1270 《癌症》 Chinese Journal of Cancer,2009,28(12):1270-12 基础研究 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 张东1,刘长庭1,于晓妉2,刘岩 Synergistic cytotoxicity effect of histone deacetylase inhibitor combined with paclitaxel on lung cancer cell lines and its mechanism Dong Zhang, ' Chang-Ting Liu, Xiao-Dan Yu? and Yan Liu' [ Abstract] Background and Objective: Histone deacetylase (HDAC inhibitors 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 1.解放军总医院南楼 and to investigate its mechanism. Methods: H322 and H1299 cells were 呼吸科 divided into control group, paclitaxel (TAX) group, TSa group, and 北京100853 combination group(TF group, TSa followed by paclitaxel ). Cell proliferation 2.解放军军事医学科学院 was determined by MTT assay. Cell cycle and apoptosis were determined by 基础所 flow cytometry. The protein expression levels of survivin, ERK, and PARP 北京100850 were determined by Western blot analysis. Results: When combined with TSA, the 50% inhibition concentration (ICso) of paclitaxel decreased from 1. Geriatric Respiratory (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 P R. China TF group of H1299 cell line than in the other groups. pERK was up-regulated 2. Institute of Basic Medical 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 down- Academy of Military Medical regulated in the TSA and tF groups of two cell lines. Cleaved PARP was Science, Beijing 100850 detected in the TAX and the tF groups of H322 cells, and its expression P R. China 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 通讯作者:张东 combined with paclitaxel has a synergistic cytotoxicity effect on lung cancer Correspondence to: Dong Zhang cell lines H322 and H1299 when the cells were treated with TSA followed by Tel.:86.013521291696 paclitaxel. The mechanism may be that TSA down-regulates the survivin high- gd1117@yahoo.com expression induced by paclitaxel, and blocks pERK protein expression Key words: lung cancer, histone deacetylase, trichostatin A, paclitaxel 基金项目:军队“十一五”面上课 apoptosis, ERK 题B类 【摘要】背景与目的:组蛋白去乙酰化酶( histone deacetylase,HDAC)抑制剂 Grant: Army "1lth Five-Year 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 Plan"Surface Project Class B 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 收稿日期:2009-06-10 H322及H299的作用及机制。方法:将H322和H1299细胞分别分成4组:(1) 修回日期:2009-08-07 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!癌症" !"#$%&% ’()*$+, (- .+$/%*# 0112# 03$40%&506784069 !基础研究! !!"#$%&’$" (&’)*%+,-. &-. /"01’$231# !"#$%&’ ()*+)$,-*#) $./01% "&2"3"$%4# +5& "&2"6"$ +)-- #"7&5- &)$8%49 :;&<$"%& $24%;72 ()<4)5#"&7 )=>4’##"%& %: ?;-$">-’ 7’&’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‘JaOKJ% &?%- b c $% $aOI] ‘XO_J% &?%- b c "& .IJJ +)--# 5&( :4%? $KKYOa‘I^O_% &?%- b c $% $aIO_‘KKO^% &?%- b c "& .KJLL +)--#& 8"$2 #"7&":"+5&$ ("::)4)&+)# $!dYOYX%O 0>%>$%#"# 45$) %: .IJJ +)--# 85# 2"72)4 "& $2) $2) EN 74%;> $25& "& $2) E0M 74%;>$!dYOYX%O E2)4) 8)4) ?%4) &)+4%#"# +)--# "& $2) EN 74%;> %: .KJLL +)-- -"&) $25& "& $2) %$2)4 74%;>#O >QeS 85# ;>F4)7;-5$)( "& $2) E0M 74%;> %: .IJJ +)-- -"&)O Q=>4)##"%& %: H;4A"A"& 85# ;>F4)7;-5$)( "& $2) E0M 74%;> %: $8% +)--#O Q=>4)##"%&# %: H;4A"A"& 5&( >QeS 8)4) (%8&F 4)7;-5$)( "& $2) EH0 5&( EN 74%;># %: $8% +)-- -"&)#O D-)5A)( fGef 85# ()$)+$)( "& $2) EGM 5&( $2) EN 74%;># %: .IJJ +)--#& 5&( "$# )=>4)##"%& 85# #"7&":"+5&$-, 2"72)4 "& $2) $2) EN 74%;> $25& "& $2) EGM 74%;>O D-)5A)( fGef 85# &%$ ()$)+$)( "& )5+2 74%;> %: .KJLL +)--#O 8+-’7,#2+-## EHG +%?6"&)( 8"$2 >5+-"$5=)- 25# 5 #,&)47"#$"+ +,$%$%="+"$, )::)+$ %& -;&7 +5&+)4 +)-- -"&)# .IJJ 5&( .KJLL 82)& $2) +)--# 8)4) $4)5$)( 8"$2 EHG :%--%8)( 6, >5+-"$5=)-O E2) ?)+25&"#? ?5, 6) $25$ EHG (%8&F4)7;-5$)# $2) #;4A"A"& 2"72F )=>4)##"%& "&(;+)( 6, >5+-"$5=)-& 5&( 6-%+9# >QeS >4%$)"& )=>4)##"%&O 91: ;+%.## -;&7 +5&+)4& 2"#$%&) ()5+)$,-5#)& $4"+2%#$5$"& G& >5+-"$5=)-& 5>%>$%#"#& QeS !摘 要" 背景与目的#组蛋白去乙酰化酶$"#&:($% ;%+/%:<,+&%& =>?.%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’ =>?. 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 =>?. 抑制剂曲古抑菌素 ?$:*#/"(&:+:#$ ?& @A?%协同紫杉醇抑制肺癌细胞 =B00 及 =5022 的作用及机制’ 方法#将 =B00 和 =5022 细胞分别分成 C 组#$5% 对照组($0%紫杉醇组$@?D%($B%@A? 组($C%以 @A? 预先作用 50 " 后&再使用紫 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 张 东 5 " 刘长庭 5 " 于晓妉 0 " 刘 岩 5 <:-1%*2#$2’ ’:$+$+=2’2$: 1>>1’$ +> 52#$+-1 .1&’1$:7&#1 2-52"2$+% ’+?"2-1. ;2$5 @&’72$&=17 +- 7,-* ’&-’1% ’177 72-1# &-. 2$# ?1’5&-2#? /%&7 g2*&7&K D2*&7FE"&7 c";&K M"*%F/*& h;J *&( h*& c";K 5E 解放军总医院南楼 呼吸科" 北京 4773FB 0G 解放军军事医学科学院 基础所" 北京 4773F7 !" #$%&’(%&) *$+,&%’(-%. /$,’%(0$1(" 234 #$1$%’5 6-+,&(’5" 7$&8&19 !::;<=" 2" *" >?&1’ @" A1+(&(B($ -C 7’+&) D$E&)’5 F)&$1)$+" 4)’E$0. -C D&5&(’%. D$E&)’5 F)&$1)$" 7$&8&19 !::;<:" 2" *" >?&1’ 通讯作者# 张 东 .(**%&H($;%$/% :(# I($J K"+$J @%,G# 39G74BF04024929 LM+#,# N"+$J;5556O<+"((G/(M 基 金 项 目 #军 队 $十 一 五 %面 上 课 题 P 类 !"#$%# ?*M< $55:" Q#R%ST%+* U,+$% A)*-+/% U*(V%/: .,+&& P 收稿日期#0772S79S57 修回日期#0772S73S76 !"#$ C M Y K