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Ad-delE1b55kD-shRNA/Survivin-EGFP的构建及鉴定

, PP. 1293-1297

Keywords: 肿瘤增殖型腺病毒,RNA干扰,存活素,结肠癌

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Abstract:

目的构建特异性强,转染率高,针对人Survivin进行RNA干扰的肿瘤增殖型腺病毒载体(Ad-delE1b55kD-shRNA/Survivin-EGFP)。方法将针对人Survivin的shRNA的基因序列及EGFP基因序列克隆至肿瘤增殖型腺病毒穿梭质粒pAd-delE1b55kD;HEK-293细胞中包装出毒得到重组腺病毒Ad-delE1b55kD-shRNA/Survivin-EGFP并测序鉴定。用重组腺病毒感染结肠癌细胞HT-29并检测转染率,用RT-PCR和Westernblot分别检测转染后的HT-29细胞中SurvivinmRNA和蛋白表达的变化。结果序列测定证明针对人Survivin的shRNA序列及EGFP序列被成功构建到肿瘤增殖型腺病毒载体中;Ad-delE1b55kD-shRNA/Survivin-EGFP对结肠癌细胞HT-29的转染效率显著高于增殖缺陷型腺病毒载体和脂质体载体(P<0.01);而对肝细胞的转染率则显著低于对结肠癌细胞株HT-29的转染(P<0.01)。与复制缺陷型腺病毒载体和脂质体载体比较,转染Ad-delE1b55kD-shRNA/Survivin-EGFP后,HT-29细胞中SurvivinmRNA拷贝数及蛋白表达显著降低(P<0.01)。结论构建成功携带针对人Survivin的shRNA的肿瘤增殖型腺病毒载体Ad-delE1b55kD-shRNA/Survivin-EGFP,其能选择性地高效转染结肠癌细胞株HT-29并有效干扰Survivin,且较之增殖缺陷型腺病毒载体和脂质体载体具有更高的干扰效率。

References

[1]  王春毅, 傅仲学. Ad-delE1b55kD-shRNA/Survivin-EGFP的构建及鉴定[J]. 第三军医大学学报, 2007, 29(13):1293-1297.
[2]  范婷婷,唐良萏.Nek2基因沉默对卵巢癌SKOV3细胞侵袭能力的影响[J].第三军医大学学报,2012,34(15):1514.  Fan Tingting,Tang Liangdan.Silencing Nek2 via RNAi suppresses invasiveness in ovarian cancer SKOV3 cells[J].J Third Mil Med Univ,2012,34(13):1514. [2]梁佳,林力,寿铸,等.鼻咽癌畸胎瘤细胞源性生长因子表达及siRNA对HNE-1细胞株增殖的抑制[J].第三军医大学学报,2012,34(15):1572.  Liang Jia,Lin Li,Shou Zhu,et al.Expression of PCDGF in nasopharyngeal carcinoma and its silencing in cell proliferation[J].J Third Mil Med Univ,2012,34(13):1572. [3]刘丹,尹东,汤蕾,等.LPS预处理对心肌细胞缺氧/复氧损伤的保护作用[J].第三军医大学学报,2012,34(17):1707.  Liu Dan,Yin Dong,Tang Lei,et al.Protective effect of lipopolysaccharide pretreatment on cardiomyocytes anoxia/reoxygenation injury[J].J Third Mil Med Univ,2012,34(13):1707. [4]赵亮,高蕊,刘菁,等.异位表达FOXO4对胃癌细胞凋亡的影响[J].第三军医大学学报,2015,37(18):1830.  Zhao Liang,Gao Rui,Liu Jing,et al.Effects of ectopic expression of Forkhead box O4 on cell apoptosis in human gastric cancer cells[J].J Third Mil Med Univ,2015,37(13):1830. [5]黄士隋,史良会,黄广岩.RNA干扰下调LAT1表达对胃腺癌SGC-7901细胞增殖、侵袭、转移及细胞周期的影响[J].第三军医大学学报,2013,35(04):320.  Huang Shisui,Shi Lianghui,Huang Guangyan.Effect of RNA interference targeting LAT1 on proliferation, migration and invasion of SGC7901 cells[J].J Third Mil Med Univ,2013,35(13):320. [6]党微旗,唐浩,曹红,等.可调控STAT3干扰载体抑制BIU-87细胞侵袭的体外研究[J].第三军医大学学报,2013,35(05):400.  Dang Weiqi,Tang Hao,Cao Hong,et al.Effect of CRE-dependent RNA interference targeting STAT3 on invasion and migration in human bladder cancer BIU-87 cells[J].J Third Mil Med Univ,2013,35(13):400. [7]张彦,郑晓东,唐鹏,等.Sphk1基因对人乳腺癌MCF-7细胞增殖、凋亡和迁移能力的影响[J].第三军医大学学报,2012,34(21):2141.  Zhang Yan,Zheng Xiaodong,Tang Peng,et al.Sphk1 interference suppresses proliferation, apoptosis and migration in human MCF-7 breast cancer cells[J].J Third Mil Med Univ,2012,34(13):2141. [8]王勇,唐川,兰曦,等.沉默CD133基因对CD133+肝癌干细胞放射敏感性的影响[J].第三军医大学学报,2012,34(23):2373.  Wang Yong,Tang Chuan,Lan Xi,et al.RNAi targeting CD133 enhances radiosensitivity in CD133 positive liver cancer stem cells[J].J Third Mil Med Univ,2012,34(13):2373. [9]赵亮,李静,魏强,等.人PGI基因siRNA慢病毒质粒的构建及对白血病细胞增殖的影响[J].第三军医大学学报,2013,35(01):20.  Zhao Liang,Li Jing,Wei Qiang,et al.Construction of a lentiviral vector expressing human phosphoglucose isomerase gene siRNA and its influence on leukemia cell proliferation[J].J Third Mil Med Univ,2013,35(13):20. [10]熊震,汤旭东,房殿春,等.人肝素酶RNAi序列的筛选及鉴定[J].第三军医大学学报,2007,29(20):1929.  XIONG Zhen,TANG Xu-dong,FANG Dian-chun,et al.Screening and identification of heparanase RNAi sequence[J].J Third Mil Med Univ,2007,29(13):1929.

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