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Combined marker for early diagnosis of liver cancer and its application

A marker and liver cancer technology, applied in the field of biomedicine, can solve the problems of lack of effective therapeutic targets, difficulties in early diagnosis, high recurrence and metastasis rate, and achieve the effect of avoiding tissue biopsy, improving early screening sensitivity, and improving sensitivity

Active Publication Date: 2022-08-02
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current diagnosis and treatment of liver cancer still faces major problems: early diagnosis difficulties, high recurrence and metastasis rates, resistance to radiotherapy and chemotherapy, and lack of effective therapeutic targets, etc.
For nodules larger than 20mm, the diagnostic accuracy rate of ultrasonography is still 84%, but for intrahepatic nodules of 5-20mm, the sensitivity of ultrasound is only 33%.

Method used

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  • Combined marker for early diagnosis of liver cancer and its application
  • Combined marker for early diagnosis of liver cancer and its application
  • Combined marker for early diagnosis of liver cancer and its application

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0070] Example 1. Establishment of a method for detecting liver cancer

[0071] 1. Markers for liver cancer detection

[0072] The inventors of the present invention combined various databases and comprehensive clinical information, designed probes to enrich the methylation sites of related genes through large-scale screening, and then compared the cfDNA 3-6 months before the diagnosis of liver cancer through library construction and sequencing. Differences in methylation levels between samples and cfDNA samples from non-HCC patients to obtain markers for liver cancer detection. The markers include methylation levels of any 6 or more of the following 17 genes and / or mutations in at least one of the two genes TERT and CTNNB1:

[0073] AK055957, DAB2IP, GRASP, PPFIA1, PSD4, TBX15, MKL1, DCDC2, BCL2L11, SEMA4D, PROX1, FAM78A, PTPN7, AXIN1, SHH, SR, and CYP1B1.

[0074] The positions of these genes (reference genome GRCh38 / hg38) are shown in Table 1, wherein, 1) TBX15, MKL1, DCD...

Embodiment 2

[0125] Example 2. Using the 7 markers of the present invention to detect samples

[0126] The selected samples were 12 clinical liver cancer tissue samples and 12 healthy human leukocyte samples.

[0127] The inclusion criteria of 12 patients with clinical liver cancer were as follows: the diagnosis of liver cancer was confirmed by pathological examination of histological sections.

[0128] The inclusion criteria of 12 healthy people were: healthy non-HCC patients.

[0129] The seven markers are: MKL1, DCDC2, BCL2L11, CYP1B1, PTPN7, AXIN1 and SEMA4D gene methylation.

[0130] 1. The test sample is first passed through Qiagen's Genomic DNA was extracted using the Mini Kit kit according to the instructions in the instructions. The OD260 / 280 of DNA should be between 1.8-2.1 as determined by Nanodrop from ThermoFisher.

[0131] 2. Methylation detection was carried out to the extracted DNA, and according to the method of step 2 of Example 1, transformation, pre-amplification a...

Embodiment 3

[0137] Example 3. Using the 10 markers of the present invention to detect samples

[0138] The selected samples are the same as those in Example 2.

[0139] 10 markers: FAM78A, MKL1, DCDC2, BCL2L11, PTPN7, AXIN1, PROX1, AK055957, PPFIA1 and PSD4 gene methylation.

[0140] The detection steps are the same as those in Example 2. According to the methylation detection method in Step 2 of Example 1, the methylation of ten genes of FAM78A, MKL1, DCDC2, BCL2L11, PTPN7, AXIN1, PROX1, AK055957, PPFIA1 and PSD4 were detected. , and calculate the normalized value L of these ten genes respectively 目的基因 , the normalized value L for these ten genes 目的基因 Sum, get HCCscan, HCCscan=L FAM78A +L MKL1 +L DCDC2 +L BCL2L11 +L PTPN7 +L AXIN1 +L PROX1 +L AK055957 +L PPFIA1 +L PSD4 , the results of each sample are shown in Table 11.

[0141] Table 11. Test Results

[0142] sample name sample type HCCscan sample name sample type HCCscan T25 cancer tissue 105.4...

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Abstract

The invention discloses a combined marker for early diagnosis of liver cancer and its application. The combined markers for early diagnosis of liver cancer disclosed in the present invention are FAM78A, MKL1, DCDC2, SR, BCL2L11, CYP1B1, PTPN7, AXIN1, SEMA4D, PROX1, SHH, AK055957, DAB2IP, TBX15, GRASP, PPFIA1 and / or PSD4. Methylation, and mutation of TERT and / or CTNNB1. The present invention performs liver cancer detection by detecting the methylation level of the gene combination in the plasma cfDNA, and has high sensitivity and specificity; in the case of adding the mutation detection of the gene, by simultaneously detecting the methylation of the gene combination and the TERT gene , CTNNB1 gene mutation, effectively improve the sensitivity of early screening.

Description

technical field [0001] The invention relates to a combined marker for early diagnosis of liver cancer and its application in the field of biomedicine. Background technique [0002] Liver cancer is one of the malignant tumors with high incidence and great harm. However, the current diagnosis and treatment of liver cancer still faces major problems: difficulty in early diagnosis, high recurrence and metastasis rate, resistance to radiotherapy and chemotherapy, and lack of effective therapeutic targets. And for any cancer, early screening is always a critical step in the fight against cancer. [0003] The routine surveillance and screening indicators for the early diagnosis of liver cancer mainly include alpha-fetoprotein (AFP) and liver ultrasonography (US). Based on domestic and foreign reports and clinical practice, the diagnostic sensitivity of AFP is about 60%, the missed diagnosis rate is 40%, the specificity is about 80%, and the misdiagnosis rate is 20%. An important...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/154C12Q2600/156C12Q2600/166
Inventor 焦宇辰曲春枫钟夏宋欠欠王宇婷王沛郑乔松
Owner CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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