黄芪多糖联合5-氟尿嘧啶对骨肉瘤细胞系MG63细胞中CD133^+细胞凋亡影响及其分子机制  

Effect of astragalus polysaccharide combined with 5-FU on apoptosis of CD133^+ cells in osteosarcoma cell line MG63 and its molecular mechanism

作　　者：唐晓栋[1] TANG Xiao -dong(Spinal Surgery Department I, Gansu Hospital of Traditional Chinese Medicine,Lanzhou,Gansu 730000,China)

机构地区：[1]甘肃省中医院脊柱一科

出　　处：《中国卫生检验杂志》2018年第23期2825-2829,共5页Chinese Journal of Health Laboratory Technology

摘　　要：目的探索黄芪多糖联合5-氟尿嘧啶(5-FU)对骨肉瘤细胞系MG63细胞中CD133^+细胞凋亡的影响及分子机制。方法免疫磁珠分选MG63细胞中CD133^+细胞; MTT实验检测黄芪多糖、5-FU以及联合使用对CD133^+细胞增殖的影响;线粒体膜电位检测试剂盒检测细胞中线粒体膜电位; q PCR实验检测细胞中Bax、Bcl-2 mRNA表达;Western blot实验检测细胞中Cyt C、Caspase-3、Caspase-9蛋白表达;体内观察干预后CD133^+移植瘤的生长; Western blot实验检测移植瘤中Caspase-3、Caspase-9蛋白表达。结果磁珠分选富集的CD133^+细胞,纯度达到90%。经干预后发现与单独使用相比,联合使用对CD133^+细胞和CD133^+细胞移植瘤增殖生长具有更强的抑制,线粒体膜电位显著降低,同时诱导CD133^+细胞中Bax mRNA、Cyt C蛋白以及体内外细胞中Caspase-3、Caspase-9蛋白表达显著升高,显著抑制Bcl-2 mRNA表达。结论与单独干预相比,黄芪多糖联合5-FU可显著诱导MG63细胞中CD133^+细胞及其移植瘤凋亡,其分子机制可能与线粒体凋亡相关。Objective To explore the effect and mechanism of astragalus polysaccharide combined with 5-FU on apoptosis of CD133^+ cells in osteosarcoma cell line MG63. Methods Immunomagnetic beads sorting was used to separate CD133^+ cells from MG63 cells. MTT assay was used to detect the effects of astragalus polysaccharide,5-FU and combined use on proliferation of CD133^+ cells. Mitochondrial membrane potential detection kit was used to detect mitochondrial membrane potential in cells; q PCR assay was used to detect the expression of Bax and Bcl-2 mRNA in cells; Western blot was used to detect the levels of Cyt C,Caspase-3 and Caspase-9 in cells; the growth of CD133^+ transplanted tumor was observed in vivo; Western blot was used to detect the levels of Caspase-3 and Caspase-9 protein in transplanted tumors. Results Magnetic beads were sorted and enriched in CD133^+ cells with a purity of 90%. After intervention,the combined use had stronger inhibition on the proliferation of CD133^+ cells and CD133^+ cells,and resulted in lower mitochondrial membrane potential and induction of CD133^+ cells in vitro. The levels of Caspase-3 and Caspase-9 in Bax mRNA,Cyt C protein and in vitro and in vivo cells significantly increased,which significantly inhibited the level of Bcl-2 mRNA. Conclusion Compared with the single intervention,astragalus polysaccharide combined with 5-FU can significantly induce apoptosis of CD133^+ cells and their transplanted tumors in MG63 cells,and its molecular mechanism may be related to mitochondrial apoptosis.

关 键 词：骨肉瘤 黄芪多糖 5-氟尿嘧啶 联合 凋亡 

分 类 号：R738.1[医药卫生—肿瘤]