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miRNAs as targets for diagnosis and treatment of osteosarcoma

A technology for osteosarcoma and a diagnostic tool, which is applied in the field of diagnosis and treatment of osteosarcoma by miR-4714-3p, can solve problems such as abnormal gene expression and induced diseases, and achieve the effect of small diagnosis in time, reduce mortality, and realize early diagnosis

Active Publication Date: 2018-09-18
杨祚璋
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The abnormal expression of miRNA directly leads to the abnormal expression of some genes related to the occurrence of diseases, which induces the occurrence of diseases.

Method used

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  • miRNAs as targets for diagnosis and treatment of osteosarcoma
  • miRNAs as targets for diagnosis and treatment of osteosarcoma
  • miRNAs as targets for diagnosis and treatment of osteosarcoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 High-throughput sequencing screening of miRNAs associated with osteosarcoma

[0038] 1. Sample collection

[0039] 5 patients with primary osteosarcoma and 5 normal people. Subjects were required to fast for at least 12 hours, and at room temperature from 7:00 to 8:00 in the morning of the next day, 10ml of venous blood was drawn into EDTA anticoagulant tubes to extract PBMCs from peripheral blood mononuclear cells, and 1ml of Trizol was added Reagents (Invitrogen Company), mixed thoroughly, and samples were stored at -80°C for RNA extraction. All blood samples and pathological results should be authentic and reliable, the study was approved by the ethics committee, and the patients gave informed consent.

[0040] 2. RNA extraction from blood monocytes

[0041] Thaw the cells in step (1), add about 1 / 5 of the volume of chloroform, mix well by inverting up and down for about 1 minute, and let stand at room temperature for 5 minutes. After centrifugation at ...

Embodiment 2

[0052] Example 2 QPCR verification of differentially expressed miR-4714-3p

[0053] 1. According to the high-throughput sequencing results in Example 1, miR-4714-3p was selected for large-sample QPCR verification. According to the sample collection method in Example 1, 50 cases of normal people and 50 cases of osteosarcoma patients were selected to separate and store blood mononuclear cells.

[0054] 2. The RNA extraction process is the same as in Example 1.

[0055] 3. Reverse transcription: mix 10pg-1μg total RNA template with 2μl 10* buffer, 2μl dATP (10mM), 0.5μl polyA polymerase, 0.5μl ribonuclease (RNase) inhibitor and ribonuclease free water (RNase free water ) were mixed, the final volume was 20 μl, and incubated at 37° C. for 1 h. Then add 1 μl 0.5 μg / μl Oligo(dT) specific RT primer to the reaction tube, incubate at 70°C for 5 minutes and immediately incubate on ice for at least 2 minutes to break the secondary structure of RNA and primers. Finally, mix the above 2...

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Abstract

The invention discloses application of miR-4714-3p regarding as an osteosarcoma diagnostic marker. The high-throughput sequencing and QPCR (Quantitative Polymerase Chain Reaction) experiment show the content of the miR-4714-3p is significantly different in bloods of average persons and osteosarcoma patients, so that the miR-4714-3p can be regarded as a molecular marker for diagnosing osteosarcoma so as to be used for developing a product for diagnosing osteosarcoma. The diagnostic product has the following advantages of noninvasiveness, quickness, sensitiveness, and clinically wide application prospect.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of miR-4714-3p in the diagnosis and treatment of 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 low cured survival rate. X-ray, tomography, nuclear magnetic resonance, angiography and dynamic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 杨祚璋张瞾昕夏俊峰李惠玲张娅李素
Owner 杨祚璋