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Molecular marker and kit for diagnosing cancer

A kit and cancer technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems that patients cannot accept surgery

Pending Publication Date: 2022-03-08
THE FIRST MEDICAL CENT CHINESE PLA GENERAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The only cure for PDAC is surgical resection (Masiak-Segit W, Rawicz-Pruszyński K. Surgical treatment of pancreatic cancer. Pol Przegl Chir. 2018 Apr 30; 90(2):45-53.); however, due to locally advanced or Metastatic disease, approximately 80% of patients are inoperable

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  • Molecular marker and kit for diagnosing cancer
  • Molecular marker and kit for diagnosing cancer
  • Molecular marker and kit for diagnosing cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] 1. Template preparation (extraction of genomic DNA and sulfuration modification process)

[0045] Preparation of DNA: Pancreatic cancer, breast cancer, esophageal cancer specimens and normal tissue specimens mentioned above were obtained. In this example, 5 cases of pancreatic cancer (PC1-PC5), 5 cases of breast cancer (BC1-BC5), 7 cases of esophageal cancer (EC1-EC7), 2 cases of normal pancreas (PN1-PN2), and 2 cases of normal breast cancer (BN1-BN2) and 2 cases of normal esophagus (EN1-EN2), the genomic DNA was extracted by phenol-chloroform extraction, and its content and purity were determined by measuring the absorbance (A) value with a UV spectrophotometer.

[0046] Sulfite modification: refer to Herman (J.G.Herman, J.R.Graff, S.Myohanen, B.D.Nelkin and S.B.Baylin, Methylation-specific PCR: a novel PCR assay formethylationstatus of CpG islands, Proc.Natl.Acad.Sci.USA93(1996 ), 9821–9826.) and other methods to improve. Take the genomic DNA prepared above, after d...

Embodiment 2

[0067] Example 2 Clinical Specimen Detection

[0068] Take 204 cases of pancreatic cancer clinical samples, 12 cases of breast cancer samples, and 67 cases of esophageal cancer samples for MS-PCR amplification. Template preparation, PCR amplification system and conditions, and detection of amplification products are all the same as those in Example 1. , the test results are shown in the table below:

[0069]

Embodiment 3

[0070] Embodiment 3 sensitivity experiment

[0071] The DNA of pancreatic cancer cell line SW1990 (100% methylation in the promoter region of BEND4 gene) and the DNA of normal lymphocytes (100% non-methylation in the promoter region of BEND4 gene) were mixed in proportion to carry out sulfuration modification (the method is the same as that in the implementation Example 1), then perform MS-PCR. PCR products were subjected to 2% agarose gel electrophoresis, measured by ultraviolet transmission analyzer and photographed.

[0072] Grouping: Group 1: 100% SW1990 cell DNA+0% normal cell DNA

[0073] Group 2: 50% SW1990 cell DNA+50% normal cell DNA

[0074] Group 3: 5% SW1990 cell DNA + 95% normal cell DNA

[0075] Group 4: 1% SW1990 cell DNA + 99% normal cell DNA

[0076] Group 5: 0.1% SW1990 cell DNA + 99.9% normal cell DNA

[0077] Group 6: 0% SW1990 cell DNA + 100% normal cell DNA

[0078] See the result figure 2 : Only one pancreatic cancer cell in 1000 normal cells can...

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Abstract

The invention discloses a molecular marker and a kit for cancer diagnosis, and particularly relates to application of BEND4 gene promoter region DNA methylation as a molecular marker in preparation of cancer diagnosis products. According to the application disclosed by the invention, frequent high methylation of a BEND4 promoter region in pancreatic cancer, breast cancer and esophageal cancer is found through an MSP method for the first time, BEND4 participates in an NHEJ pathway of DNA damage repair, and BEND4 methylation is a synergistic lethal marker for killing tumor cells by combining an ATM inhibitor with chemotherapeutic drugs such as cis-platinum and the like. The invention also provides a primer and a kit for detecting the methylation state of the promoter region of the BEND4 gene. The application of the primer and the kit to detect the hypermethylation state of the BEND4 gene promoter region can be used as a powerful means for diagnosis, curative effect observation, prognosis judgment, minimal residual disease detection and the like of pancreatic cancer, breast cancer and esophageal cancer, and the primer and the kit are simple and convenient to operate and good in stability and have profound clinical significance and generalization performance.

Description

technical field [0001] The present invention relates to the field of biotechnology. More specifically, it relates to a molecular marker and a kit for diagnosing cancer. Background technique [0002] Pancreatic cancer (PC) is one of the most refractory cancers and is considered to have the worst prognosis among digestive system tumors. Despite the use of different surgical treatment options and adjuvant therapy and neoadjuvant chemotherapy to treat PC, the overall 5-year survival rate still does not exceed 8% (Bray F, Ferlay J. Global cancer statistics 2018: GLOBO CAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018 Nov;68(6):394-424.; Siegel RL, Miller KD. CA Cancer J Clin. 2017 Jan; 67(1):7-30.). The high mortality rate of PC patients is mainly attributed to its lack of typical symptoms and aggressive local invasion in its early stage. PDAC (pancreatic ductal adenocarcinoma, PDAC) is by far the most common type, accou...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/118C12Q2600/154C12Q2600/166
Inventor 郭明洲张美英吕红慧苏小茉姚远新于咏
Owner THE FIRST MEDICAL CENT CHINESE PLA GENERAL HOSPITAL