FasL cDNA转染对直肠癌细胞顺铂耐药性的影响

目的:探讨FasLcDNA转染和表达对直肠癌细胞耐药性的影响。方法:用RT-PCR方法克隆人FasL全长cDNA,构建pcDNA3.1-FasL真核表达载体,用脂质体法转染HR-8348人直肠癌细胞,采用MTT法检测顺铂对转染和未转染直肠癌细胞的生长抑制率。结果:DNA测序证实克隆FasLcDNA898bp与GeneBank序列完全一致。构建真核表达载体转染HR-8348细胞后,FasLmRNA表达明显增强。在不同浓度顺铂(1、5、10、20、40mg/L)的作用下,FasL转染组直肠癌细胞抑制率分别为11.0%、25.4%、31.2%、37.8%、42.4%:对照组癌细胞抑制率分别为26.1%、34.4%、37.6%、42.9%、53.2%,其差异有显著性意义(t=4.43,P<0.05)。结论:FasL转染HR-8348细胞可增强癌细胞的耐药性,减弱顺铂对HR-8348细胞的杀伤作用。

FasL cDNA transfection alters rectal cancer cell tolerance to cisplatin

Objective To study the effect of FasL cDNA transfection and expression on rectal cancer cell tolerance to cisplatin.Methods RT-PCR was used to clone full length of FasL cDNA from active peripheral mononuclcar cells of healthy donors.An eukaryotic expressing vector pcDNA3.1-FasL was constructed and transfected into HR-8348 human rectal cancer cell line with lipofectin. The growth suppression rate of transfected and non-transfected cancer cells were tested with MTT method. Results A FasL cDNA of 898bp was obtained and the sequence was compared with that in GeneBank. Enhanced expression of FasL mRNA were found in cancer cells with recombinant vector transfection. When the concentration of cisplatin was 1, 5, 10, 20 and 40 mg/L respectively, the suppression rate of cancer cells in the FasL transfected group as 11.0%, 25.4%, 31.2%, 37.8% and 42.4% respectively, and in the non-transfected group, the suppression rate was 26.1%, 34.4%, 37.6%, 42.9% and 53.2% respectively. The difference was significant between the transfection and non-transfection groups(t=4.43,P<0.05). Conclusions FasL cDNA transfection and enhanced FasL expression could strengthen the tolerance of cancer cells to cisplatin and weaken its chemotherapeutic effect.