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Colon cancer diagnostic kit and preparation method thereof

A diagnostic kit and gene technology, applied in the field of biomedicine, can solve problems such as inconsistent analysis results

Inactive Publication Date: 2014-11-12
FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, differences in different experimental platforms and samples lead to many inconsistencies in the analysis results

Method used

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  • Colon cancer diagnostic kit and preparation method thereof
  • Colon cancer diagnostic kit and preparation method thereof
  • Colon cancer diagnostic kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Screening for genes associated with colon cancer

[0043] 1.1 NCBI GEO (Gene Expression Omnibus) data retrieval

[0044] The GEO (Gene Expression Omnibus) database is developed and maintained by NCBI (National Center for Biotechnology Information). The GEO database is the largest database of gene expression data. SAGE (Serial Analysis of Gene Expression) data, SARST (Serial Analysis of Ribosomal Sequence Tags) data, MS (Mass Spectrometry) data, proteome data and next-generation high-throughput sequencing data (MPSS, massively parallel sequencing technology), etc.

[0045] a. Search keywords: ("colorectal cancer"[MeSH Terms]OR"colorectal cancer"

[0046] [All Fields]) AND "Homo sapiens"[porgn]

[0047] b. Sample selection strategy in the study:

[0048] Limit the research type to "expression profiling by array" Data sets that meet the following criteria will be included in our research: ①The selected data sets must be genome-wide expression mRNA transcriptom...

Embodiment 2

[0053] Embodiment 2QPCR verifies the relationship between candidate genes and colon cancer

[0054] Based on the results of meta-analysis of colon cancer gene chip data, according to the size of the P value, we selected the FOXD4L4 gene (Meta analysis showed that the expression of FOXD4L4 gene was down-regulated in colon cancer tissues compared with normal tissues) for verification. Collect 50 colon cancer tissue samples and 50 normal tissue samples at the same time, and use fluorescent real-time quantitative PCR to carry out classical molecular biology experiment verification (qRT-PCR). The specific operation steps are as follows:

[0055] (1) RNA extraction

[0056] After collecting the samples, freeze them in liquid nitrogen. After taking them out, put the tissue into a pre-cooled mortar for grinding. After the tissue sample is powdered:

[0057] ① Add Trizol and store at room temperature for 5 minutes;

[0058] ② Add 0.2ml of chloroform, vibrate the centrifuge tube vigor...

Embodiment 3

[0067] The preparation of embodiment 3 colon cancer diagnosis kits

[0068] According to the correlation between the FOXD4L4 gene and colon cancer, whether colon cancer occurs can be diagnosed by detecting the expression of the FOXD4L4 gene. Accordingly, the present invention provides a kit for diagnosing colon cancer based on detecting the expression of the FOXD4L4 gene. The diagnostic kit The components are as follows: SYBR Green polymerase chain reaction system; primer pair for amplifying FOXD4L4 gene and GAPDH gene. The forward primer sequence for amplifying the FOXD4L4 gene is 5'-GAGGCGAGACAGCAGTTCCTAGAG-3', the reverse primer sequence is 5'-GATGTACGAGTAGGGGGGCTTTG-3'; the forward primer sequence for amplifying GAPDH is 5'-TTTAACTCTGGTAAAGTGGATAT-3', the reverse The primer sequence was 5'-GGTGGAATCATATTGGAACA-3'. The SYBR Green polymerase chain reaction system includes PCR buffer, dNTPs, and SYBR Green fluorescent dye. PCR buffer composition: 25mM KCL, 2.5mM MgCL 2 , 2...

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Abstract

The invention discloses an application of an FOXD4L4 gene in preparing a kit for diagnosing colon cancer. By adopting NCBI GEO data retrieval, genome mRNA expression chip data of five colon cancer cases which meet the requirements and normal people are brought into the study range, Meta analysis on the chip data shows that the FOXD4L4 gene is a difference remarkable gene, compared with normal tissue, the expression of the FOXD4L4 gene in colon cancer tissue is down-regulated, and the conclusion is verified by using a fluorescence real-time quantitative PCR method. On such a basis, the invention further discloses a kit for diagnosing colon cancer and a method for diagnosing colon cancer by using the kit. On one hand, a novel idea is provided to study the molecular mechanism of colon cancer, and on the other hand, a relatively sensitive diagnosis kit is provided for primary colon cancer diagnosis.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a colon cancer diagnostic kit and the application of FOXD4L4 gene in preparing the colon cancer diagnostic kit. Background technique [0002] Colon cancer is the third most common malignant tumor in both men and women. According to the 2013 American Cancer Statistics Report, there were 103,170 new cases of colon cancer in the United States in 2012. In my country, with the changes in lifestyle and eating habits in recent years, the incidence and death of colon cancer have risen rapidly. According to statistics in 2011, colon cancer ranks third in the incidence of malignant tumors in my country and is the fourth leading cause of cancer. The morbidity and mortality of malignant tumors are close to the world level at the same time, becoming a public health problem that seriously threatens the life and health of the population. The pathogenesis of colon cancer has not yet been fully elucidated...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/158
Inventor 肖文华屈雪玲王硕李小梅董伟伟赵慧霞
Owner FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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