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Method and kit for prognosis judgment of colon cancer

A prognostic judgment, colon cancer technology, applied in the direction of disease diagnosis, biological testing, material testing products, etc., can solve the problems of recurrence, tumor metastasis, etc., achieve high mortality, short survival period, improve the effect of positive coincidence rate

Active Publication Date: 2016-10-26
THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, even though the TNM stage of many colon cancer patients belongs to the early stage, the tumor still metastasizes and recurs, which shows that some colon cancers that have not metastasized through the current clinical diagnosis and treatment technology still have the potential of metastasis and recurrence

Method used

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  • Method and kit for prognosis judgment of colon cancer
  • Method and kit for prognosis judgment of colon cancer
  • Method and kit for prognosis judgment of colon cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] In order to illustrate the preparation of anti-HOXB-AS3 antibody and the titer of the prepared anti-HOXB-AS3 antibody, the specific operation process is as follows:

[0048] (1) Chemically synthesize the designed antigenic epitope peptide against HOXB-AS3 polypeptide (Gill Biochemical (Shanghai) Co., Ltd.), and then the synthesized antigenic epitope peptide is coupled with keyhole limpet hemocyanin as the immune antigen. Antigens were purified by high pressure liquid chromatography (HPLC).

[0049] Take 2 New Zealand rabbits weighing not less than 2kg and name them Rabbit 1 and Rabbit 2 respectively. 2-3 ml of blood from the above-mentioned rabbit ear arteries was collected, and the serum was separated to obtain serum, which was the negative control serum.

[0050] (2) The above-mentioned purified immunizing antigens are injected at multiple points of immunizing antigens. The specific process is as follows:

[0051] First, dissolve the epitope peptide coupled to keyh...

Embodiment 2

[0060] The specific steps for the specific detection of the HOXB-AS3 polypeptide by the anti-HOXB-AS3 antibody are as follows:

[0061] Construct HOXB-AS3-GFP fusion gene expression vector. Among them, the wild-type GFP (GFPwt) gene translation promoter ATGGTG was mutated into ATTGTT (GFPmut), thereby destroying the independent translation of the GFP gene, and using the translation promoter ATG on the HOXB-AS3 gene for translation to produce HOXB-AS3 AS3-GFP fusion protein. After transfecting HeLa cells (ATCC, Number: CCL-2) with liposome Lipo2000 (Lifetechnology) for 24 hours, use the anti-GFP antibody and the anti-HOXB-AS3 polypeptide antibody prepared above to detect HOXB by immunoblotting - Expression of AS3-GFP protein. The result is as figure 1 As shown in a, both the anti-GFP antibody and the anti-HOXB-AS3 antibody can specifically detect the expression of the HOXB-AS3-GFP fusion protein. However, the protein band cannot be detected by corresponding Western blot det...

Embodiment 3

[0064] Using the method of immunoblotting, using the anti-HOXB-AS3 antibody prepared above to detect the expression level of the HOXB-AS3 polypeptide in colon cancer cells with different invasion and metastasis potentials from the same source, the specific steps are as follows:

[0065] Using this anti-HOXB-AS3 antibody, two pairs of colon cancer cells SW480 and SW620 (ATCC, Number: CCL-228 and CCL-228 respectively) and HTC-116 (ATCC, Number: CCL-247) and HTC-116 high (This cell was established by the previous screening of our laboratory) The difference in the level of HOXB-AS3 polypeptide, found that the level of HOXB-AS3 polypeptide in high invasion and metastasis potential colon cancer cells SW620 and HTC-116 high Medium and low, see the results figure 2 , to detect β-actin as an internal reference.

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Abstract

The invention provides a method for prognosis judgment of colon cancer. The method comprises the step of detecting the amino acid sequence of a polypeptide, wherein the amino acid sequence is the sequence shown in SEQ NO.1 or a sequence with 99% of display content identical with the display content of the sequence shown in SEQ NO.1. A novel marker, namely HOXB-AS3 polypeptide, for prognosis evaluation and judgment of colon cancer is also provided. Clinical study proves that low expression or expression failure of the HOXB-AS3 polypeptide in colon cancer tissue is positively related to poor prognosis of colon cancer patients, and patients with low HOXB-AS3 polypeptide level in colon cancer tissue have higher death rate and shorter lifetime. Therefore, by using the HOXB-AS3 polypeptide as the biomarker, the positive coincidence rate is effectively increased when the kit is clinically applied to prognosis judgment of colon cancer. Furthermore, by detecting the level of the HOXB-AS3 polypeptide in a biological sample tissue slice, generation, malignancy grade and metastatic potential of colon cancer can be judged easily.

Description

technical field [0001] The invention relates to the field of diagnosis and treatment of tumors, in particular to a method and a kit for judging the prognosis of colon cancer. Background technique [0002] Colon cancer is a common malignant tumor of the digestive system in the world, and its incidence rate ranks third among all kinds of tumors. There are about 1.2 million new cases worldwide every year, and the annual death toll is as high as 600,000. In recent years, with the improvement of people's living standards, changes in lifestyle and diet structure, and the aggravation of population aging, the incidence of colon cancer has increased year by year in my country, and it is showing a younger trend. It ranks 3rd and 5th in the incidence and mortality of tumors in my country. [0003] The 5-year survival rate of patients with early localized colon cancer can reach 90%, but less than 40% of patients with colon cancer are confirmed in the early stage. When most colon cance...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/574
CPCG01N33/57419G01N33/57484G01N33/68G01N2333/47G01N2800/52
Inventor 晏光荣黄金舟陈敏
Owner THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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