[1]汪欣,杨军,许波,等.survivin基因沉默对腺样囊性癌细胞生长和凋亡的影响[J].第三军医大学学报,2009,31(11):1053-1056.
 WANG Xin,YANG Jun,XU Bo,et al.Effects of survivin gene silencing on growth and apoptosis of adenoid cystic carcinoma cell line[J].J Third Mil Med Univ,2009,31(11):1053-1056.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第11期
页码:
1053-1056
栏目:
论著
出版日期:
2009-06-15

文章信息/Info

Title:
Effects of survivin gene silencing on growth and apoptosis of adenoid cystic carcinoma cell line
作者:
汪欣杨军许波熊宇张从纪
第三军医大学西南医院口腔科
Author(s):
WANG Xin YANG Jun XU Bo XIONG Yu ZHANG Cong-ji
Department of Stomatology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China
关键词:
RNA干扰腺样囊性癌细胞株survivincaspase-3
Keywords:
RNA interference adenoid cystic carcinoma cells line survivin caspase-3
分类号:
R394.2;R73-362;R730.23
文献标志码:
A
摘要:
目的   探讨RNA干扰下调survivin基因表达后腺样囊性癌细胞生物学行为变化及其与caspase-3基因表达的相关性。   方法   设计、合成小干扰RNA(small interfering RNA,siRNA),构建表达载体pGenesil-shRNA-survivin,将重组质粒导入人腺样囊性癌ACC-2细胞株,G418筛选阳性克隆 。采用逆转录-聚合酶链反应(RT-PCR)、Western blot分别检测转染后的ACC-2细胞中survivin、caspase-3 mRNA及蛋白表达的变化,用噻唑蓝(MTT)法检测细胞的增殖,原位细胞凋亡(TUNEL)法及透射电子显微镜研究细胞凋亡。   结果   测序证实表达载体构建成功,在pGenesil-shRNA-survivin的ACC-2细胞株中 , survivin mRNA及蛋白表达明显降低(P<0.05), caspase-3 mRNA及蛋白表达明显增加(P<0.05),survivin与caspase-3的mRNA和蛋白表达呈显著负相关(r=-0.333, P<0.05)。ACC-2细胞增殖受到抑制,透射电镜及TUNEL法可见细胞出现凋亡。   结论   成功构建了survivin基因的RNAi真核表达载体,pGenesil-shRNA-survivin重组质粒能有效抑制腺样囊性癌细胞生长。
Abstract:
Objective   To investigate the effects of survivin down-regulation on the biological behavior of adenoid cystic carcinoma cells and the relationship with caspase-3.     Methods   Small interfering RNA (siRNA) was designed and synthesized for the construction of eukaryotic expression vector pGenesil-shRNA-survivin. The recombinant plasmid was transfected into ACC-2 cells. The positive clones were screened with G418. The expressions of survivin and caspase-3 mRNA and protein were detected by RT-PCR and Western blot. The proliferation of adenoid cystic carcinoma cells was detected by MTT. Cell apoptosis was detected by TUNEL assay and transmission electron microscopy.     Results   The recombinant plasmid was successfully constructed. Down-regulation was found in the expressions of survivin mRNA and protein (P<0.05), but up-regulation in the expressions of caspase-3 mRNA and protein (P<0.05). There was a negative correlation between the mRNA and protein expression of caspase-3 and survivin (r=-0.333, P<0.05). ACC-2 cell proliferation was inhibited. TUNEL assay and transmission electron microscopy showed typical apoptosis of cells.     Conclusion    The RNAi eukaryotic vector pGenesil-shRNA-survivin was constructed successfully. Recombinant plasmid pGenesil-shRNA-survivin can inhibit the proliferation of adenoid cystic carcinoma cells.

参考文献/References:

汪欣, 杨军, 许波, 等. survivin基因沉默对腺样囊性癌细胞生长和凋亡的影响[J]. 第三军医大学学报,2009,31(11):1053-1056.

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更新日期/Last Update: 2009-05-21