Application of WWP1 gene in preparing products for diagnosing osteosarcoma and drugs for treating osteosarcoma

A technology of osteosarcoma and osteosarcoma cells, applied in the field of biomedicine, can solve the problems of patients' condition delay and missing the best treatment period

Active Publication Date: 2015-05-13
GUAN BOJIAN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, using the above clinical methods to diagnose osteosarcoma often ...

Method used

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  • Application of WWP1 gene in preparing products for diagnosing osteosarcoma and drugs for treating osteosarcoma
  • Application of WWP1 gene in preparing products for diagnosing osteosarcoma and drugs for treating osteosarcoma
  • Application of WWP1 gene in preparing products for diagnosing osteosarcoma and drugs for treating osteosarcoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Screening for genes related to osteosarcoma

[0056] 1.1 Sample collection

[0057] 8 samples of normal bone tissue and osteosarcoma tissue were collected respectively.

[0058] 1.2 RNA sample preparation and quality analysis

[0059] 1.2.1 RNA sample preparation

[0060] After RNA extraction from normal bone tissue and osteosarcoma tissue, the quality of the RNA sample was judged by agarose gel electrophoresis, and the qualified ones could be used for further transcriptome analysis.

[0061] 1.2.2 Quality analysis of RNA samples (NanoDrop1000 spectrophotometer)

[0062] NanoDrop1000 spectrophotometer detects RNA samples, and the sample requirements for RNA-seq sequencing: OD260 / OD280 is 1.8-2.2.

[0063] 1.2.3 Quality analysis of RNA samples (Agilent Technologies 2100 Bioanalyzer)

[0064] Agilent Technologies 2100 Bioanalyzer detects the quality of RNA samples, observes that the main bands of 28S rRNA and 18S rRNA are obvious, there is no degradation, t...

Embodiment 2

[0074] Example 2 Large sample PCR verification of the relationship between candidate genes and osteosarcoma

[0075] Considering the gene that has not been studied on the relationship between this gene and osteosarcoma in the prior art as a candidate gene, and considering the results of high-throughput transcriptome deep sequencing in the previous stage, the WWP1 gene (expressed in bone and muscle cells) was selected according to the size of the P value. up-regulated in tumor tissue) for verification. Collect 30 osteosarcoma tissues and 20 normal bone tissue samples at the same time, and use PCR to carry out classical molecular biology experiment verification. The specific operation steps are as follows:

[0076] 1. RNA extraction

[0077] 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:

[0078] ① Add Trizol and store at room temperature for 5 minut...

Embodiment 3

[0094] Example 3 Expression changes of WWP1 protein in immunoassay samples

[0095] 1. Acquisition of antigenic protein

[0096]Expression by genetic engineering: the cDNA sequence of the human WWP1 gene can be obtained from the Genbank database, the coding frame can be obtained by PCR amplification, inserted into the expression vector of prokaryotes or eukaryotes, WWP1 protein can be expressed, and the purification system of the expression product can be followed by genetic engineering Purify protein.

[0097] 2. Antibody Preparation

[0098] Antibodies can be prepared in several ways:

[0099] (1) Cell fusion method: immunize animals (including rabbits, goats, etc.) with the WWP1 protein prepared above, obtain spleen cells, fuse with myeloma cells, and prepare monoclonal antibodies according to conventional monoclonal antibody preparation techniques.

[0100] (2) Using the phage surface display library, cloning the spleen IgG variable region of the immunized animal and ex...

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Abstract

The invention discloses an application of a WWP1 gene. The WWP1 gene can be used for preparing products for diagnosing osteosarcoma and is used as a specific marker gene for diagnosing osteosarcoma so that the osteosarcoma is more accurately and quickly diagnosed. The products for diagnosing osteosarcoma comprise products for diagnosing osteosarcoma by virtue of RT-PCR, real-time quantitative PCR, immunodetection, in-situ hybridization or gene chips. The WWP1 gene disclosed by the invention can also be used for preparing drugs for treating osteosarcoma which provides novel therapeutic targets and effective novel drugs for prevention and treatment of osteosarcoma.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a new application of WWP1 gene, in particular to the application of WWP1 gene in preparing products for diagnosing osteosarcoma and preparing drugs for treating osteosarcoma. Background technique [0002] Osteosarcoma is a malignant tumor derived from mesenchymal tissue, and its main pathogenic feature is that the proliferating tumor cells in the body directly form immature bone or osteoid tissue. It is the most common primary malignant tumor of the human skeletal system. Typical osteosarcoma is a rare (accounting for 0.2% of all malignant tumors) highly carcinogenic malignant tumors, and its incidence rate is about three cases per million people per year. Osteosarcoma mainly occurs in longer bones and to a lesser extent in soft tissues. Its main onset population is concentrated in adolescents aged 10 to 20, and has the characteristics of high incidence, high early metastasis rate, and ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574C12N15/113A61K48/00A61K31/713A61K39/00A61P35/00
Inventor 杨承刚边洋孙耀兰
Owner GUAN BOJIAN BIOTECH CO LTD
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