摘要
目的研究胃癌侧群(Side Population,SP)细胞对化疗药物5-FU(氟尿嘧啶)的耐药性及可能机制,并检测干细胞相关基因Nanog、Musashi-1及CD44的表达情况。方法选择人胃癌细胞株SGC-7901,以荧光染料Hoechst 33342染色,维拉帕米拮抗对照,应用流式细胞仪分选SP细胞和nonSP细胞。细胞耐药实验比较SP细胞与nonSP细胞对化疗药物5-FU的耐药性差异;western-blot检测ABCG2和bcl-2蛋白表达情况;流式细胞仪分析细胞周期;荧光定量PCR检测两组细胞中干细胞相关基因Nanog、Musashi-1及CD44 mRNA的表达差异。结果胃癌细胞株SGC-7901中SP细胞的比例为2.8%,SP细胞对5-FU的耐药存活率明显高于nonSP细胞(P<0.05),与nonSP细胞相比,SP细胞高表达耐药蛋白ABCG2和抗凋亡蛋白bcl-2,有更多的细胞处于G0/G1期(P<0.05),并高表达干细胞相关基因Musashi-1和CD44。结论胃癌SGC-7901细胞株中SP细胞对化疗药物5-FU的耐药性明显高于nonSP细胞,其耐药机制可能与SP细胞高表达耐药蛋白ABCG2和抗凋亡蛋白bcl-2,有更多细胞处于G0/G1期有关;Musashi-1和CD44可能是相对特异性的胃癌干细胞标志物。
Objective to research chemoresistanee of SP cells to 5-FU and examine stem ceil related genes of SP cells from gastric cancer SGC- 7901 cell strain. Methods SGC-7901 cells were sorted as SP cells and nonSP cells using flow eytometry after SGC-7901 cells were stained with Hoechst33342 ;chemoresistance experiment was used to observe two group cells＇ resistance to 5-FU ;western-blot was performed to examine the expression of ABCG2 and Bel-2;flow cytometry was applied to analyze eeil cycle;the mRNA expression of Nanog, Musashi-1 and CD44 was detected by real-time PCR. Results SP cells have a stronger resistance to 5-FU and have a higher expression of ABCG2 and Bcl- 2 than nnnSP ceils,and the proportion of cells in GO/G1 phase is also larger than nonSP cells;SP cells show a higher expression of Musashi-l and CD44 than nonSP cells. Conclusion SP cells have a higher resistance to 5-FU than nonSP cells, the possible mechanisms are related to higher expression of ABCG2 and Bel2 and more cells in G0/G1 phase. ; Musashi-l and CD44 may be the markers of gastric cancer stem cells.
出处
《世界科技研究与发展》
CSCD
2011年第6期1081-1084,共4页
World Sci-Tech R&D
