Application of pattern recognition receptor RIG-I in preparing kit for tumor diagnosis or prognosis evaluation

A pattern recognition receptor, tumor diagnosis technology, applied in the field of biotechnology and medical diagnosis

Inactive Publication Date: 2014-04-02
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0017] At present, there is no research report on the correlation between the pattern recognition receptor RIG-I and tumors at home and abroad.

Method used

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  • Application of pattern recognition receptor RIG-I in preparing kit for tumor diagnosis or prognosis evaluation
  • Application of pattern recognition receptor RIG-I in preparing kit for tumor diagnosis or prognosis evaluation
  • Application of pattern recognition receptor RIG-I in preparing kit for tumor diagnosis or prognosis evaluation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Embodiment 1: the preparation of detection kit

[0083] The detection kit is prepared according to the following composition, and the kit is suitable for detecting the expression of RIG-I in a biological sample with real-time quantitative reverse transcription PCR (qRT-PCR) method:

[0084] (a) a container with reverse transcriptase (200U / μl);

[0085] (b) a container containing an RNase inhibitor (40 U / μl);

[0086] (c) Equipped with reverse transcription 5× buffer (75mM KCl, 500mM Tris-Cl, pH8.3, 25°C, 3mMMgCl 2 , 10mM DTT) container;

[0087] (d) A container containing target gene reverse transcription primers and PCR upstream primers and downstream primers (10 μM): RIG-I reverse transcription primers are: Oligo (dT);

[0088] Quantitative PCR primers:

[0089] 5'-TGT GCT CCT ACA GGT TGT GGA-3' (upstream, SEQ ID NO: 1) and

[0090] 5'-CAC TGG GAT CTG ATT CGC AAA A-3' (downstream, SEQ ID NO: 2);

[0091] (e) A container containing internal reference reverse tran...

Embodiment 2

[0100] Example 2: Relationship between RIG-I expression level and HCC

[0101] Adopt real-time quantitative reverse transcription PCR (qRT-PCR) method, use the detection kit among the embodiment 1 to collect in 1999 to 2006 the RIG-1 mRNA level in the HCC tissue of two groups of 292 routine HCC patients altogether and the paracancerous tissue (Group 1 tissue sections were from Sun Yat-Sen University, Guangzhou, and Group 2 tissue sections were from the Affiliated Hospital of Guangxi Medical University).

[0102] Total tissue RNA was extracted using TRIzol (Invitrogen). qRT-PCR was completed on a LightCycler 1.5 (Roche Company) real-time quantitative PCR instrument using the test kit of Example 1.

[0103] Relative quantification of RIG-I using 2 -ΔΔCt (β-actin as internal reference) (Livak, KJ. ​​et al., Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method. Methods. 2001; 25: 402-408).

[0104] The results of qRT-PCR analysis show...

Embodiment 3

[0122] Sample: a tissue section of a liver cancer patient's tumor, and the relative expression level of RIG-I in the liver cancer tissue was detected using the kit of Example 1. After calculation, the expression level of RIG-I in the cancer tissue was -9.11, which was 30% lower than that of the normal control.

[0123] According to postoperative follow-up, the patient died 6 months after the operation, and the disease-free survival period was only 3 months.

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Abstract

The invention relates to the fields of biotechnology and medical diagnosis. The research finds that the molecular expression of a pattern recognition receptor RIG-I in the tumor tissue of a tumor patient is obviously decreased and the correlation exists between the decreasing amplitude and the survival time of the tumor patient; moreover, the low expression of the molecules of the pattern recognition receptor RIG-I is significantly correlated with the prognosis difference of the patient. The invention provides an application of the pattern recognition receptor RIG-I in preparation of a kit for tumor diagnosis or prognosis evaluation, and the corresponding detection kit.

Description

technical field [0001] The invention relates to the fields of biotechnology and medical diagnosis, in particular to the application of pattern recognition receptor RIG-I in the preparation of tumor diagnosis or prognosis evaluation kits. Background technique [0002] In the process of pathogenic organisms infecting the body, the body's innate immune response is first activated. The activation of innate immune response mainly depends on the recognition of pathogen-specific components by pattern recognition receptors (PRRs) on the surface of immune cells. The PRRs discovered so far are mainly divided into three categories: 1. TLRs (Toll-like receptors); 2. RLHs (RIG-I-like helicases); 3. NLRs (Nucleotide-oligomerization domain-like receptors). In the process of antiviral innate immune response, viral components such as DNA or RNA can be recognized by TLRs and RLHs of immune cells, thereby activating downstream signaling pathways, prompting immune cells to activate and express...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/118C12Q2600/158
Inventor 周烨曹雪涛侯晋
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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