组蛋白去乙酰化酶2表达下调对喉鳞癌Hep-2细胞凋亡和增殖及迁移的影响

Down-regulation of histone deacetylase 2 induces cell apoptosis and inhibits cell proliferation and migration of laryngeal squamous cell carcinoma cells

目的探讨组蛋白去乙酰化酶2（HDAC2）表达下调对喉鳞癌Hep-2细胞增殖、细胞凋亡和细胞迁移的影响。方法利用脂质体2000将HDAC2小干扰RNA（siRNA）和对照siRNA转染喉鳞癌Hep-2细胞，将细胞分为3组：未处理组、siRNA对照组和HDAC2siRNA转染组。利用Westernblot检测转染HDAC2siRNA后喉鳞癌Hep-2细胞中HDAC2蛋白的表达；利用CCK-8分析3组细胞增殖的变化；采用流式细胞术检测3组细胞凋亡情况；接着利用Boyden小室研究3组细胞迁移能力的变化，最后采用Westernblot研究与细胞凋亡和细胞迁移密切相关蛋白的表达。结果HDAC2siRNA明显下调喉鳞癌Hep-2细胞中HDAC2蛋白的表达。HDAC2表达下调明显抑制喉鳞癌Hep-2细胞的增殖和诱导细胞凋亡，并抑制喉鳞癌Hep-2细胞的迁移。此外，HDAC2表达下调明显增加caspase-3和caspase-9的表达，但降低基质金属蛋白酶（MMP）-2和MMP-9的表达。结论HDAC2可能在喉鳞癌发生发展中发挥重要作用，其表达下调介导的细胞凋亡和迁移抑制可能与caspase-3和caspase-9表达的升高以及MMP-2和MMP-9表达的降低相关。

Objective To investigate the effects of histone deacetylase 2 （HDAC2） expression on cell proliferation, apoptosis and migration of laryngeal squamous cell carcinoma （LSCC） Hep-2 cells. Methods HDAC2 siRNA and control siRNA were transfected into LSCC Hep-2 cells by lipofectamine 2000, and ceils were divided into three experimental groups： untreated group, control siRNA group and HDAC2 siRNA transfecfion group. Western blotting was utilized to detect the expression of HDAC2 protein in Hep-2 cells. Cell proliferation and apoptosis were investigated by CCK-8 kit and flow cytometry, respectively. Boyden chamber was used to study cell migration. Expressions of cell apoptosis and cell migration related proteins were detected by Western blotting. Results HDAC2 siRNA significantly down- regulated the expression of HDAC2 protein in LSCC Hep-2 cells. Down-regulation of HDAC2 expression coincided with an inhibition of cell proliferation and migration along with an induced cell apoptosis of Hep-2 cells. Moreover, down-regulation of HDAC2 expression significantly increased the expressions of caspase-3 and caspase-9 proteins but decreased the expressions of matrix metalloproteinases （ MMP）-2 and MMP-9 proteins. Conclusions HDAC2 may play a pivotal role in the initiation and development of LSCC. Downregulation of HDAC2 expression mediates cell apoptosis. Cell migration inhibition may be tightly associated with overexpression of caspase-3 and caspase-9 along with down-regulation of MMP-2 and MMP-9 expressions.