组蛋白去乙酰化酶抑制剂协同紫杉醇对人肺癌细胞株抑制作用及机制  

Synergistic cytotoxicity effect of histone deacetylase inhibitor combined with paclitaxel on lung cancer cell lines and its mechanism

作　　者：张东[1] 刘长庭[1] 于晓妉[2] 刘岩[1] 

机构地区：[1]解放军总医院南楼呼吸科,北京100853 [2]解放军军事医学科学院基础所,北京100850

出　　处：《癌症》2009年第12期1270-1276,共7页Chinese Journal of Cancer

基　　金：军队"十一五"面上课题B类~~

摘　　要：背景与目的:组蛋白去乙酰化酶(histone deacetylase,HDAC)抑制剂通过抑制多种基因或蛋白介导的信号转导网络的功能,影响细胞增殖及对化疗药物的敏感性。HDAC抑制剂可能会提高紫杉醇对肺癌细胞的抑制作用。本研究评价HDAC抑制剂曲古抑菌素A(trichostatin A,TSA)协同紫杉醇抑制肺癌细胞H322及H1299的作用及机制。方法:将H322和H1299细胞分别分成4组:(1)对照组;(2)紫杉醇组(TAX);(3)TSA组;(4)以TSA预先作用12h后,再使用紫杉醇的联合用药组(TF)。分别以MTT法、流式细胞术检测细胞增殖情况以及细胞周期、细胞凋亡等指标的变化,荧光显微镜观察细胞核形态的改变,Western blot检测Survivin、PARP蛋白及细胞外信号调节激酶(ERK)的表达。结果:TSA明显增强了紫杉醇对两种肺癌细胞的抑制率。紫杉醇作用96h对H322细胞的IC50由(48.07±26.12)nmol/L下降至(6.34±5.72)nmol/L,对H1299细胞的IC50由(110.6±38.7)nmol/L下降至(63.7±11.8)nmol/L,差异具有统计学意义(P<0.05)。TF组H322细胞凋亡率较TAX组明显增加(P<0.05);各组H1299细胞出现少量凋亡细胞,TF组的死亡细胞明显多于其他组。TAX上调H322细胞中pERK的表达,TSA及TF减低两种细胞中pERK表达;TAX明显增加Survivin的表达,TSA及TF下调了Survivin的表达。TAX组和TF组H322细胞中均检测到剪切PARP蛋白,TF组较TAX组剪切PARP蛋白明显增加,而H1299细胞各组均未检测到剪切PARP蛋白。结论:HDAC抑制剂TSA联合紫杉醇可以协同抑制H322及H1299细胞增殖,促进H322细胞凋亡和H1299细胞死亡。协同机制可能与通过下调肺癌细胞由紫杉醇诱发的Survivin高表达和阻断ERK通路的激活有关。Background and Objective： Histone deacetylase （HDAC） inhibiters can inhibit cell signal network function through decreasing expression of multiple genes and proteins, thus affect cell proliferation, survival and chemosensitivity. HDAC inhibitors combined with paclitaxel may enhance the inhibitory effect of drugs on lung cancer cells. This study was to observe the synergistic anti-proliferative effect of HDAC inhibitor trichostatin A （TSA） combined with paclitaxel on lung cancer cell lines H322 and H1299, and to investigate its mechanism. Methods： H322 and H1299 cells were divided into control group, paclitaxel （TAX） group, TSA group, and combination group （TF group, TSA followed by paclitaxel）. Cell proliferation was determined by MTT assay. Cell cycle and apoptosis were determined by flow cytometry. The protein expression levels of survivin, ERK, and PARP were determined by Western blot analysis. Results. When combined with TSA, the 50% inhibition concentration （IC50） of paclitaxel decreased from （48.07±26.12） nmol/L to （6.34±5.72） nmol/L in H322 cells and from （110.6±38.7） nmol/L to （63.7±11.8） nmol/L in H1299 cells, with significant differences （P〈0.05）. Apoptosis rate of H322 cells was higher in the the TF group than in the TAX group（P〈0.05）. There were more necrosis cells in the TF group of H1299 cell line than in the other groups, pERK was up-regulated in the TAX group of H322 cell line. Expression of Survivin was up-regulated in the TAX group of two cells. Expressions of Survivin and pERK were downregulated in the TSA and TF groups of two cell lines. Cleaved PARP was detected in the TAX and the TF groups of H322 cells, and its expression was significantly higher in the the TF group than in the TAX group. Cleaved PARP was not detected in each group of H1299 cells. Conclusions. TSA combined with paclitaxel has a synergistic cytotoxicity effect on lung cancer cell lines 1-1322 and 141299 when the cells were treated with TSA followed by paclitaxel. The me

关 键 词：肺癌 组蛋白去乙酰化酶 曲古抑菌素A 紫杉醇 细胞凋亡 细胞外信号通路激酶 

分 类 号：R734.2[医药卫生—肿瘤]