SETD8抑制剂UNC0379对肝癌细胞增殖、迁移的影响

Effect of SETD8 inhibitor UNC0379 on the proliferation and migration of hepatic carcinoma cells

目的:探究组蛋白修饰酶SETD8在人肝癌细胞系Huh7中的表达,以及SETD8抑制剂UNC0379对肿瘤细胞增殖、迁移过程的影响。方法:RT-PCR和蛋白免疫印迹法检测人正常肝细胞系LO2和肝癌细胞系Huh7中SETD8的mRNA和蛋白水平,并分析GEO和TCGA数据库中SETD8在肝癌组织的表达。用不同浓度SETD8抑制剂UNC0379处理Huh7细胞后检测SETD8和H4K20me1蛋白表达水平,并通过细胞克隆、CCK-8、划痕和Transwell实验检测细胞的增殖和迁移能力。结果:与LO2相比,Huh7细胞中SETD8的mRNA和蛋白水平均增加。通过对GEO数据库GSE87630和TCGA数据库的分析,发现SETD8在HCC患者肝癌组织中的表达均显著增加。SETD8抑制剂处理后,Huh7细胞中SETD8和H4K20me1的蛋白表达水平下降。CCK-8、细胞克隆结果显示UNC0379可抑制Huh7细胞的增殖过程;Transwell和划痕实验证明UNC0379可抑制Huh7细胞的迁移过程。结论:UNC0379可下调SETD8和H4K20me1表达,并抑制Huh7细胞的增殖、迁移,这种抑制可能与H4K20me1水平下降有关。

Objective:To explore the expression of histone modifying enzyme SETD8 in human hepatic carcinoma cell line Huh7 and the effect of SETD8 inhibitor UNC0379 on the proliferation and migration of tumor cells.Methods:RT-PCR and Western blot were used to examine the expression of SETD8in human normal liver cell line LO2 and hepatic carcinoma cell line Huh7.The GEO and TCGA database were also used to analysis the expression of SETD8 in hepatocellular carcinoma(HCC)patients.Western Blot was performed to measure the expression of SETD8 and H4K20me1 in Huh7 cells treated with SETD8 inhibitor UNC0379.The cell proliferation and migration of Huh7 cells were detected by colony formation,CCK-8,wound healing and Transwell assays.Results:Compared with in LO2cells,the mRNA and protein levels of SETD8 increased in Huh7 cells;while the expression of SETD8 in liver tissue of HCC patients was significantly increased in GEO and TCGA database.The protein levels of SETD8 and H4K20me1 decreased in Huh7 cells when treated with SETD8 inhibitor.CCK-8 and colony formation results showed that SETD8 inhibitor UNC0379 could inhibit the proliferation of Huh7 cells,while wound healing and Transwell assays suggested that UNC0379 could inhibit the migration of Huh7 cells.Conclusion:UNC0379 could decrease the expression of SETD8and H4K20me1,and inhibit the proliferation and migration of Huh7 cells,which may be related to the decrease of H4K20me1 level.