不同浓度葡萄糖对sf9细胞生长特性及蛋白表达的影响  

Effects of different glucose concentrations on growth characteristics of and protein expression in sf9 cell

作　　者：丁军颖[1] 张振龙[1] 李军[1] 赵建英[1] 于世荣[1] 许雪梅[2] 

机构地区：[1]北京生物制品研究所中试一室,100024 [2]北京协和医学院基础学院,100730

出　　处：《国际生物制品学杂志》2010年第1期6-8,共3页International Journal of Biologicals

基　　金：国家高技术研究发展计划（2007AA022475）

摘　　要：目的研究不同浓度葡萄糖对昆虫细胞sf9生长特性及蛋白表达的影响。方法将对数生长期的s丹细胞在含不同浓度葡萄糖的Hyclone无血清培养基（serum—free medium，SFM）中培养。采用台盼蓝拒染法监测细胞密度，分析比较不同浓度葡萄糖对s固细胞生长特性的影响。然后，将对数生长期的sf9细胞在含不同浓度葡萄糖的HycloneSFM中培养表达人乳头瘤病毒（humanpa pillomavims，HPV）18型L1蛋白。以蛋白印迹法鉴定并分析比较不同浓度葡萄糖对s19细胞表达蛋白的影响。结果4个不同葡萄糖浓度实验组的细胞增殖速率及最大细胞密度均大于不加葡萄糖的对照组。两个较低葡萄糖浓度组的细胞凋亡率低于对照组，最高葡萄糖浓度组的细胞凋亡率则高于对照组。另外，加葡萄糖的8个实验组的蛋白表达量均高于不加葡萄糖的对照组，其中，3个较高葡萄糖浓度组显示有目的蛋白的降解条带。在另外5个没有出现降解条带的实验组中，以培养过程中间补加葡萄糖浓度至15mmol／L的实验组蛋白表达量最大。结论葡萄糖可促进s移细胞增殖，不同浓度葡萄糖对细胞凋亡的影响不同；适宜浓度的葡萄糖可提高sf9细胞表达完整外源蛋白的量。Objective To study effects of different glucose concentrations on growth characteristics of and protein expression in sf9 cells. Methods The densities of si9 cells cultured in Hyclone serum-free media supplemented with different concentrations of glucose were determined by trypan blue exclusion, and the effect of glucose concentration on cell proliferation and apoptosis rates was analyzed. The expression of human papillomavirus （HPV） type 18 L1 protein was determined by Western blot, and the effect of glucose concentra- tion on protein expression was studied. Results The cell proliferation rates and the largest cell densities in all test groups which were supplemented with glucose were higher than those in the control group which was not supplemented with glucose. The cell apoptosis rates in two test groups with lower concentrations of glucose were lower than that in the control group, while the apoptosis rate in the group with the highest concentration of glu- cose was higher than that in the control group. The expression of HPV18 L1 in all test groups were more than that in the control group. The degradation of HPV18 L1 was shown in three test groups with higher concentra- tions of glucose. The highest expression level of HPV18 L1 was observed in the group which was supplemented with glucose （ 15 mmoL/L） after a certain period of cell culture. Conclusions Glucose can improve cell proliferation. The cell apoptosis rates are different with glucose concentrations. Suitable concentration of glucose can elevate expression of HPV18 L1.

关 键 词：葡萄糖 昆虫细胞 增殖 蛋白表达 

分 类 号：R[医药卫生]