HDAC1 siRNA转染对肺腺癌NCI-H1299细胞增殖及凋亡的影响

Role of HDAC1 RNAi in Proliferation and Apoptosis of Human Lung Adenocarcinoma Cell Line NCI-H1299

目的 通过HDACl siRNA转染肺腺癌细胞株NCI-H1299,观察HDACs中HDACl对肺腺癌细胞增殖、周期和凋亡的影响,明确HDACl在肺腺癌细胞生长中的作用.方法 MTT法检测不同时间点HDACl siRNA转染对NCI-H1299细胞体外增殖的影响,流式细胞术榆测RNA干扰后细胞周期及凋亡率的变化;Western印迹法检测细胞内组蛋白H4乙酰化水平的变化.结果 HDACl siRNA转染NCI-H1299细胞后,HDACl在转录和表达水平均下降,组蛋白H4乙酰化表达增高.siRNA干扰后NCI-H1299细胞的体外生长抑制呈时间依赖性,流式细胞术检测结果显示细胞阻滞于G2/M期,细胞凋亡增加.结论 HDACl siRNA转染能有效地抑制HDACl在NCI-H1299细胞中的表达,增加NCI-H1299细胞中组蛋白的乙酰化,并影响NCI-H1299细胞相关生物学行为,抑制肺腺癌细胞的生长,为HDACl基因治疗应用于肺腺癌奠定了基础.

Objective The role of HDACI in tumor cell proliferation was investigated using RNA interference-mediated protein knockdown,to determine whether inhibition of histone deacetylasel leads to tumor cell growth inhibition or leads to tumor cell death in vitro. Methods MTT assay was employed to evaluate the inhibitory effect of HDAC1 RNAi on growth of human lung adenocarcinoma cell line NCI-H1299 in vitro. The cell cycle distribution and apoptotic ratio were determined by flow cytometry.The acetyl level of histone H4 was detected by western blot. Results HDACI RNAi inhibited the growth of NCI-H1299 ceils in a timedependent manner.Flow cytometry showed that the cells were blocked at G2/M phase and cell apoptosis was increased compared to the control.By HDAC1 RNAi the acetyl level of histone H4 significantly increased in NCI-Hl299 cells. Conclusions The inhibitory effect on HDAC1 expression of siRNA on NCI-H1299 cells was confirmed,which inhibited tumour cell growth.This may serve as a biological basis for the application in the gene therapy of lung neoplasms.