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Prognostic biomarkers and detection kits for patients with intrahepatic cholangiocarcinoma

A technology for detecting reagents and inner bile ducts, which is applied in the field of biomedicine, can solve the problems of complex evaluation calculation methods and limited consistency of evaluation results, and achieve the effects of stable prediction, high accuracy, and strong reliability

Active Publication Date: 2021-06-08
浙江高美生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the currently reported prognostic evaluation models usually include multiple factors such as tumor size, vascular invasion, tumor number, metastasis, and morphological characteristics. Patients need to undergo a large number of clinical medical tests, and the evaluation calculation methods are complicated, and the consistency of evaluation results is limited.
Moreover, there is currently no single biomarker or risk factor that can serve as a single predictor of ICC prognosis

Method used

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  • Prognostic biomarkers and detection kits for patients with intrahepatic cholangiocarcinoma
  • Prognostic biomarkers and detection kits for patients with intrahepatic cholangiocarcinoma
  • Prognostic biomarkers and detection kits for patients with intrahepatic cholangiocarcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] Example 1: Detection of the methylation level of the promoter region

[0096] First, genomic DNA was isolated using TiangenDP304 kit, and DNA degradation and contamination were monitored using agarose gel. DNA purity was measured with a spectrophotometer (IMPLEN, CA, USA). The DNA concentration was confirmed using the DNA Analysis Kit in the Qubit® 2.0 Flurometer, sonicated with a Covaris S220 to 200-300bp, followed by end repair and adenylation, and a total of 5.2 μg of genomic DNA fragments spiked with 26 ng λ DNA change. Ligate cytosine methylated barcodes to the sonicated DNA according to the kit manufacturer's instructions. The ligated DNA fragments were treated twice with bisulfite using the EZ DNA Methylation-Gold™ Kit (Zymo Research). Bisulfite-treated single-stranded DNA fragments were subjected to PCR amplification using the KAPA HiFi HotStartUracil + ReadyMix (2X) kit. Library concentration was quantified by Qubit 2.0 fluorometer (Life Technologies, CA, U...

Embodiment 2

[0100] Example 2 Obtaining the Methylation Score of the Promoter Region

[0101] Using the 24 promoter regions shown in Table 1, calculate PMS (promoter methylation score, promoter region methylation score). PMS is the sum of the product of the methylation level of each candidate promoter region and the corresponding coefficient. The prognosis assessment model for patients with intrahepatic cholangiocarcinoma has one or more optimal thresholds, and the optimal thresholds can correspond to the scores calculated by the model for assessing the prognosis of patients with intrahepatic cholangiocarcinoma to one of high PMS and low PMS. The group is either a group of high PMS, medium PMS and low PMS.

[0102] The detailed formula of PMS is shown in formula (1).

[0103] With 164 patients in the training cohort, the C-index of Overall Survival (OS) was 0.772 (95% CI: 0.731-0.813), and the optimal threshold of PMS was -11.20716.

[0104] With 170 patients in the validation cohort, t...

experiment example

[0111] Experimental example: PMS predictor, WCHSU predictor, JHUSM predictor, EHBSH predictor, AJCC TNM predictor, WCHSU-PMS factor effectiveness comparison

[0112] PMS predictors are as described in Example 1 and Example 2.

[0113] WCHSU: ascites status, tumor size, microvascular invasion status, lymph node metastasis status, tumor differentiation degree and CA19-9 carbohydrate antigen concentration, respectively.

[0114] JHUSM: age, tumor size, tumor number, lymph node status, microvascular invasion status, and liver cirrhosis (Y Wang et al, Prognostic Nomogram for Intrahepatic Cholangiocarcinoma After Partial Hepatectomy, J. Clin. Oncol. 31(2013) 1188-1195).

[0115] EHBSH: See O Hyder, H Marques et al, A Nomogram to Predict Long-term Survival After Resection for Intrahepatic Cholangiocarcinoma, JAMA Surg. 149(2014)432.

[0116] AJCC TNM: T staging mainly includes three main factors: tumor number, vascular invasion, and direct extrahepatic invasion; N staging is based o...

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Abstract

The present application discloses a prognostic biomarker and a detection kit for patients with intrahepatic cholangiocarcinoma. Using the set of polynucleotide sequences shown in SEQ ID NO.1‑SEQ ID NO.24 as biomarkers, methylation analysis was performed on these promoter regions, and an effective methylation scoring model was constructed to realize liver Accurate assessment of prognostic survival in patients with internal cholangiocarcinoma. This biomarker can be used as a single predictor of prognosis in patients with intrahepatic cholangiocarcinoma or as a predictor of survival in patients with intrahepatic cholangiocarcinoma based on multiple variables including gene promoter methylation scores.

Description

technical field [0001] The application relates to the field of biomedical technology, in particular to biomarkers and detection kits for the prognosis detection of patients with intrahepatic cholangiocarcinoma. Background technique [0002] Intrahepatic cholangiocarcinoma (ICC) is the second most common primary liver cancer, second only to hepatocellular carcinoma, with a high degree of malignancy and a very poor prognosis. Due to the insidious onset and high degree of malignancy of ICC, many ICC patients are not suitable for surgical treatment when they are first diagnosed. Nevertheless, surgical resection is still the most effective radical cure. However, the five-year survival rate of patients after surgical treatment is generally poor, only about 15%-45%. Even after radical resection, 57.9% to 73.4% of ICC patients still had recurrence, and 41.3% to 42.5% of ICC patients died of recurrence. [0003] For ICC patients who can be treated by surgical resection, accurately...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/117C12N15/11
CPCC12N15/117C12N2310/17C12Q1/6886C12Q2600/118C12Q2600/154C12Q2600/166
Inventor 孙德强
Owner 浙江高美生物科技有限公司
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