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Application of COCH gene and its expression product

A gene and gene chip technology, applied in the field of application of COCH gene and its expression products in the diagnosis of liver cancer, can solve the problems of unclear relationship and achieve accurate diagnosis

Inactive Publication Date: 2012-09-26
CHINESE NAT HUMAN GENOME CENT AT SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the expression profile of human COCH gene in tumors and its relationship with tumorigenesis are still unclear, and further research is needed

Method used

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  • Application of COCH gene and its expression product
  • Application of COCH gene and its expression product
  • Application of COCH gene and its expression product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 RT-PCR assay to detect the expression of COCH gene in liver cancer tissue

[0025] Reverse transcription-polymerase chain reaction (RT-PCR) was used to examine the expression of COCH gene in liver cancer tissues. RT-PCR refers to a method that combines a reverse transcription (Reverse Transcription; RT) reaction and a PCR (Polymerase Chain Reaction) reaction. RT-PCR combines RNA-templated cDNA synthesis with PCR to provide a fast and sensitive method for analyzing gene expression. RT-PCR is used to detect or quantify expression information. RT-PCR is more sensitive and easier to operate than other RNA analysis techniques including Northern blotting, RNase protection analysis, in situ hybridization and S1 nuclease analysis. The template for RT-PCR can be total RNA or poly(A) selective RNA. The reverse transcription reaction can be initiated with a reverse transcriptase, random primers, oligo(dT), or gene-specific primers. RT-PCR can be performed in a one-st...

Embodiment 2

[0049] Example 2 Realtime-PCR assay to detect the expression of COCH gene in liver cancer tissue

[0050] Relative quantitative method was used to detect the expression difference of STC2 gene in liver cancer samples (Thermal Cycler Dice TM RealTime System TP800, Takara; Premix Ex Taq TM , Takara).

[0051] The fluorescent quantitative PGR (real-time PGR) reaction system is as follows: total volume 20 μL, SYBR Premix Ex Taq 10 μL, upstream and downstream primers (10 μmol / L) 0.4 μL each, cDNA 1 μL, ddH2O 8.2 μL, mix the reagents, centrifuge and put The amplification reaction was carried out in a PGR instrument. The reaction conditions were denaturation at 95°C for 5 seconds, annealing and extension at 68°C for 30 seconds, 40 cycles. The instrument was operated according to the manufacturer's instructions. The housekeeping gene β-actin was used as an internal reference.

[0052] The primers designed in this example are as follows:

[0053] COCH-F: 5'-ACAGGAAAAGCCTTGAAGCA...

Embodiment 3

[0058] Example 3 PGR experiment detects the expression of COCH gene in normal human tissue

[0059] The expression of COCH gene in normal adult tissues was detected by conventional PGR method. Experimental results show that COCH is expressed in embryonic liver but not in mature liver (see image 3 ), further indicating that the expression of COCH gene can be used for accurate diagnosis of liver cancer.

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PUM

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Abstract

The invention discloses application of a COCH gene and its expression product in preparing liver cancer diagnosis products. The COCH gene and its expression product can be taken as a specific marker gene of liver cancer diagnosis, thus making liver cancer diagnosis more accurate and rapid.

Description

technical field [0001] The invention relates to the application of a gene, in particular to the application of a COCH gene and its expression product in the diagnosis of liver cancer. Background technique [0002] Liver cancer is a serious threat to the life and health of our people. The diagnosis of liver cancer, especially early diagnosis is the key to clinical diagnosis, treatment and prognosis. At present, the qualitative diagnosis of liver cancer in my country is still based on the detection of serum AFP (alpha-fetoprotein), which presents the following characteristics: more than 60% of liver cancer cases have serum AFP > 400 μg / L; currently there is no other tumor marker specificity comparable to Comparable to AFP; AFP detection is less dependent on imaging equipment and new technologies. AFP is currently the most widely used liver cancer tumor marker in the world. It has been used for decades. Its sensitivity is 40% to 65%, and its specificity is 76% to 96%. Such ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/574C12N15/11
Inventor 韩泽广阎冰邓庆
Owner CHINESE NAT HUMAN GENOME CENT AT SHANGHAI