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Combined marker for early diagnosis of liver cancer and application of combined marker.

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

Active Publication Date: 2021-09-03
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 application of combined marker.
  • Combined marker for early diagnosis of liver cancer and application of combined marker.
  • Combined marker for early diagnosis of liver cancer and application of combined marker.

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Embodiment 1, the establishment of the method for liver cancer detection

[0071] 1. Markers for the detection of liver cancer

[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 by building a library and sequencing The difference in methylation level between samples and cfDNA samples of non-liver cancer patients was used to obtain markers for liver cancer detection. The markers include the degree of methylation of any 6 or more of the following 17 genes and / or the mutation of 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 T...

Embodiment 2

[0125] Example 2, using 7 markers of the present invention to detect samples

[0126] The selected samples were 12 clinical liver cancer tissue samples and 12 healthy white blood cell samples.

[0127] The inclusion criteria for 12 patients with clinical liver cancer were: liver cancer was diagnosed clinically by pathological examination of tissue sections.

[0128] The inclusion criteria of 12 healthy people were: healthy non-liver cancer 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 according to the operation in the instruction manual. The OD260 / 280 of DNA measured by ThermoFisher's Nanodrop should be between 1.8-2.1.

[0131] 2. The extracted DNA is subjected to methylation detection, according to the method of step 2 of Example 1, transformation, pre-amplification and final qPCR detection are carried out, and the re...

Embodiment 3

[0137] Example 3, using 10 markers of the present invention to detect samples

[0138] Selected sample is with embodiment 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 in Example 2, and the methylation of ten genes FAM78A, MKL1, DCDC2, BCL2L11, PTPN7, AXIN1, PROX1, AK055957, PPFIA1, and PSD4 are detected according to the methylation detection method in Step 2 of Example 1 , and calculate the normalized value L of these ten genes respectively 目的基因 , the normalized value L of these ten genes 目的基因 Sum to 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 105.40 WBC10 leukocyte 0.00 ...

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Abstract

The invention discloses a combined marker for early diagnosis of liver cancer and an application of the combined marker. The combined marker for early diagnosis of the liver cancer disclosed by the invention is methylation of FAM78A, MKL1, DCDC2, SR, BCL2L11, CYP1B1, PTPN7, AXIN1, SEMA4D, PROX1, SHH, AK055957, DAB2IP, TBX15, GRASP, PPFIA1 and / or PSD4, and mutation of TERT and / or CTNNB1. According to the invention, liver cancer detection is realized by detecting the methylation level of gene combination in plasma cfDNA, and the combined marker has high sensitivity and specificity; Under the condition of mutation detection after adding genes, the early screening sensitivity is effectively improved by detecting methylation of a gene combination and mutation of TERT genes and CTNNB1 genes at the same time.

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: early diagnosis difficulties, high recurrence and metastasis rates, resistance to radiotherapy and chemotherapy, and lack of effective therapeutic targets. And for any type of cancer, early screening is always a key step in fighting cancer. [0003] Routine monitoring and screening indicators for early diagnosis of liver cancer mainly include alpha-fetoprotein (AFP) and liver ultrasonography (US). Based on reports and clinical practice at home and abroad, the diagnostic sensitivity of AFP is about 60%, the misdiagnosis rate is 40%, the specificity is about 80%, and the misdiagnosis rate is 20%. An important reason for being a sc...

Claims

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Application Information

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Patent Type & Authority Applications(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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