5-氟尿嘧啶及长春新碱对胃癌细胞SGC-7901干细胞相关特性的影响

Effects of 5-FU and vincristine on stem cell-associated phenotypes and properties of gastric cancer cell line SGC-7901

目的探讨化疗药物5-FU、长春新碱对胃癌细胞SGC-7901干细胞相关表型及功能影响。方法 5-FU、长春新碱分别作用于SGC-7901细胞后,通过流式细胞术及荧光实时定量PCR的方法观察胃癌干细胞候选标志CD44、CD133的表达,检测干性相关基因及上皮-间质转化(epithelial-to-mesenchymal transition,EMT)相关基因的表达变化,并检测药物作用对其集落形成能力、无血清培养基中成球能力及NOD/SCID小鼠体内成瘤能力的影响。结果不同浓度5-FU、长春新碱作用3 d后均可显著增加剩余SGC-7901细胞CD44 mRNA的总体表达水平,分别为未用药组的(2.36±0.03)、(1.74±0.05)、(6.03±0.75)、(4.06±0.77)倍,均有显著差异(P<0.05)。CD133及CD44阳性细胞比例亦明显升高,但干性基因表达、克隆形成能力、成瘤能力未见明显增加。而与5-FU不同的是,长春新碱作用后细胞发生了EMT相关基因的表达改变,在无血清培养基中形态与其他组细胞差异显著,且可形成典型的细胞球。结论 5-FU及长春新碱作用后细胞CD44和CD133的表达增加,此外长春新碱还促进细胞EMT改变,并增加细胞在无血清培养基中的成球能力。

Objective To identify the effects ofS-fluorouracil （5-FU） and vincristine （VCR） on stem cell-associated phenotypes and properties of gastric cancer cell line SGC-7901. Methods SGC-7901 cells were treated with 5-FU and VCR, respectively. Candidate stem cell markers of CD44 and CD133 as well as oth- er sternness genes and epithelial-to-mesenchymal transition （EMT）-associated genes were assayed. Colony for- mation assay, sphere formation assay and tumorigenic formation assay were used to test the properties of SGC- 7901 cells associated with cancer stem cells. Results After 5-FU and VCR treatment, the mRNA expression levels of CD44 and CD133 significantly increased in all the doses as compared with the control group （ up to 2.36± 0.03, 1.74± 0.05,6.03 ＋ 0.75 and 4.06 ±0. 77 folds respectively, P 〈 0. 05 ）, but the capabilities of colony formation and tumorigenic formation had no change. VCR treatment manifested the EMT gene profile and showed different morphologic features in serum-free SGC-7901 cell culture, which was different from 5-FU treatment. Conclusion Transient treatment of 5-FU and VCR up-regulates the expression of CD44 and CD133. VCR induces EMT changes in SGC-7901 cells and enhances sphere formation in serum-free culture medium.