黄连素联合胞嘧碇脱氨酶自杀基因对直肠癌细胞的抑制作用

目的 探讨黄连素联合胞嘧碇脱氨酶自杀基因(Ad-CD)对直肠腺癌细胞的体外杀伤作用.方法 培养人直肠腺癌细胞株HR-8348细胞,将重组腺病毒自杀基因转染人直肠癌细胞株HR-8348后,用四甲基偶氮唑盐(MTT法)检测黄连素联合Ad-CD基因对HR-8348细胞存活率的影响.结果 构建pcDNA3.1( )-CD真核表达载体成功,CD基因序列分析表明:重组子pcDNA3.1( )-CD含有完整的CD基因.RT-PCR表明CD基因在HR-8348细胞中表达,成功筛选出稳定表达CD基因的HR-8348细胞.HR-8348-CD细胞对浓度为100μg/ml以上的5-FC敏感,随着药物浓度升高和作用时间的延长,杀伤作用增强,但在250μg/ml以上浓度变化时,细胞杀伤率变化不大.分别以0.1、0.3、3.0、30.0μm浓度的黄连素联合5-FC对直肠癌细胞作用,细胞抑制作用随黄连素浓度升高而上升,但3.0、30μm浓度作用变化不大.在250μg/ml 5-FC浓度下,0.1、0.3、3.0、30.0μm浓度的黄连素联合5-FC对直肠癌细胞生长抑制率分别为:27.7%、42.4%、52.3%、56.3%.结论 黄连素联合5-Fc作用HR-8348-CD细胞能够增强对直肠癌细胞的抑制作用.

Inhibition by Berberine and Cytosine Deaminase Enzyme Suicide Gene on Rectum Cancer Cells.

Objective To investigate the killing effect by berberine and cytosine deaminase enzyme suicide gene on rectum cancer cells in vitro.Methods Human rectum cancer HR-8348 cells were cultured,then transfected by recombinant adenovirus suicide gene the survivial percentage of which was examined by MTT method.Results Recon of pcDNA3.1(+)-CD was constructed successfully.The CD gene sequence analysis indicated th a+ recon pcDNA3.1(+)-CD contain intact gene.RT-PCR showed CD gene expressed stably in the HR-8348 cells...