目的研究甲基化寡核苷酸灭活Dickkopf相关蛋白3(Dickkopf-related protein 3,DKK3)基因对Hep G2肝癌细胞增殖的影响。方法采用甲基化特异性PCR法检测肝癌细胞DKK3基因启动子甲基化状态。将寡核苷酸(分别为MON组、UMON组、CON1组和CON2组...目的研究甲基化寡核苷酸灭活Dickkopf相关蛋白3(Dickkopf-related protein 3,DKK3)基因对Hep G2肝癌细胞增殖的影响。方法采用甲基化特异性PCR法检测肝癌细胞DKK3基因启动子甲基化状态。将寡核苷酸(分别为MON组、UMON组、CON1组和CON2组)转染Hep G2肝癌细胞,比较转染后DKK3基因启动子甲基化状态、细胞增殖和凋亡的差异。结果 Hep G2肝癌细胞DKK3基因启动子未检测到甲基化,Hep3B、SMMC-7721和Hu H-7肝癌细胞DKK3基因启动子处于甲基化状态。MON可成功诱导其互补序列CG位点产生甲基化,而UMON、CON1和CON2寡核苷酸无法诱导甲基化。UMON组、CON1组和CON2组Hep G2肝癌细胞在D0-D6吸光度、G_0/G_1期、S期、G_2/M期、增殖指数(proliferation index,PI)和凋亡率方面差异均无统计学意义(均P>0.05);MON组Hep G2肝癌细胞G_0/G_1期比例和凋亡率均显著低于UMON组、CON1组和CON2组(均P<0.05),而D1-D6吸光度、S期、G_2/M期和PI均显著高于UMON组、CON1组和CON2组。结论甲基化寡核苷酸可成功诱导DKK3基因甲基化,导致Hep G2肝癌细胞细胞增殖增加而凋亡下降。展开更多
Functionalized carbon nanotubes （CNTs） were made for the delivery of genes and drugs and CNT-based biosensors. The basis of CNTs is for binding with biomolecules in biomedical applications. The binding tendency with...Functionalized carbon nanotubes （CNTs） were made for the delivery of genes and drugs and CNT-based biosensors. The basis of CNTs is for binding with biomolecules in biomedical applications. The binding tendency with small interfering RNA oligonucleotides and cytotoxicity of cetyltrimethyl ammonium bromide （CTAB）-coated single-walled carbon nanotubes （SWNTs） were studied. The field emission scanning electron microscopy and transmission electron microscopy results show that a SWNT suspension in CTAB solution was well-dispersed and stable. CTAB is the cross-linker between SWNTs and oligonucleotides. The CTAB-coated SWNTs have less cytotoxicity to human umbilical vein endothelial cells than single SWNTs and the cytotoxicity of CTAB-coated SWNTs depended on the concentration of CTAB-coated SWNTs.展开更多
基金Project (30770838) supported by the National Natural Science Foundation of China Project (2008WK2003) supported by Hunan Science and Technology Foundation, China+1 种基金 Project (2010QZZD006) supported by the Key Program of Central South University Advancing Front Foundation, China Project (200806) supported by Opening Foundation of State Key Laboratory of Powder Metallurgy, Central South University, China
文摘Functionalized carbon nanotubes （CNTs） were made for the delivery of genes and drugs and CNT-based biosensors. The basis of CNTs is for binding with biomolecules in biomedical applications. The binding tendency with small interfering RNA oligonucleotides and cytotoxicity of cetyltrimethyl ammonium bromide （CTAB）-coated single-walled carbon nanotubes （SWNTs） were studied. The field emission scanning electron microscopy and transmission electron microscopy results show that a SWNT suspension in CTAB solution was well-dispersed and stable. CTAB is the cross-linker between SWNTs and oligonucleotides. The CTAB-coated SWNTs have less cytotoxicity to human umbilical vein endothelial cells than single SWNTs and the cytotoxicity of CTAB-coated SWNTs depended on the concentration of CTAB-coated SWNTs.
