Kit for diagnosis and assessment of breast cancer, and detection and applications of methylated HOXA4/DPP6 gene
A HOXA4 and kit technology, applied in the field of biomedicine, can solve problems such as lack of research, and achieve the effects of good specificity, high accuracy and high precision.
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Embodiment 1
[0029] A kind of application of methylated HOXA4 / DPP6 gene, the accession numbers of described HOXA4 gene and DPP6 gene in the DNA sequence database are as shown in Table 1; The methylation of HOXA4 / DPP6 gene is CpG dinucleotide 5' The -terminal cytosine is converted to 5'-methylcytosine. Through the research of the present inventors, it is found that the methylation of HOXA4 / DPP6 gene in breast cancer tumors is greatly improved, wherein the methylation rate of HOXA4 gene in tumor tissue reaches 80.58%, and the methylation rate of DPP6 gene reaches 46.9%, which is higher than that in normal tissue. The methylation of HOXA4 / DPP6 gene was significantly increased. Therefore, the methylated HOXA4 / DPP6 gene can be used in the assessment, diagnosis, detection or screening of drugs in breast cancer.
[0030] Table 1: Accession numbers of the HOXA4 / DPP6 gene
[0031] gene symbol
Embodiment 2
[0033] A kit for diagnosing and evaluating breast cancer, said kit is a kit using methylated HOXA4 gene and / or methylated DPP6 gene as a biomarker, preferably simultaneously methylated HOXA4 / DPP6 Genes as biomarkers.
[0034] The kit includes specific primers designed for the biomarkers, and also includes TRIzol reagent (purchase manufacturer: Life Tech Inc, USA), 0.1M sodium citrate containing 10% ethanol, CT conversion agent (from EZ DNAMethylation TM Kit (ZYMO RESEARCH, Cat. No. D5002)), M-binding buffer, M-washing buffer, M-desulphonation buffer, bisulfite solution, Zymo Taq premix.
[0035] The above primers are to amplify the CpG of the 5'UTR of the methylated HOXA4 / DPP6 gene or the CpG of the exon 1 region to detect the methylation of the CpG, preferably including the amplification of the unmethylated HOXA4 / DPP6 gene Primers for the CpG of the 5'UTR or the CpG of the exon 1 region.
[0036] Specifically, the primers include a methylated HOXA4 gene primer and a methyla...
Embodiment 3
[0039] A kind of detection of methylation HOXA4 / DPP6 gene, described detection method is to take methylation HOXA4 / DPP6 gene as target, adopts illumina methylation chip (article number: WG-314-1003), and according to embodiment 2 The kit detects the degree of methylation of the HOXA4 / DPP6 gene. The specific method is referred to embodiment 4.
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