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New drug target for 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: 2015-05-27
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, using the above clinical methods to diagnose osteosarcoma often leads to delays in the patient's condition and misses the best treatment period.

Method used

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  • New drug target for osteosarcoma
  • New drug target for osteosarcoma
  • New drug target for 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 the previous high-throughput transcriptome deep sequencing, the C8orf59 gene (which is expressed in bone and flesh 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 min...

Embodiment 3

[0094] Example 3 Expression changes of C8orf59 protein in immunoassay samples

[0095] 1. Acquisition of antigenic protein

[0096] Expression by genetic engineering: the cDNA sequence of the human C8orf59 gene can be obtained from the Genbank database, the coding frame can be obtained by PCR amplification, inserted into the prokaryotic or eukaryotic expression vector, the C8orf59 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 C8orf59 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...

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Abstract

The invention discloses application of C8orf59 gene in preparing drugs for diagnosing and treating osteosarcoma. The experiment proves that the expression level of the C8orf59 gene in osteosarcoma tissues is obviously higher than that in normal tissues, and therefore, the C8orf59 gene can be used as a specific marker for diagnosing osteosarcoma, so that the osteosarcoma diagnosis is more accurate and faster. The expression of the C8orf59 gene in the osteosarcoma cell line can be disturbed to obviously inhibit the cell proliferation, migration, invasion and tumor formation capacity and accelerate apoptosis. Therefore, the C8orf59 gene has potential application value for gene therapy of osteosarcoma, and is hopeful to be used for treating osteosarcoma and developing new anti-osteosarcoma drugs on such basis, thereby making contribution for preventing and treating osteosarcoma in clinic.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a new drug target for osteosarcoma, in particular to the application of C8orf59 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 rat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68C12N15/11A61K48/00A61K31/7088A61P35/00
Inventor 杨承刚孙耀兰边洋
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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