Primer and kit for detecting serum exosome RNF144A-AS1 and application of primer and kit in diagnosis and treatment of liver metastasis of gastric cancer

A technology of RNF144A-AS1, exosomes, applied in DNA/RNA fragments, recombinant DNA technology, determination/inspection of microorganisms, etc.

Inactive Publication Date: 2020-11-13
ZHENJIANG NO 1 PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It has been reported that CD44v6, OPN, YB-1, MAGE-A10 and other molecules may be used as...

Method used

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  • Primer and kit for detecting serum exosome RNF144A-AS1 and application of primer and kit in diagnosis and treatment of liver metastasis of gastric cancer
  • Primer and kit for detecting serum exosome RNF144A-AS1 and application of primer and kit in diagnosis and treatment of liver metastasis of gastric cancer
  • Primer and kit for detecting serum exosome RNF144A-AS1 and application of primer and kit in diagnosis and treatment of liver metastasis of gastric cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Detection of RNF144A-AS1 in serum exosomes

[0032] 1. Obtain the serum sample to be tested:

[0033] Serum samples from 120 cases of gastric cancer and 120 normal persons.

[0034] The samples are the serum samples of the patients with gastric cancer and normal people with coagulant added.

[0035] 2. Extract exosomes from serum samples to be tested and verify

[0036] Extract exosomes from gastric serum according to the instructions of the exosome extraction kit. The steps are as follows: (1) Whole blood is centrifuged at 3000×g for 15 min (centrifugal radius 10 cm) to remove cells or cell debris. (2) Put the upper layer liquid into a centrifuge tube, add 63 μl ExoQuick reagent to 250 μl serum, put it at 4°C for 30 minutes; (3) Centrifuge the mixture at 1500×g (centrifugal radius 10cm) for 30 minutes at 4°C; ( 4) Aspirate all the supernatant, centrifuge at 1500×g (centrifugal radius 10cm) for 5 min at 4°C, and aspirate the entire supernatant; (5) Dissolve...

Embodiment 2

[0066] Example 2 Clinical significance of serum exosome RNF144A-AS1

[0067] The correlation between serum exosomal RNF144A-AS1 expression and clinical parameters such as gender, age, tumor diameter, clinical stage, tissue differentiation, lymph node metastasis, and liver metastasis of gastric cancer was further analyzed. See Table 4 and Table 5. It was found that the expression level of serum exosomal RNF144A-AS1 was significantly correlated with the depth of invasion, lymph node metastasis, liver metastasis, and clinical stage of patients (P values ​​were 0.014, 0.014, 0.003, and 0.001), while it was significantly correlated with Gender, age, tumor location, tumor diameter, tissue differentiation, Helicobacter pylori infection and CEA level were not significantly correlated (P values ​​were 0.577, 0.852, 0.545, 0.201, 0.548, 0.143 and 0.583, respectively). The results of multivariate analysis suggested that the high expression of serum exosomal RNF144A-AS1 was one of the ind...

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Abstract

The invention relates to primers and a kit for detecting serum exosome RNF144A-AS1 and application of the primer and the kit in diagnosis and treatment of liver metastasis of gastric cancer. The kit comprises an RNA extraction reagent of a serum lncRNA sample, a primer, a genome-removed DNA reaction system, a reverse transcription reaction system and a qPCR operation reaction system. The inventiondiscloses an expression condition of lncRNA in serum exosomes of gastric cancer patients and normal people, and RNF144A-AS1 is detected by using fluorescent quantitative PCR. The serum exosome RNF144A-AS1 can be used for early diagnosis of liver metastasis of gastric cancer, judging disease progression and prognosis conditions and evaluating the treatment effect of liver metastasis of gastric cancer.

Description

technical field [0001] The invention relates to primers and a kit for detecting serum exosome RNF144A-AS1 and its application in the diagnosis and treatment of gastric cancer liver metastasis, belonging to the technical field of molecular diagnosis. Background technique [0002] Gastric cancer is prone to liver metastasis, which is inseparable from the biological characteristics of the liver. The liver is the largest glandular organ in the human body, and is supplied by both the hepatic portal vein and the hepatic artery. Among them, the hepatic portal vein accounts for 80%, and the hepatic artery accounts for 20%. Hepatic capillaries are porous sinusoidal structures with strong permeability, and hepatic sinusoidal ducts lack a subcutaneous basement membrane. Compared with other organs such as brain, lung, and bone, tumor cells are more likely to extravasate in the liver. Therefore, the liver is the most important organ for gastrointestinal tumor metastasis. [0003] Gastr...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/113
CPCC12Q1/6886C12Q2600/158C12Q2600/178C12Q2600/118
Inventor 范钰何蓉郎亚昆满昌峰周永静龚丹丹
Owner ZHENJIANG NO 1 PEOPLES HOSPITAL
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