Neural protein NSPc1 having influence on nerve tumor cell differentiation by tretinoin signal pathway

Abstract

It is found out in the invention that the poly-comb protein NSPc1 of the neural system can highly express in the tumor cell lines from a variety source of neural system using the Western blot technique, it is also found out that the expression level of NSPc1 in human gliocytoma sample is far higher than that in the normal tissue around the cancer and it goes up as the grading increase of the neurogliocytoma pathology using the Real-time PCR technique. NSPc1 is found to have transcription inhibition capacity using the fluorescence detection of reporting gene technology, and p21Waf1 / Cip1 is found to be the target controlled gene of NSPc1. Promoter sequence mutation experiments and chromatin co-immunoprecipitation experiments of P21Waf1 / Cip1 showed that the inhibition effect of NSPc1 on the transcription of p21Waf1 / Cip1 depends on retinoic acid response elements (RARE). Overexpressions of retinoic acid receptor RXRalphacan eliminate the inhibition activity of NSPc1 on p21Waf1 / Cip1 promoter. These results indicate that NSPc1 highly expressing in nerve tumor inhibits the transcription of p21Waf1 / Cip1 through interfering the controlling access of the retinoic acid thus affects the differentiation of the nervous system tumor. Therefore, not only can NSPc1 be used as candidate markers for grouping diagnostic of nervous system malignant tumor type, but also can serve as a target of gene therapy. Meanwhile, in the invention, building the plasmid for prokaryotic expressing of full-length protein of NSPc1, transforming Escherichia coli to express the corresponding protein; After purifying the protein it can be used for the preparation of high-quality antibodies, as well as applied to the nervous system tumor diagnosis.

Description

technical field

[0001] The invention belongs to the field of gene function, antibody preparation and clinical diagnosis. Specifically, the present invention relates to a full-length recombinant genetic engineering and protein purification process of a neural tumor cell differentiation-related gene NSPc1, involves the use of purified protein to prepare highly specific NSPc1 antibodies, and involves the use of Western Blot technology and Real-time PCR technology to detect The expression level of NSPc1 in tumor cell lines derived from nervous system and human glioblastoma specimens, involving the confirmation of NSPc1 downstream target gene p21Waf1 / Cip and the discovery of NSPc1 regulatory sequence retinoic acid receptor response element (RARE) and related regulation Confirmation of the mechanism, involving recombinant plasmids containing the NSPc1 gene and recombinant strains expressing the protein and their application in the diagnosis of glioblastoma, involving the NSPc1 prote...

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