胸苷酸合成酶mRNA在结肠癌HT-29细胞中表达的意义及与HT-29细胞对雷替曲塞化疗敏感性的相关性研究

Significance of thymidylate synthase mRNA expression in colon cancer HT-29 cells and its correlation with HT-29 cells sensitivity to raltitrexed chemotherapy

目的探讨胸苷酸合成酶(TS)mRNA在结肠癌HT-29细胞中表达的意义,并探讨其与HT-29细胞对雷替曲塞化疗敏感性的相关性。方法对数期生长HT-29细胞,将TS-pcDNA3.1-zeo(+)质粒及pcDNA3.1 empty vector质粒转染至HT-29细胞,分别命名为过表达组、空载组,不干预细胞命名为对照组;实时荧光定量-聚合酶链反应法检测3组TS mRNA表达情况,MTT试验检测细胞增殖情况及计算雷替曲塞对各组细胞的半数抑制浓度(IC50)。结果过表达组,对照组和空载组细胞TS mRNA相对表达量分别为0.85±0.16,0.13±0.03,0.14±0.03,过表达组高于对照组和空载组,差异有统计学意义(均P<0.001)。过表达组和空载组干预48 h时增殖抑制率分别为(48.38±3.26)%,(1.27±0.14)%,过表达组增殖抑制率高于空载组(P<0.05)。雷替曲塞对过表达组,对照组和空载组中细胞的IC50值分别为(20.60±1.95),(43.51±4.11),(43.56±4.28)μmol·L^-1,雷替曲塞对过表达组细胞的IC50值低于对照组和空载组,差异均有统计学意义(均P<0.05)。结论TS mRNA低表达可促进结肠癌HT-29细胞增殖,降低细胞对雷替曲塞敏感性,过表达TS基因可抑制细胞增殖,提高对雷替曲塞敏感性。

Objective To investigate the significance of thymidylate synthase(TS)mRNA expression in colon cancer HT-29 cells,and to explore its correlation with HT-29 cells sensitivity to raltitrexed chemotherapy.Methods The TS-pcdna3.1-zeo(+)and pcDNA3.1 empty vector plasmids were transfected into HT-29 cells,which were set as over expression group and no-load group.The control group was non intervention cells.The TS mRNA expression in three groups were detected by real-time fluorescence quantitative PCR.The cell proliferation and calculate the half inhibitory concentration(IC50)of letetrexed were detected by MTT assay.Results The relative expression of TS mRNA in over expression group,control group and no-load group was 0.85±0.16,0.13±0.03 and 0.14±0.03,respectively.The relative expression of TS mRNA in over expression group was higher than that in control group and no-load group(all P<0.001).The proliferation inhibition rates were 48.38±3.26%and 1.27±0.14%in the overexpression group and the no-load group respectively.The proliferation inhibition rates in overexpression group was higher than that in the no-load group(P<0.05).The IC-50 values of letetrexed in the over expression group,control group and no-load group were 20.60±1.95μmol·L^-1,43.51±4.11μmol·L^-1 and 43.56±4.28μmol·L^-1,respectively.The IC50 value of letetrexed group was lower than that of control group and no-load group(all P<0.05).Conclusion Low expression of TS mRNA can promote the proliferation of colon cancer HT-29 cells and decrease the sensitivity of cells to raltitrex.Overexpression of TS gene can inhibit cell proliferation and increase sensitivity to raltitrex.