文献类型：期刊文献

中文题名：组蛋白去乙酰化酶抑制剂DWP0016对肺癌A549细胞和小鼠Lewis肺癌的抑制作用

英文题名：Antitumor activity of histone deacetylases inhibitor,DWP0016,in lung carcinoma A549 cells and xenograft mouse model

作者：金惠[1,2];刘力锋[3];邓卫平[3];刘建文[1,3]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237;[2]同济大学生命科学与技术学院,上海200092;[3]华东理工大学药学院,上海200237

年份：2013

卷号：27

期号：4

起止页码：657

中文期刊名：中国药理学与毒理学杂志

外文期刊名：Chinese Journal of Pharmacology and Toxicology

收录：CSTPCD;;Scopus;北大核心:【北大核心2011】；CSCD:【CSCD2013_2014】；

基金：上海市科学技术委员会基础研究重点项目(09JC1404500)~~

语种：中文

中文关键词：组蛋白去乙酰化酶抑制剂;肺癌;10号染色体同源丢失性磷酸酶张力蛋白;磷酸化Akt

外文关键词：histone deacetylases inhibitor; lung cancer; phosphatase and tensin homolog deleted on chromosome 10; phosphorylated-Akt

摘要：目的探讨新型组蛋白去乙酰化酶(HDAC)抑制剂DWP0016对肺癌的抑制作用及可能机制。方法①体外实验:DWP0016 0.625～10μmo.lL-1作用A549细胞48 h,MTT法检测细胞存活,实时荧光定量PCR检测10号染色体同源丢失性磷酸酶张力蛋白(PTEN)mRNA水平;Western蛋白质印迹法测定总组蛋白H3,乙酰化H3(Ac-H3),细胞周期调控因子p21,PTEN,总Akt和磷酸化Akt(p-Akt)的表达。②在体实验:小鼠腋下接种0.2 ml Lewis肿瘤细胞悬液制备肺癌模型,7 d后按照分组分别ip给予DWP001612.5,25和50 mg.kg-1及伏林司他(SAHA)50 mg.kg-1,每天1次,连续8 d。取肿瘤组织称重,计算肿瘤抑制率;免疫组化法检测肿瘤组织中Ac-H3和PTEN的表达。结果①体外实验:MTT结果显示,DWP0016抑制A549细胞增殖的IC50为2.51μmol.L-1,SAHA IC50为6.03μmol.L-1。与正常对照组相比,DWP0016 2.5μmo.l L-1组细胞组蛋白H3乙酰化及p21蛋白表达显著上升(P<0.05),PTEN的转录水平和蛋白水平上调,Akt的磷酸化水平下调(P<0.05)。②在体实验:与模型组比较,给予DWP001612.5 mg.kg-1移植瘤小鼠的瘤质量显著降低(P<0.05),抑瘤率达42.0%,给予DWP0016 50 mg.kg-1肿瘤抑制率高于SAHA 50 mg.kg-1(P<0.05);与模型组比较,肿瘤组织中Ac-H3和PTEN的表达增加。结论 DWP0016能明显抑制A549细胞增殖和Lewis肺癌模型小鼠的肿瘤生长,其机制与诱导Ac-H3的表达、激活p21、促进抑癌因子PTEN的转录及下调Akt的磷酸化水平有关。
OBJECTIVE To study the effect and the underlying molecular mechanisms of a novel histone deacetylases（HDAC） inhibitor named DWP0016 in lung carcinoma A549 cells and xenografts.METHODS ① In vitro： A549 cells were treated with DWP0016 0.625-10 μmol·L-1 for 48 h before cell viability was detected by MTT assay.The effect of DWP0016 on mRNA level of phosphatase and tensin homolog deleted on chromosome 10（PTEN） was detected by real-time PCR.The protein expression of total H3,acetylated-H3（Ac-H3）,p21,PTEN,total-Akt and phosphorylated-Akt（p-Akt） was detected by Western blotting.② In vivo： Lewis lung cancer animal models were constructed by injecting the axillae with 0.2 ml tumor suspension.After 7 d,the xenografts groups were ip given DWP0016 12.5,25 and 50 mg·kg-1 and positive drug vorinostat（SAHA） 50 mg·kg-1,once daily,for 8 d.After treatment,the animals were sacrificed to get the tumors.The tumor specimens were weighed and the tumor growth inhibition was calculated.The expression of Ac-H3 and PTEN in the specimen was detected by immunohistochemical assay.RESULTS ① In vitro： MTT results showed that IC50 of DWP0016 was 2.51 μmol·L-1 and IC50 of SAHA was 6.03 μmol·L-1.Compared with normal control,DWP0016 significantly up-regulated the protein expression of Ac-H3 and p21 at the concentration of 2.5 μmol·L-1（P 0.05）.DWP0016 promoted the mRNA level and protein expression of PTEN while decreasing the expression of p-Akt concentration-dependently（P0.05）.② In vivo： compared with model group,the mean mass of the specimens from DWP0016 12.5 mg·kg-1 significantly decreased（P0.05） with a 42.0% tumor growth inhibition.The tumor growth inhibition in DWP0016 50 mg·kg-1 group was higher than in SAHA 50 mg·kg-1 group（P0.05）.Compared with the model group,the expression of Ac-H3 and PTEN in tumor specimens was up-regulated by DWP0016.CONCLUSION DWP0016 effectively inhibits the proliferation in A549 cells and the tumor growth in xenografts.The molecular mechanisms are associated with the induction of Ac-H3,the activation of p21,PTEN and down-regulation of p-Akt.DWP0016 is a potent compound to be developed as an anti-tumor agent for clinic application in the future.