bFGF对鼻咽癌干细胞体外培养增殖及SCF表达的影响

Effects of bFGF on proliferation and the expression of SCF in cultured nasopharyngeal carcinoma cells in vitro

目的:观察不同浓度碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)对体外培养鼻咽癌(nasopharyngeal carcinaoma,NPC)干细胞增殖的影响,及干细胞因子(stem cell factor,SCF)表达的变化。方法:以免疫磁珠技术从NPC细胞株CNE2中分选CD133+干细胞。分别以5,10,20ng/ml bFGF作用于NPC干细胞,MTT比色法和平板克隆形成实验测定CD133+细胞的体外增殖能力。ELISA检测各组细胞分泌SCF的含量;RT-PCR及Western-blot检测细胞中SCF的表达。结果:不同浓度bFGF作用于NPC干细胞,可促进干细胞的克隆形成,当bFGF作用第7天时干细胞增殖达峰值(P<0.05),呈剂量依赖性;同时分泌SCF含量显著增加,SCF mRNA及蛋白表达均显著上调(P<0.05),且呈剂量依赖特征。结论:bFGF可促进NPC干细胞增殖,其作用机制可能与促进SCF分泌、上调SCF的表达密切相关。

Objective：To observe the effects of various concentrations bFGF on proliferation and the expression of SCF in cultured nasopharyngeal carcinoma cells in vitro. Methods ：Immunomagnetic beads was used to sort CD133＋ stem cells from NPC cell line CNE2.5,10,20ng/ml bFGF were used to intervene NPC stem cells. The proliferation a- bility of CD133＋ in vitro was determined by MTI＇ colorimetric method and colony formation assay. The quantities of SCF were measured by ELISA in each group. The expression of SCF was determined by RT - PCR and Western - blot. Results：Various concentrations bFGF could promote the formation of cloning stem ceils. The proliferation of CD133＋ peaked at the seven day. They showed a dose - dependent characteristics. The quantities of SCF secretion of SCF increased by various concentrations bFGF acting on NPC stem cells （P 〈 O. 05 ）. So did the mRNA and protein expression. Conclusion： bFGF can promote the stem cell proliferation of NPC. And the mechanism may has a close re- lationship with promotion of SCF secretion and up - regulation of SCF expression.