摘要
目的探讨氨基硅烷化Fe3O4磁性纳米介导的pcDNA3.1(+)-p53质粒(-p53质粒)对人肝癌HepG2细胞的作用。方法(1)以-p53质粒作为表达基因,用氨基硅烷化Fe3O4磁性纳米粒进行细胞转染,计算转染率,并与脂质体转染组,空白载体组及空白对照组进行比较。(2)观察在外加磁场条件下的氨基硅烷化Fe3O4磁性纳米粒的转染效率和速度。(3)观察-p53磁性纳米粒对HepG2细胞生长增殖的抑制作用。结果(1)RT-PCR及Westernblot检测证实,载有氨基硅烷化Fe3O4的磁性纳米粒能成功的在人肝癌HepG2细胞中导入了外源性的野生型p53基因,其转运效率(39%)略高于相同条件下脂质体的转染效率(36%)(P>0.05)。(2)在外加磁场条件下氨基硅烷化Fe3O4磁性纳米粒的转染效率明显增强。(3)转染的外源性p53基因可明显抑制人肝癌HepG2细胞的生长速度及集落形成能力,使细胞阻滞于G1期,抑制率为46%。结论(1)氨基硅烷化Fe3O4纳米颗粒是良好的基因转运载体,可将外源性基因成功的转入靶细胞。(2)转入外源性野生型p53能有效抑制HepG2细胞的生长增殖。该研究为基因治疗恶性肿瘤的在体实验奠定了基础。
Objective To explore the effect of amino-functional Fe3O4 nanoparticles-mediated pcDNA3. 1 （ ＋ ） - p53 （ - p53 ） on human hepatoma cell line-HepG2 cell. Methods （ 1 ） Use of functional Fe3O4 nanoparticle as a vector to transfect-p53 into HepG2 cells, calulate the transfected rate, and compare it with lipofectamine vector group, blank functional Fe3O4 nanoparticles （FFN） group and control group. （2） Observation of the trasfected rate and velocity of FFN under addition of extra-magnetic field. （ 3 ） Observation of the inhibition effect of trasfected-p53 on HepG2 cells growth. Results （ 1 ） RT-PCR and Western blot examination comfirmed that FFN could successfully transfect-p53 into HepG2 cells, the transfection rate （39%） was slightly higher than that of liposome （36%） （P 〉 0. 05 ）. （2） The transfection rate and velocity could be increased by addition of extra-magnetic field. （3） Transfected-p53 obviously inhibited the growth of HepG2 at G1 phase; the growth inhibition rate of HepG2 was 46%. Conclusions （ 1 ） FFN is a good vector for gene transfection, it can successfully transfect the gene into target cells. （2） Transefected- p53 can effectively inhibit the growth of HepG2. This result can establish a foundation for animal experimental research of gene therapy of malignancy.
出处
《中国普通外科杂志》
CAS
CSCD
2008年第6期578-583,共6页
China Journal of General Surgery
