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Application of miRNA as biomarker in preparation of osteosarcoma detection preparation

A technology for biomarkers and osteosarcoma, applied in the field of biomedicine, can solve the problems of mottled information, interference, and diverse sources, and achieve the effect of not easy to coagulate, eliminate influence, and pure ingredients

Inactive Publication Date: 2021-12-03
SHANGHAI KINGMED DIAGNOSTICS INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, although blood samples contain a lot of information about general conditions, liquid biopsy by extracting miRNA in blood still has some inherent defects: first, blood collection is invasive and causes discomfort to patients; second, this Direct detection of naked DNA or miRNA in blood will also cause different degrees of loss and degradation; third, blood components are complex, sources are diverse, and information is mottled, which may cause non-specificity and hydrodynamic effects during the detection and analysis process. interference; fourth, blood is prone to coagulation, and anticoagulant treatment is required to carry out the next test, and anticoagulant treatment itself may have a certain impact on the test results; fifth, a large number of cells in the blood have a few seconds A half-life of several weeks or more than a month may also affect the results of the analysis

Method used

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  • Application of miRNA as biomarker in preparation of osteosarcoma detection preparation
  • Application of miRNA as biomarker in preparation of osteosarcoma detection preparation
  • Application of miRNA as biomarker in preparation of osteosarcoma detection preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Gene chip high-throughput screening of miRNA expression differences in salivary exosomes of osteosarcoma patients and normal people

[0048] In this example, the miRNAs in the salivary exosomes of 20 patients with osteosarcoma and the miRNAs in the salivary exosomes of 20 normal volunteers were first extracted, and then the osteosarcoma patients and normal volunteers were screened by high-throughput gene chips Differences in miRNA expression. Specifically include the following steps:

[0049] 1. Collection, preservation and transportation of saliva

[0050] This example involves the saliva samples of 20 patients with osteosarcoma in Changzheng Hospital Affiliated to Second Military Medical University and 20 normal volunteers in the hospital, and other factors such as age, difference between men and women, and smoking or not between the two groups are strictly controlled. Collect, store and transport according to the following standards and procedures:

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Embodiment 2

[0069] Example 2 Q-PCR verification hsa-miR-34a, hsa-miR-1270, hsa-miR-154, hsa-miR-329 detection of osteosarcoma

[0070] Extract the miRNAs in the salivary exosomes of 100 patients with osteosarcoma in Changzheng Hospital and the miRNAs in the salivary exosomes of 40 normal people, using the hsa-miR-34a, hsa-miR-1270 screened in Example 1, hsa-miR-154 and hsa-miR-329 were verified by Q-PCR and relative quantification by △△CT method. Specifically include the following steps:

[0071] 1. Reverse transcription of miRNA in salivary exosomes into cDNA

[0072]miRNAs in salivary exosomes of 100 patients with osteosarcoma were reverse-transcribed into cDNA by tailing method and stem-loop method, and miRNAs in salivary exosomes of 40 normal people were reverse-transcribed into cDNA, Use mature kits on the market such as All-in-OneTMmiRNA qRT-PCRDetection (QP015, QP016) from Funeng Company, operate according to the instructions, and prepare a 25uL reverse transcription reaction s...

Embodiment 3

[0084] Example 3 Correlation analysis between miRNA in salivary exosomes and miRNA in osteosarcoma cancer tissue

[0085] hsa-miR-34a, hsa-miR-1270, hsa-miR-154, hsa-miR-329 in cancer tissues of patients with osteosarcoma and hsa-miR-34a, hsa-miR in salivary exosomes of patients with osteosarcoma -1270, hsa-miR-154, hsa-miR-329, for correlation analysis, the results are as follows Figure 2-5 shown.

[0086] Figure 2-5 The results showed that the correlations were p=0.0201, p=0.0130, p=0.0322, p=0.0039, indicating that hsa-miR-34a, hsa-miR-1270, hsa-miR-154 in salivary exosomes of patients with osteosarcoma , hsa-miR-329 have good consistency with hsa-miR-34a, hsa-miR-1270, hsa-miR-154, hsa-miR-329 in osteosarcoma cancer tissue, miRNA in salivary exosomes It can reflect the expression level of miRNA in cancer tissue.

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Abstract

The invention discloses application of miRNA in preparation of an osteosarcoma detection preparation. The miRNA is one or more of hsa-miR-34a, hsa-miR-1270, hsa-miR-154 and hsa-miR-329. The invention discovers that the hsa-miR-34a, the hsa-miR-1270, the hsa-miR-154 and the hsa-miR-329 are significantly up-regulated in the saliva exosome of an osteosarcoma patient and have high consistency with osteosarcoma tissues, so that one or more of the four miRNAs can be adopted as an osteosarcoma marker and can be used in the preparation of the osteosarcoma detection preparation.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and more specifically, the invention relates to the application of multiple miRNAs as biomarkers in the preparation of osteosarcoma detection preparations for non-disease diagnosis methods. Background technique [0002] Osteosarcoma is the most common malignant bone tumor in children and adults. Among primary bone tumors, the incidence rate of osteosarcoma is second only to plasma cell myeloma, and it is more common in young people aged 10-25, male and female. The patient ratio is approximately 1.5:1. Osteosarcoma has a high degree of malignancy, very rapid growth, early metastasis, and strong invasiveness. About 80% of patients have metastases to the lung or other parts when they are diagnosed clinically. In the past, the prognosis of patients with osteosarcoma was extremely poor, and the 5-year survival rate was only about 20%. Although great progress has been made in surgery and chemother...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 陈天睿王阳王迪周旺曹昊李博谢江涛王祥敏黄健刘贞陈平张辛宁严佳杰王波
Owner SHANGHAI KINGMED DIAGNOSTICS INST