NF-ª—B inhibitor, preparing process thereof and application as antineoplastic medicine

Abstract

The invention relates to a NF-kappaB inhibitor, process for preparation, and use as antineoplastic medicine, wherein extracorporal pointing mutation method is applied twice for performing orienting mutagenesis to the codon of the No.32 and No.36 bit encoding serine to construct IkappaBalphaM super inhibitor, which is than sub-cloned from pGEM-T carrier into pAdTrack-CMV shuttle vector, then it is expressed in pronucleus and eukaryotic cells (including 293.HepG2, HeLa, HUVEC, MAD-435S cells, etc.), thus obtaining the protein (expressed NF-kappaBsuperinhibitor IkappaBalphaM).

Description

Technical field: [0001] The invention belongs to the technical field of genetic engineering. Background technique: [0002] At present, the core problem of tumor treatment is to solve the apoptosis-inducing effect of anti-cancer drugs on tumor cells. Recent studies have found that the abnormal activation of nuclear transcription factor NF-κB plays a key role in the anti-apoptotic mechanism of tumor cells. Under normal circumstances, NF-κB binds to its inhibitor IκBα and does not exhibit biological activity. When cells are stimulated by external factors such as TNF-α, a series of enzymatic cascade reactions will cause IκBα to be phosphorylated at Ser32 and Ser36. Ubiquitination and enzymatic hydrolysis rapidly, and finally activate NF-κB and combine with specific target genes in the nucleus to start the expression of stress proteins to resist TNF-α and other effects. This discovery reveals the root cause of tumor cell drug resistance. Therefore, how to inhibit the function ...

AI Technical Summary

Problems solved by technology

[0004] The construction methods of NF-κB inhibitors (IκBαM super-inhibitors) have been reported abroad, and the methods are different, and the codons encoding serine at the 32nd and 36th positions were directional induced twice by the in vitro site-directed mutagenesis method. The method of preparing super-inhibitor IκBαM and then using specific primers for PCR amplification has not yet been reported at home and abroad. Human hepatoma cell line HePG 2 The influence of cell nucleus morphology, the study of anti-tumor activity in vivo, etc., have not been reported so far

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