摘要
目的探讨下调胸苷酸合成酶（Ts）基因以及化疗药洛拉曲克对人结直肠癌LOVO细胞胸苷酸合成酶表达水平以及对细胞生长、凋亡的影响。方法构建针对人Ts基因的小分子干扰RNA（siRNA）和阴性对照，转染至LOVO细胞，利用RT—PCR和Westernblot技术观察沉默TS基因后其基因和蛋白表达水平的变化，MTF法检测细胞增殖情况；另联合洛拉曲克作用于LOVO细胞，观察两者对LOVO细胞的TS蛋白表达和细胞生长的影响。结果转染TSsiRNA后，LOVO细胞TSmRNA及蛋白的表达水平明显低于阴性对照组，细胞增殖速度亦低于对照组（均P〈0．05）。TSsiRNA联合洛拉曲克组细胞IC50值为（1．46±0．25）μm01／L，而阴性对照组和单用洛拉曲克组的IC50值分别为（6．81±0．31）μmol／L和（6．47±0．43）μmol／L。TSsiRNA联合洛拉曲克作用于细胞36h后。细胞凋亡指数为（62．12±0．89）％，高于单用TSsiRNA和洛拉曲克[（21．56±0．67）％和（40．51±0．83）％．均P〈0．05]。结论TSsiRNA可以下调人LOVO细胞中TS基因和蛋白的表达，使细胞生长速度减慢，凋亡增加，并可以增强洛拉曲克的药物敏感性。
Objective To investigate the effect of the RNAi and the chemotherapy drugs nolatrxed on the expression of thymidylate synthase （TS） and the growth of the colorectal carcinoma LOVO cells. Methods The siRNA was constructed targeting the human TS gene, and then transfected into the human colorectal cancer LOVO cells. RT-PCR and Western blot technique were used to observe the TS gene and protein expression levels, and MTT was used to detect cell proliferation after silencing the TS gene. In addition, siRNA and nolatrxed were applied to the LOVO cells to observe the TS protein expression and cell growth. Results TS siRNA significantly reduced the expression of TS gene and protein in LOVO cells, and inhibited cell growth. The IC50 value of LOVO cells was （1.46±0.25） μmol/L in TS siRNA combined with nolatrexed group, （6.81±0.31） μ mol/L in the negative control group, and （6.47±0.43） μmol/L in the single nolatrexed group. After treatment of TS siRNA combined with nolatrexed on LOVO cells for 36 hours, the apoptosis index was higher than that in single TS siRNA and nolatrexed [（62.12±0.89）% vs. （21.56±0.67）% and （40.51±0.83）%, both P〈0.05]. Conclusion TS siRNA can partly suppress the expression of TS gene in LOVO cells, inhibit cell proliferation, promote cell apoptosis and enhance cell sensitivity to apoptosis induced by nolatrexd.
出处
《中华胃肠外科杂志》
CAS
2012年第11期1187-1191,共5页
Chinese Journal of Gastrointestinal Surgery
基金
山东省医药卫生科技发展计划项目（2007QZ015）
