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Serum miRNA marker and application of serum miRNA marker in early diagnosis for pancreatic cancer induced by pancreatitis

A marker and serum technology, applied in the field of molecular biology, can solve the problems of poor stability and repeatability of chip technology information quality, high detection cost, and high miRNA purity requirements.

Inactive Publication Date: 2019-06-18
THE CENT HOSPITAL OF WUHAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Chinese authorized patent CN1028766B relates to a serum / plasma miRNA marker related to pancreatic cancer and its application. The marker is a combination of miR-451 and miR-409-3p. The application of the relevant kit requires the corresponding 4 kinds of specific primers, the detection cost is high
On the other hand, the published Chinese patent CN108103198A discloses a plasma miRNA marker and its application related to the auxiliary diagnosis of pancreatic cancer. The related markers are miR122-5p, miR-125b-5p, miR-192-5p, miR-193b3p One or more of miR-221-3p and miR-27b-3p, one or more specific primers required are provided in the kit, through Exiqon miRNAqPCR panel chip and qRT-PCR-based absolute quantification method, However, microarray detection has high requirements on the purity of miRNA, and the stability and repeatability of microarray technology information quality are poor

Method used

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  • Serum miRNA marker and application of serum miRNA marker in early diagnosis for pancreatic cancer induced by pancreatitis
  • Serum miRNA marker and application of serum miRNA marker in early diagnosis for pancreatic cancer induced by pancreatitis
  • Serum miRNA marker and application of serum miRNA marker in early diagnosis for pancreatic cancer induced by pancreatitis

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Experimental program
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Effect test

Embodiment 1

[0032] Embodiment 1, embodiment 2, embodiment column 3 and embodiment 4 provide a kind of miRNA detection kit, comprise following components:

[0033]

[0034]

[0035] The composition of the above kit can be used for 100 times of fluorescent quantitative PCR reactions and stored at -20°C. The sources of the above reagents are provided by the above reagent companies.

[0036] The present invention provides the application of the miRNA detection kit in the early diagnosis of pancreatic cancer mediated by pancreatitis, and the specific implementation method is as follows:

[0037] Example 1: Detection of expression levels of miR-301a in pancreatitis, pancreatic cancer patient plasma, pancreatic cancer tissues and cell lines using the above kit

[0038] 1. Specimen collection

[0039] Plasma samples were collected from 70 non-small cell pancreatic cancer patients with a history of pancreatitis who were clinically confirmed and had not received any treatment. Plasma sample...

Embodiment 2、 Embodiment 3、 Embodiment 4

[0078] Example 2, Example 3, Example 4: Using the above kit to detect the expression level of miR-301a in the plasma of patients with pancreatic cancer

[0079] A kit for the early diagnosis of inflammation-mediated pancreatic cancer provided in Example 2, Example 3, and Example 4 differs from Example 1 in that 10 μM miRNA in the reverse transcription (RT-PCR) reaction system reverses the The volume ratios of primers, 10 μM U6 reverse transcription primer, nuclease-free water, 5× reverse transcription buffer and enzyme preparation are different, see Table 2:

[0080] Reagent volume in table 2 reverse transcription reaction system

[0081]

[0082] Quantitative PCR detection of plasma miR-301a expression in 63 patients with pancreatic cancer, see Figure 6 The volume ratio of the 10 μM miRNA reverse transcription primer, 10 μM U6 reverse transcription primer, nuclease-free water, 5× reverse transcription buffer, and enzyme preparation used in Example 1 was 1:1:16.75:5:1.25,...

Embodiment 5

[0083] Example 5: Correlation analysis of plasma miR-301a level in patients with pancreatic cancer and pancreatitis-related factors IL-6, TNF-α

[0084] 1. Analysis method: SPSS 19.0 software was used to conduct Pearson correlation test to analyze the correlation between plasma miR-301a level and pancreatitis markers.

[0085] 2. Experimental results: analysis of the correlation between plasma miR-301a level and common pancreatitis markers IL-6 and TNF-α, see Figure 7 , Figure 8 , the results of Pearson correlation test analysis showed that the correlation coefficients R of plasma miR-301a, IL-6 and TNF-α in patients with pancreatic cancer were 0.35 and 0.65, respectively, P<0.01, indicating that the plasma miR-301a level and IL-6 -6 and TNF-α have a certain correlation, further indicating that the level of plasma miR-301a has a certain correlation with the well-known carcinoembryonic antigen CEA and pancreatic cancer marker CA199, circulating miR-301a is expected to play a...

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Abstract

The invention discloses a serum marker miR-301a with the sequence of SEQ ID NO. 1 and application of a kit prepared by adopting the miR-301a marker as a molecular marker in early diagnosis for pancreatic cancer induced by pancreatitis and curative effect monitoring. The kit prepared by adopting the miR-301a marker can conveniently and quickly detect the expression level of miRNA, it is not necessary to customize special primer sequences additionally, and by detecting the blood plasma, pancreatic cancer tissue and expression level of the miR-301a in a pancreatic cancer cell line of a patient suffering from pancreatitis and pancreatic cancer, the Pearson correlation test is utilized for analyzing the relationship between the miR-301a and a known traditional marker of pancreatitis and comparing the miR-301a with the known traditional marker of pancreatitis; through a receiver operating characteristic curve (ROC), the identification capacity of the miR-301a and joint detection by the miR-301a and CA199 in pancreatic cancer diagnosis and pancreatic cancer staging is evaluated, and the potential value of the miR-301a marker of the blood plasma in early diagnosis for pancreatic cancer induced by pancreatitis is investigated to provide a new theory and a new test basis for early diagnosis and targeted therapy of pancreatic cancer from the miRNA level.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to the application of a serum marker miR-301a in the early diagnosis and curative effect monitoring of pancreatic cancer mediated by pancreatitis. Background technique [0002] Pancreatic cancer has a high degree of malignancy and is one of the tumors with the worst prognosis in China for digestive system tumors. Statistics released by the World Health Organization show that the incidence of pancreatic cancer in the world ranks 13th among malignant tumors, but its fatality rate is the 13th. 4 bit. In my country, pancreatic cancer is the seventh leading cancer-related death. Early pancreatic cancer usually has no obvious clinical symptoms and signs, and has the characteristics of low early diagnosis rate, poor curative effect, and poor prognosis. Surgery is currently the main treatment for pancreatic cancer. However, the surgical resection rate is only 10% to 20...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/6886
Inventor 卢忠心胡绘陈卫群吴唐维李晓怡刘水逸
Owner THE CENT HOSPITAL OF WUHAN
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