荞麦rBTI-2的表达及其对肿瘤细胞的生长抑制作用  

Expression of rBTI-2 of Buckwheat and Its Inhibit Proliferation Activity on Tumor Cells

作　　者：李玉英[1] 郭慧[1] 崔晓东[1] 王转花[1] 

机构地区：[1]山西大学生物技术研究所化学生物学与分子工程教育部重点实验室,太原030006

出　　处：《中国细胞生物学学报》2011年第7期759-765,共7页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(No.30671084;No.30870525);山西省自然科学基金(No.200701 1077)资助项目~~

摘　　要：设计一种适合基因工程开发的无标签重组荞麦胰蛋白酶抑制剂rBTI-2,并研究其对肿瘤细胞的生长抑制作用。构建原核表达载体pExSecI-BTI-2,诱导表达获得可溶性目的蛋白,经Resource^(TM) Q纯化后作用于HL-7702、HepG2、EC9706和QBC-939细胞,MTT检测rBTI-2对其生长的影响,并与前期获得的几种融合蛋白酶抑制剂进行功能比对。结果表明:质粒pEXSecI-BTI-2构建成功,SDS-PAGE分析表明分子量约为7.8 kDa。MTT检测表明rBTI-2对几种肿瘤细胞的生长有明显的抑制作用,而对正常细胞HL-7702作用很小。几种蛋白酶抑制剂对肿瘤细胞的生长均有不同程度的影响,其中rBTI-2对肿瘤细胞的生长抑制作用要大于融合蛋白酶抑制剂rBTI,这为深入研究BTI诱导肿瘤细胞凋亡的分子机制及其应用开发提供了重要基础和研究依据。To design a recombinant buckwheat trypsin inhibitor（rBTI-2） without label that suitable for developing genetic engineering and examine the effects of its inhibit proliferation activity on tumor cells, the pExSecI-BTI-2 was constructed. Soluble protein could acquired through inducing by IPTG. After purified through ResourceTM Q column, the possible effects of rBTI-2 on the proliferation of HL-7702, HepG2, EC9706 and QBC- 939 cell lines were investigated by MTT assays and compared the effect of several protease inhibitors on tumor cells. The results indicated that pExSecI-BTI-2 had been constructed. After purified and analyzed by SDS-PAGE, the approximate molecular weight was 7.8 kDa. MTT assays indicated that rBTI-2 could specifically inhibit the growth of HepG2, EC9706 and QBC-939, while it had showed less toxicity against HL-7702 cells. Several protease inhibitors could inhibit the proliferation of tumor cells on different level. Comparing with fusion protease inhibitors （rBTI）, the inhibit proliferation activity on tumor cells of rBTI-2 was slightly strong. It provides important foundation and evidence for the molecule mechanism of apoptosis and application of rBTI-2.

关 键 词：胰蛋白酶抑制剂 MTT检测 细胞增殖 

分 类 号：R285.5[医药卫生—中药学]