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Liver cancer tumor marker and application thereof

A tumor marker, liver cancer technology, applied to liver cancer tumor markers and their application fields, can solve the problems of lack of specific treatment methods, poor long-term prognosis of patients with hepatocellular carcinoma, and insidious onset, and improve the early diagnosis of liver cancer. The effect of horizontal, simple, fast and accurate detection

Inactive Publication Date: 2021-08-24
THE SECOND HOSPITAL OF NANJING +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, hepatocellular carcinoma is highly malignant and its onset is relatively insidious. When most cases are first diagnosed, there are already multiple metastasis in the liver, blood vessel and lymph node invasion, multiple metastasis in other organs, and even dyscrasia. Missed the best timing for surgery, and a small number of cases that meet the liver cancer resection criteria, even if the tumor has been successfully resected, still need to be assisted by multiple interventional treatments, radiofrequency ablation, molecular targeted drugs, traditional Chinese medicine and other comprehensive treatments
Long-term prognosis of patients with hepatocellular carcinoma remains poor even after multidisciplinary treatment
Moreover, liver cancer recurs early after surgery, and once it recurs, it progresses quickly, and there is a lack of specific treatment methods.

Method used

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  • Liver cancer tumor marker and application thereof
  • Liver cancer tumor marker and application thereof
  • Liver cancer tumor marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] This example illustrates the quantitative RT-PCR detection of the expression of the HOXB13 gene in liver cancer tissues.

[0033] 1. Tissue separation: The experimental tissue was derived from surgical patients with primary hepatocellular carcinoma. Once the surgically resected liver was isolated from the body, the lesion and surrounding paracancerous tissues 5 cm away were quickly excised and stored in liquid nitrogen (-80°C). The diagnosis of cancer and paracancer is based on pathological diagnosis.

[0034] 2. Extraction of ribonucleic acid (RNA): total RNA was extracted with a Trizol kit (Invitrogen), and the extraction method was referred to the instructions of the Trizol kit. When RNA is ready to be used, the RNA can be precipitated using the following method: add NaAc to 0.3M, centrifuge at 12,000×g for 5 minutes.

[0035] 3. Synthesis of cDNA:

[0036] (1) Add 4 μL of template RNA (0.5 μg / μL), 2 μL of 20 pmol / μL random primer, 5 μL of deionized water into the...

Embodiment 2

[0050] This example illustrates the use of quantitative RT-PCR to detect the expression of the HOXB13 gene in the overexpressed liver cancer cell line Hep3B.

[0051] Hepatocellular carcinoma Hep3B, Huh-7 cell lines and HL-7702 normal liver cell lines were selected, and after cell culture, WB expression was qualitatively and quantitatively performed. Wherein said cell line comes from Shanghai Cell Bank of Chinese Academy of Sciences.

[0052] According to the method in Example 1, RNA was extracted from different liver cancer cell lines, and cDNA was synthesized by reverse transcription. Using GAPDH as an internal control, and using the synthesized cDNA of different liver cancer cell lines as a template, the PCR reaction was carried out according to the method described in Example 1, and the agarose was gelled. PCR amplification products were detected by gel electrophoresis, the results are shown in figure 2 .

[0053] Depend on figure 2 It can be seen that HOXB13 gene pro...

Embodiment 3

[0055] The siRNA targeting HOXB13 was constructed, the specific sequence is shown in Table 2, and the liver cancer cell line was knocked down, and an empty vector (vector) was set as a negative control.

[0056] Table 2 HOXB13 siRNA sequences

[0057]

[0058] Two kinds of HOXB13 siRNA, negative control (NC) siRNA, empty lentiviral expression vector (Lenti-con), and HOXB13-expressing lentiviral expression plasmid (Lenti-HOXB3) were pressed into the transfection reagent using liposome (Lipofectamine 2000) respectively. Concentration of 50nmol / L was transfected into Huh-7 cells and Hep3B cells respectively, and cultured with DMEM containing 10% fetal bovine serum. According to the method described in Example 1, RNA was extracted from different liver cancer cell lines, and cDNA was synthesized by reverse transcription. Using GAPDH as an internal control, and using the synthesized cDNA of different liver cancer cell lines as a template, quantitative RT-PCR experiments were carr...

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Abstract

The invention relates to a liver cancer tumor marker and application thereof. The amino acid sequence of the liver cancer tumor marker is as shown in SEQ ID NO: 2, the nucleotide sequence is as shown in SEQ ID NO: 1, and the liver cancer tumor marker can be applied to the field of liver cancer diagnosis. The invention also relates to a detection primer pair of the liver cancer tumor marker, which has a nucleotide sequence as shown in SEQ ID NO: 3 and a nucleotide sequence as shown in SEQ ID NO: 4, and can be used as a liver cancer diagnostic reagent in the field of liver cancer diagnosis. The kit can simply, rapidly and accurately detect the liver cancer tumor marker, improves the early diagnosis level of liver cancer, and is suitable for large-scale popularization and application.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a liver cancer tumor marker and its application. Background technique [0002] The incidence of liver cancer has been increasing globally. By 2018, primary liver cancer has risen to become the fifth most common cancer in the world. Due to the poor prognosis of primary liver cancer, the mortality rate has increased, accounting for 8.2% of all cancer mortality. Currently, non-invasive liver stiffness testing, liver color Doppler ultrasound, serum alpha-fetoprotein, serum Golgi protein 73, including liver CT and MRI have become the main means for early diagnosis, treatment and follow-up of liver cancer. However, hepatocellular carcinoma is highly malignant and its onset is relatively insidious. When most cases are first diagnosed, there are already multiple metastasis in the liver, blood vessel and lymph node invasion, multiple metastasis in other organs, and even d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K14/82C12N15/12C12Q1/6886C12N15/11G01N33/574
CPCC07K14/4702C07K14/82C12Q1/6886C12Q2600/158G01N33/57438G01N33/57484G01N2333/4703
Inventor 左凌云陈雨欣谭婷婷王华利涂宏飞
Owner THE SECOND HOSPITAL OF NANJING
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