国际遗传学杂志
國際遺傳學雜誌
국제유전학잡지
INTERNATIONAL JOURNAL OF GENETICS
2012年
1期
1-5
,共5页
刘静蕾%姜季%赵红丽%李欣%浦海宏%王艳
劉靜蕾%薑季%趙紅麗%李訢%浦海宏%王豔
류정뢰%강계%조홍려%리흔%포해굉%왕염
人肺腺癌%Survivin%SiRNA%凋亡%裸鼠
人肺腺癌%Survivin%SiRNA%凋亡%裸鼠
인폐선암%Survivin%SiRNA%조망%라서
Human lung adenocarcinoma%Survivin%SiRNA%Apoptosis%Nude mice
目的 探讨靶向survivin基因特异性siRNA对人肺腺癌细胞株AGZY的凋亡、增殖影响以及对其相关机制的探讨.方法 采用RNAi技术靶向干扰AGZY细胞survivin基因的表达,分空白组、转染试剂组、阴性对照组、转染组,用RT-PCR检测各组survivin基因的表达,绘制细胞生长曲线,Hoechst染色检测各组细胞凋亡情况,Western印迹检测各组蛋白survivin、caspase-3、caspase-8、cyclinB1、cyclinD1的变化.结果 Survivin特异性siRNA可在mRNA及蛋白水平有效下调survivin的表达,转染组与其他3组比较差异有统计学意义(F=1785.25,F=38.67,P＜0.05)；survivin表达抑制后,与其他3组比较,转染组AGZY细胞数明显减少(F=14999.46,P＜0.05),且转染组细胞出现典型的凋亡改变；与其他3组相比较,survivin特异性siRNA转染组cyclinB1、cyclinD1表达下调(F=109.674,F=313.102,P＜0.05),caspase-3、caspase-8表达上调(F=101.60,F=782.504,P＜0.05).结论 Survivin基因特异性siRNA可以有效下调survivin的表达；survivin表达下降可促进细胞凋亡,抑制细胞增殖；细胞增殖受抑制可能与cyclinB1、cyclinD1表达下降有关,细胞凋亡增加可能与caspase-3、caspase-8表达上调有关.
目的 探討靶嚮survivin基因特異性siRNA對人肺腺癌細胞株AGZY的凋亡、增殖影響以及對其相關機製的探討.方法 採用RNAi技術靶嚮榦擾AGZY細胞survivin基因的錶達,分空白組、轉染試劑組、陰性對照組、轉染組,用RT-PCR檢測各組survivin基因的錶達,繪製細胞生長麯線,Hoechst染色檢測各組細胞凋亡情況,Western印跡檢測各組蛋白survivin、caspase-3、caspase-8、cyclinB1、cyclinD1的變化.結果 Survivin特異性siRNA可在mRNA及蛋白水平有效下調survivin的錶達,轉染組與其他3組比較差異有統計學意義(F=1785.25,F=38.67,P＜0.05)；survivin錶達抑製後,與其他3組比較,轉染組AGZY細胞數明顯減少(F=14999.46,P＜0.05),且轉染組細胞齣現典型的凋亡改變；與其他3組相比較,survivin特異性siRNA轉染組cyclinB1、cyclinD1錶達下調(F=109.674,F=313.102,P＜0.05),caspase-3、caspase-8錶達上調(F=101.60,F=782.504,P＜0.05).結論 Survivin基因特異性siRNA可以有效下調survivin的錶達；survivin錶達下降可促進細胞凋亡,抑製細胞增殖；細胞增殖受抑製可能與cyclinB1、cyclinD1錶達下降有關,細胞凋亡增加可能與caspase-3、caspase-8錶達上調有關.
목적 탐토파향survivin기인특이성siRNA대인폐선암세포주AGZY적조망、증식영향이급대기상관궤제적탐토.방법 채용RNAi기술파향간우AGZY세포survivin기인적표체,분공백조、전염시제조、음성대조조、전염조,용RT-PCR검측각조survivin기인적표체,회제세포생장곡선,Hoechst염색검측각조세포조망정황,Western인적검측각조단백survivin、caspase-3、caspase-8、cyclinB1、cyclinD1적변화.결과 Survivin특이성siRNA가재mRNA급단백수평유효하조survivin적표체,전염조여기타3조비교차이유통계학의의(F=1785.25,F=38.67,P＜0.05)；survivin표체억제후,여기타3조비교,전염조AGZY세포수명현감소(F=14999.46,P＜0.05),차전염조세포출현전형적조망개변；여기타3조상비교,survivin특이성siRNA전염조cyclinB1、cyclinD1표체하조(F=109.674,F=313.102,P＜0.05),caspase-3、caspase-8표체상조(F=101.60,F=782.504,P＜0.05).결론 Survivin기인특이성siRNA가이유효하조survivin적표체；survivin표체하강가촉진세포조망,억제세포증식；세포증식수억제가능여cyclinB1、cyclinD1표체하강유관,세포조망증가가능여caspase-3、caspase-8표체상조유관.
Objective To investigate the effects of survivin-specific siRNA on the proliferation and apoptosis of human lung adenocarcinoma cell line AGZY,and to explore their mechanisms.Methods Survivin-specific siRNA was tranfected into AGZY cells.Four groups were included:negative control group,DharmaFECT-treated group,nonsense siRNA-treated group,survivin specific siRNA-treated group.The expression of survivin mRNA levels was determined using RT-PCR.Cell growth was studied.The apoptosis was analyzed by Hoechst 33258 staining method.The protein expression levels was determined using Western blotting analyses.Results Campared with other three groups,survivin-specific siRNA could efficiently down-regulate the survivin expression at mRNA and protein levels(F =1785.25,F =38.67,P ＜ 0.05).After transfection,the cell proliferation decreased significantly compared with other three groups(F =14999.46,P ＜ 0.05).The typical changes associated with apoptosis was observed in the transfected group.Compared with other three groups,in transfected group,the expression of caspase-3 and caspase-8 was upre gulated(F =101.60,F =782.504,P ＜ 0.05),and the expression of cyclinB1 and cyclinD1 was downregulated(F =109.674,F =313.102,P ＜ 0.05).Conclusions Survivin-specific siRNA can supresses the expression of survivin,increase apoptosis,and inhibit cells proliferation.It may have relationship with the upregulation of caspase-3 and caspase-8,and the downregulation of cyclinB1 and cyclinD 1.
