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作 者:钟婧[1] 黄惠莲[1] 李静[1] 钱福初[1] 李丽琴[1] 戴利成[1]
机构地区:[1]湖州市中心医院分子医学重点实验室,浙江313000
出 处:《中华实验外科杂志》2015年第10期2402-2404,共3页Chinese Journal of Experimental Surgery
基 金:浙江省自然科学基金资助项目(Y2111250);湖州市科技攻关计划资助项目(2011GG14)
摘 要:目的合成一种新的基因递送载体一氨基化壳聚糖,观察氨基化壳聚糖一中期因子反义寡核苷酸(MK-ASODN)对肝癌细胞生长的影响。方法使用氨基化壳聚糖包被MK-ASODN制备纳米粒,用纳米粒[每孔含MK-ASODN150pmol,氨基化壳聚糖:MK-ASODN(质量比=16:1、32:1、64:1)]转染肝癌细胞HepG2,通过实时荧光定量聚合酶链反应(FQ-PCR)检测MK基因沉默情况;并通过四唑氮化合物(MTS)实验观察纳米粒[各组别分别含blK-ASODN2.5,5.0、7.5、10.0、12.5pmol,氨基化壳聚糖:MK-ASODN(质量比)=64:1]对HepG2生长的影响。结果氨基化壳聚糖-MK-ASODN纳米粒可抑制HepG2细胞中MK基因mRNA的表达,基因沉默率最高可达到(66.57±1.1)%,并由此可抑制HepG2细胞的生长,细胞生长抑制率可达(73.48±1.00)%。结论通过氨基化壳聚糖介导MK-ASODN进人HepG2细胞可抑制其生长。Objective To investigate the inhibitory effects of midkine antisense oligonucleotide (MK - ASODN) aminated chitosan nanoparticles on hepatocellular carcinoma growth. Methods The HepG2 cells were transfected with MK- ASODN aminated chitosan nanoparticles [ 150 pmol/well, amina- ted chitosan: MK- ASODN (w:w) = 16:1, 32:1, 64:1 ], and the efficiency of MK gene silence was de- tected by real-time fluorescent quantitative polymerase chain reaction ( FQ - PCR). The suppression of HepG2 growth by MK - ASODN aminated chitosan nanoparticles [ MK - ASODN 2. 5, 5.0, 7.5, 10. 0, 12. 5 pmol/well, aminated chitosan : MK - ASODN ( w: w) = 64 : 1 ] was evaluated using muhitetrazolium compounds (MTS) assay. Results The mRNA expression of MK gene in HepG2 could be down - regula- ted by MK - ASODN aminated chitosan nanoparticles with the highest gene silence rate being (66. 57 ±1.1 ) %, and the HepG2 growth could be suppressed by MK - ASODN arninated chitosan nanoparticles with the suppression rate being (73.48 ± 1.00) %. Conclusion The MK - ASODN aminated chitosan nanopar- ticles could be transfected into HepG2 ceils and suppressed the HepG2 growth.
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