Expression of biologically active human recombinant interferon alpha 2b in human breast cancer cell line Bcap-37
文献类型: 外文期刊
第一作者: Li, Hui
作者: Li, Hui;Shi, Zhendan;Li, Hui;Li, Xiangping;Liu, Qingyou;Shi, Deshun
作者机构:
关键词: Bcap-37;Biological active;IFN-inducible genes;Interferon alpha 2b
期刊名称:APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY ( 影响因子:2.926; 五年影响因子:2.685 )
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收录情况: SCI
摘要: Human interferon alpha 2b (IFNα-2b) is a pleiotropic cytokine used to treat various viral diseases and cancers. Conventionally, recombinant human IFNα-2b used in clinics was produced by prokaryotic expression system, which always lack of enough biological activity due to limitations on proper folding and post-translational modifications, so the eukaryotic expression system are becoming prevailing method for the production of recombinant proteins. In this study, human breast cancer cell Bcap-37 was firstly used as host for the expression of human IFNα-2b, with the expression vector pIRES2-IFN-EGFP, in which IFNα-2b gene is under the control of CMV promoter. The expression of recombinant IFNα-2b was detected by Western blot and ELISA. Results showed that the concentration of the secreted recombinant IFNα-2b in culture medium was 435.7 pg/mL/24 h. Biological activity of the recombinant IFNα-2b was assayed by detecting the expression of IFN-inducible genes, including MxA, OAS, PKR, and Caspase1 through QRT-PCR. Results demonstrated that recombinant IFNα-2b possess the biological activities. Compared to non-transgenic cells, the expression levels of the aforementioned four IFN-inducible genes were increased by 18.098-, 1.843-, 2.21-, and 3.066-folds, respectively. We got to a conclusion that the human breast cancer cell Bcap-37 could express bioactive recombinant IFNα-2b.
分类号: Q5
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