Cancer screening method

A screening, cancer technology, applied in the fields of biochemical equipment and methods, microbial determination/inspection, etc.

Active Publication Date: 2010-10-20
NAT DEFENSE MEDICAL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] It can be seen that there are still many deficiencies in the above-mentioned existin...

Method used

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Experimental program
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Embodiment 1

[0039] Embodiment one material and method

[0040] 1. Materials

[0041] The test materials included a series of complete samples of cervical lesions, including: squamous cell carcinoma (SCC, n=20), adenocarcinoma (adenocarcinoma, AC, n=20), and normal cervical samples (n=10 ). All cervical samples, ovarian samples, and colorectal cancer samples were taken from the Taipei Tri-Service General Hospital. The genomic DNA (genomic DNA) of each sample was extracted with the QIAamp DNA kit (QIAGEN) and used to compare and analyze DNA methylation in the whole genome situation. In addition, the quality of genomic DNA was detected by Bioanalyzer (Agilent) before analysis. In this embodiment, 10 μg of fragmented DNA was used for MeDIP (Methyl DNA IP).

[0042] 2. DNA methylation analysis by MeDIP and CpG island-Plus-Promoter arrays (CpGisland-Plus-Promoter arrays)

[0043] First, with Bioruptor TM UCS-200 (Diagenode) fragments genomic DNA to 300-1,000 bp. Then add 30 μl polyclona...

Embodiment 2

[0055] Example 2 Screening of cervical cancer methylation target genes

[0056] After screening by CpG island-Plus-Promoter arrays (CpG island-Plus-Promoter arrays), four target genes were screened out that may have hypermethylation in cervical cancer cells, namely DBC1 (SEQ ID No: 1 ), PDE8B (SEQ ID No: 2), PTPRR (SEQ ID No: 3) and ZNF582 (SEQ ID No: 4), the details of which are shown in Table 3; it can be seen from Table 3 that these four genes have been Apart from bladder cancer, few studies have shown a link between these genes and cervical cancer.

[0057] Table 3 Details of methylated genes in cervical cancer cells screened by CpG island-Plus-Promoter array

[0058]

[0059]

Embodiment 3

[0060] Example Trisulfite sequencing (BS) analysis of methylation status of target genes in cervical lesion samples

[0061] Target gene: PTPRR

[0062] Test sample group:

[0063] 1. HeLa cervical cancer cell line (HeLa_0);

[0064] 2. HeLa cervical cancer cell line (HeLa_10D) treated with 10 μM DNA methyltransferase inhibitor 5'-aza-2'-deoxycytidine (AZC);

[0065] 3. Simultaneously treat the HeLa cervical cancer cell line ( HeLa_DT);

[0066] 4. Cervical adenocarcinoma sample (AC);

[0067] 5. Cervical squamous cell carcinoma samples (SCC);

[0068] 6. Control group (normal): Normal cervical blood DNA was used as the control group without methylation.

[0069] After the aforementioned test samples were modified with sulfite, followed by sulfite sequencing (BS) to analyze whether the target gene (PTPRR) had hypermethylation (hypermethylation) phenomenon in each test sample, the results are shown in Figure 1, Black represents methylated regions and white represents u...

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Abstract

The invention relates to a cancer screening method which comprises the following steps that: (1) providing a tested body; (2) testing the CpG sequence methylation state of at least one target gene in genome DNA of the tested body, and the target gene comprises PTPRR, ZNF582, PDE8B and DBC1; (3) judging whether the tested body has cancer or lesions before cancer according to that whether the methylation state of the at least one target gene exists; and the methylation state detection method is methylation-specific PCR, MSP, quantitative methylation-specific PCR, QMSP, bisulfite sequencing (BS), microarrays, massspectrometer, denaturing high-performance liquidchromatography (DHPLC) and the like.

Description

technical field [0001] The invention relates to a cancer screening method, in particular to a cancer screening method using methylated DNA as a biomarker. Background technique [0002] Cervical cancer is one of the main causes of death for women in the world and in Taiwan. According to the statistics of the World Health Organization (WHO) in 2002, cervical cancer is the second leading cause of cancer death for women in the world, second only to breast cancer; regular cervical cancer screening Cervical cancer screening is the best way to prevent cervical cancer. There are two main methods of cervical cancer screening, one is the most common pap smear (Pap smear), and the other is human papillomavirus (HPV) test. testing); Cervical smear is to take out the secretions of the cervix, and use a microscope to observe whether there are cancerous lesions in the exfoliated epithelial cells, so as to detect cervical cancer at an early stage; Reaction (polymerase chain reaction, RT-PC...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 赖鸿政
Owner NAT DEFENSE MEDICAL CENT
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