Sensitive diagnosis and relative treatment for cancers through 20 tumor specific related genes and products
A tumor and cancer technology, applied in gene therapy, microbial-based methods, anti-tumor drugs, etc., can solve problems such as blindness of treatment effect, high cost, and lack of patients
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Embodiment 1
[0139] Example 1. Identification of target genes of gene promoter DNA methylation in human normal tissues:
[0140] DNA methylation of gene promoters in the human genome is one of the important steps in the regulation of gene expression in the process of tissue differentiation. In the early stage of tumor onset, many important genes related to tissue differentiation are deregulated by methylation regulation of tumor cells. The original silent state becomes active expression, human AFP (hepatic alpha-fetoprotein) is a good example. This experiment aims to obtain those gene promoters in the human genome that are highly methylated in normal conditions, but in tumor cells It may be the demethylation state. This information provides the basis for searching for gene demethylation in tumor cells. This experiment uses the CHIP_DSL technology system of AVIVA SYSTEMS BIOLOGY (ASB) company in the United States. This technology system has been published in PNAS in 2006 Published in (March...
Embodiment 2
[0150] Example 2. An experiment to identify tumor-specifically expressed genes.
[0151] Because the present invention aims to obtain tumor-specific target genes, the second step of the experiment focuses on finding genes that are basically not expressed in normal tissues but highly expressed in tumor tissues.
[0152] According to the introduction in Experimental Protocol 1, many early embryonic stage-specific genes are abnormally highly expressed in adult tumor tissue cells due to the dysregulation of methylation regulation in many tumor tissue cells. Such genes are often turned off in normal tissues Good but not expressed at all. According to this theory, the second step of the experiment focused on the identification of specific expression genes in the early embryo. Since the germ cells in the human body are closest to the early embryo state, the mRNA of various human tissues and human germ cells was used. The results of the chip assay library for specific expression were ...
Embodiment 3
[0154] Example 3, production of tumor-specific expression gene protein target antibody:
[0155] In order to further prove that the 172 target genes in tumor cells do have related protein expression in different tumor cells, we further selected 20 target gene proteins for rabbit polyclonal antibody production.( image 3 There are 20 kinds of target gene names and NCBI gene bank gene sequence numbers). Among them, 14 target proteins obtain their specific polypeptide antigens by peptide synthesis, and each target protein has two specific polypeptide antigens. The other 6 target proteins are all Long gene recombination to obtain protein antigen for production of target protein specific antibody.
[0156] 3A: Peptide synthesis of target protein:
[0157] The peptide synthesis of 14 target proteins was carried out routinely in the operation instruction manual provided by ACT Company in the Apex Polypeptide Synthesis Instrument of ACT Company: Wang resin was used as a solid phase c...
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