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Expression of rBTI-2 of Buckwheat and Its Inhibit Proliferation Activity on Tumor Cells
Li Yuying*, Guo Hui, Cui Xiaodong, Wang Zhuanhua
Key Laboratory for Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology,Shanxi University, Taiyuan 030006, China
Abstract: 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 Resource™ 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.
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