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Application of LINC00266-1 polypeptide as solid tumor marker

A solid tumor and polypeptide technology, applied in the application field of solid tumor markers, can solve the problems of lack of tumor monitoring indicators, and achieve high mortality, short survival period, and the effect of judging the degree of malignancy and metastasis tendency

Active Publication Date: 2019-03-26
THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The lack of effective tumor monitoring indicators is an important reason

Method used

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  • Application of LINC00266-1 polypeptide as solid tumor marker
  • Application of LINC00266-1 polypeptide as solid tumor marker
  • Application of LINC00266-1 polypeptide as solid tumor marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Prepare the anti-LINC00266-1 polypeptide antibody, and detect the titer of the prepared anti-LINC00266-1 polypeptide antibody, the specific operation process is as follows:

[0053] (1) Synthesize the designed epitope peptide (sequence shown in SEQ ID NO: 2) against the LINC00266-1 polypeptide by chemical synthesis (Gill Biochemical (Shanghai) Co., Ltd.), and then synthesize the epitope peptide The immune antigen is coupled with keyhole limpet hemocyanin, and the immune antigen is separated and purified by high pressure liquid chromatography (HPLC) to make the purity >85%.

[0054] Take 2 healthy New Zealand rabbits weighing no less than 2kg and name them Rabbit No. 1 and Rabbit No. 2 respectively. 2 to 3 ml of blood from the above-mentioned rabbit ear arteries were collected, and serum was obtained by separation, which was used as negative control serum.

[0055] (2) New Zealand rabbits No. 1 and No. 2 were injected with the immune antigens separated and purified by H...

Embodiment 2

[0065] The specific steps of the specific detection experiment of anti-LINC00266-1 antibody to LINC00266-1 polypeptide are as follows:

[0066] The first step is to construct the LINC00266-1-GFP fusion gene expression vector. First, site-directed mutation of the wild-type GFP (GFPwt) gene translation promoter ATGGTG sequence to ATTGTT (GFPmut). Therefore, the translation of the GFP gene is destroyed, and the translation promoter ATG on the LINC00266-1 gene is used for translation, thereby expressing and producing the LINC00266-1-GFP fusion protein. After transfecting HeLa cells (ATCC, Number: CCL-2) with liposome Lipo2000 (Life technology) for 24 hours, the cells were collected and lysed to prepare protein samples. Using the anti-GFP antibody and the anti-LINC00266-1 polypeptide antibody of the present invention respectively, the expression of LINC00266-1-GFP fusion protein was detected by Western blotting method. The result is as figure 1 As shown in a, the molecular weigh...

Embodiment 3

[0069] Using the Western blotting method, the anti-LINC00266-1 antibody prepared by the present invention was used to detect the expression level of the LINC00266-1 polypeptide in colon cancer cells with different invasion and metastasis potentials from the same source. The specific process is as follows:

[0070] Two pairs of colon cancer cells SW480 and SW620 (ATCC, Number: CCL-228 and CCL-227, respectively), HTC-116 (ATCC, Number: CCL-247) and HTC-116high (this cell Established by the previous screening in our laboratory), the cells were lysed to prepare protein samples. Using the Western blotting method, the anti-LINC00266-1 polypeptide antibody prepared by the present invention was used to detect the difference in the level of LINC00266-1 polypeptide between the paired cells. The result is as figure 2 As shown, the level of LINC00266-1 polypeptide was significantly up-regulated in colon cancer cells SW620 and HTC-116high with high invasion and metastasis potential. In ...

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Abstract

The invention relates to the field of diagnosis and treatment of cancers and discloses an application of LINC00266-1 polypeptide as a solid tumor marker. Measurement of the LINC00266-1 polypeptide canbe used for detecting or diagnosing tumors or evaluating and monitoring prognosis of a patient.

Description

technical field [0001] The present invention relates to the field of diagnosis and treatment of cancer, in particular, relates to the application of LINC00266-1 polypeptide as a solid tumor marker. Background technique [0002] Cancer is the second leading cause of death in the world, killing approximately 8.8 million people each year. Globally, nearly one in six deaths is due to cancer. In recent years, with the improvement of people's material living standards, accelerated pace of life, changes in dietary structure, aging population, environmental pollution and other reasons, the incidence of solid malignant tumors has been increasing year by year, and it is showing a younger trend. [0003] Although the 5-year survival rate of patients with early solid malignant tumors can reach more than 80%, the rate of patients seeing a doctor is low because early solid malignant tumors have no specific symptoms and signs. Coupled with the lack of convenient and effective screening m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47G01N33/574
CPCC07K14/47G01N33/57484G01N2333/47
Inventor 晏光荣朱嵩王继重
Owner THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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