论文部分内容阅读
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 investigate the effects of chemotherapy drugs 5-FU and vincristine on the phenotype and function of gastric cancer SGC-7901 stem cells. Methods After 5-FU and vincristine were applied to SGC-7901 cells respectively, the expression of CD44 and CD133, a marker of gastric cancer stem cells, was observed by flow cytometry and real-time quantitative PCR. The expressions of dry-related genes and epithelial-mesenchymal stem cells (EMT). The effect of drug on its colony-forming ability, ability of spheroidizing in serum-free medium and tumorigenicity of NOD / SCID mice were also investigated. Results The expressions of CD44 mRNA in the remaining SGC-7901 cells were significantly increased after treated with different concentrations of 5-FU and vincristine for 3 days (2.36 ± 0.03 and 1.74 ± 0.05, ± 0.75), (4.06 ± 0.77) times, all of which were significantly different (P <0.05). CD133 and CD44 positive cells also significantly increased, but no significant increase in the expression of dry gene, clonality, tumorigenicity. However, unlike 5-FU, the expression of EMT-related genes was changed after vincristine treatment. The morphology of EMT-related genes in serum-free medium was significantly different from that of other cells, and typical cell spheres were formed. CONCLUSION: The expression of CD44 and CD133 increased after 5-FU and vincristine treatment. In addition, vincristine also promoted the change of EMT and increased the ability of spheroid formation in serum-free medium.