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

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

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/ml5-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。

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, which is sensitive to 5 - FC above the density of 100ug/ml. Elevated along with the medicine density with response time lengthening, its damaging effect enhanced. However for the density above 5-FC 250g/ml, the change of cell killing rate was not apparent, when by 0.1 , 0.3, 3.0, 30.0um density berberine unites the 5 - FC on rectum cancer of the intestines Respectively, the cell inhibitory action elevates along with density rises, but 3. 0um, 30um the inhibition change was not apparent. Under 250ug/ml of 5 - FC, 0. 1, 0. 3, 3. 0, 30. 0um of berberine united the 5 - FC The HR -8348 cells growth suppression rate were： 27.7% ,42.4% , 52.3% , 56.3% respectively. Conclusion Inhibition function to human rectum cancer HR -8348 cells can be strengthened by berberine combined 5 - FC.