Tian Shu-bo, Jing Chang-qing, Li Le-ping
Department of Gastrointestinal Surgery, Provincial Hospital, Shandong University, Jinan 250021, China.
Zhonghua Wei Chang Wai Ke Za Zhi. 2012 Nov;15(11):1187-91.
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.
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.
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) μmol/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].
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.
探讨RNA干扰（RNAi）及化疗药物诺拉曲塞对胸苷酸合成酶（TS）表达及大肠癌细胞株LOVO生长的影响。
构建针对人TS基因的小干扰RNA（siRNA），转染人结肠癌LOVO细胞。采用逆转录-聚合酶链反应（RT-PCR）及蛋白质免疫印迹法（Western blot）观察TS基因及蛋白表达水平，噻唑蓝（MTT）法检测TS基因沉默后细胞增殖情况。此外，将siRNA与诺拉曲塞作用于LOVO细胞，观察TS蛋白表达及细胞生长情况。
TS siRNA可显著降低LOVO细胞中TS基因及蛋白表达，抑制细胞生长。TS siRNA联合诺拉曲塞组LOVO细胞的半数抑制浓度（IC50）值为（1.46±0.25）μmol/L，阴性对照组为（6.81±0.31）μmol/L，单纯诺拉曲塞组为（6.47±0.43）μmol/L。TS siRNA联合诺拉曲塞作用于LOVO细胞36小时后，凋亡指数高于单纯TS siRNA组及单纯诺拉曲塞组[（62.12±0.89）%比（21.56±0.67）%和（40.51±0.83）%，P均<0.05]。
TS siRNA可部分抑制LOVO细胞中TS基因表达，抑制细胞增殖，促进细胞凋亡，并增强细胞对诺拉曲塞诱导凋亡的敏感性。
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