洛拉曲克对人大肠癌LoVo细胞胸苷酸合酶水平及细胞增殖的影响

Effects of nolatrexed on thymidylate synthase expression and cell proliferation in hunman colorectal carcinoma LoVo cells

目的:研究洛拉曲克体外对人大肠癌LoVo细胞胸苷酸合酶(TS)水平动态变化以及对细胞增殖的影响。方法:采用四甲基偶氮唑蓝(MTT)法检测不同浓度的洛拉曲克对LoVo细胞抑制率的影响,RT-PCR和Wes tern blot法检测TS基因和蛋白的表达变化,流式细胞术检测细胞凋亡率的变化。结果:洛拉曲克对LoVo细胞有明显的抑制作用,其IC50值约为6.31μmol/L;并可促进细胞凋亡,凋亡率随作用时间的延长而增加。RT-PCR结果显示,在mRNA水平,相同剂量洛拉曲克处理细胞后,随着时间的延长,TS/GAPDH比值与对照组相比,有持续升高的趋势,但是只有48 h和60 h相对于未处理组差异有统计学意义。相同剂量的洛拉曲克处理细胞后,随着时间的延长,其TS蛋白的表达水平亦有持续升高的趋势。结论:洛拉曲克可以明显促进大肠癌细胞的凋亡并可产生TS诱导现象,术后以此可以更好地指导临床用药。

Objective： To investigate the dynamic effect of thymidylate synthase in human colorectal carcinoma LoVo cells after treatment with nolatrexed.Methods： Use the MTT method to detect the inhibition ratio after treatment with different nolatrexed concentrations in LoVo cells,RT-PCR and Western blot technique to observe the TS gene and protein expression levels,flow cytometry to detect the apoptosis rate of LoVo cells.Results： Nolatrexed can inhibit the LoVo cells proliferation significantly,the IC50 is about 6.31μmol/L.It can also promote the cells apoptosis,the apoptosis rate increased with the action time extend.The result of RT-PCR demonstrated that in the mRNA levels,TS/GAPDH ratio continue to rise all along within the time period in LoVo cells,only the ratio of 48h and 60h have significant difference to the untreatment group.But,when LoVo cells were treated with the same dose in the different time,the level of TS protein expression also keep rising as time goes on.Conclusion： Nolatrexed can significantly promote cell apoptosis and produce TS-induced phenomenon,which can better guide clinical durg use.