiency,and diminished cell proliferation rate.Xenograft experiments validated this inhibitory effect.Furthermore,JMJD2E epigenetically attenuated the expression of miR372.Conclusion:This study unveils the inhibitory role of JMJD2E in liver cancer cells,potentially acting through the regulation of miR372 expression.Overexpression of JMJD2E reduced the expression level of miR372,thereby affecting the growth and proliferation of liver cancer cells.This provides new potential targets and strategies for liver cancer treatment,with the potential to improve therapeutic outcomes.