低浓度5-氟尿嘧啶对胃癌细胞生物学特性的影响

Effects of low level of 5-fluorouracil on biological characteristics of gastric cancer cell lines

目的探讨低浓度5-氟尿嘧啶(5-FU)对胃癌细胞转录因子OCT4、SOX2及细胞增殖能力的影响。方法用MTT法检测并确定5-FU对胃癌细胞系MGC803、SGC-7901的半数抑制浓度(IC50),实时荧光定量PCR及western blot检测经不同浓度的5-FU处理后胃癌细胞转录因子OCT4、SOX2表达水平,平板克隆形成实验分析胃癌细胞增殖能力的改变情况。结果5-FU对MGC803、SGC-7901细胞的IC50分别为(28.82±1.25)和(23.95±1.34);低浓度5-FU(0.5、1μmol/L)处理后,胃癌细胞MGC803、SGC-7901中转录因子OCT4、SOX2在mRNA及蛋白质水平中的表达均增强,差异具有统计学意义(P<0.05);平板克隆形成实验结果表明,经低浓度5-FU(0.5、1μmol/L)作用后胃癌细胞的增殖能力变强,形成的克隆数增多,差异有统计学意义(P<0.05);但5-FU浓度为2μmol/L时OCT4和SOX4的表达有不同程度的减弱,且克隆形成能力也有下降。结论低浓度5-FU能增强胃癌细胞转录因子表达、促进胃癌细胞增殖。

Objective To explore the effects of low level of 5-fluorouracil（ 5-FU） on biological characteristics of gastric cancer cell lines. Methods The half inhibition concentration（ IC50） of gastric cancer cells was detected by MTT assay. qRT-PCR and western blot were applied to determine the effects of different concentration of 5-FU level of transcription factor OCT4 and SOX2 expression in gastric cancer cells. Colony formation assay was used to analyze the growth ability of gastric cancer cells treated with low level of 5-FU.Results MTT assay showed that the IC50 of MGC803 and SGC-7901 were 28. 82 ± 1. 25 and 23. 95 ± 1. 34 respectively. qRT-PCR and western blot revealed that low level of 5-FU（ 0. 5μmol / L、1μmol / L） enhanced the stemness of gastric cancer cells. At mRNA and protein level,the expression of OCT4 and SOX2 were upregulated than control group in MGC803 and SGC-7901 cells,showing significant difference. Colony formation assay showed that low level of 5-FU（ 0. 5 μmol / L、1 μmol / L） could obviously promote gastric cancer cells growth. When the level of 5-FU to achieve 2 μmol / L,the expression of OCT4 and SOX2 were downregulated than control group in MGC803 and SGC-7901 cells,and colony formation assay showed inhibit gastric cancer cells growth. Conclusion Our results indicate that low level of 5-FU enhanced gastric cancer cells transcription factor expression,and promoted proliferation.