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Molecular markers for detecting lung metastasis of minimal osteosarcoma and application thereof

An osteosarcoma and diagnostic marker technology, applied in the field of clinical medicine, can solve the problems of difficult early diagnosis of osteosarcoma lung metastasis, less than 20% survival rate, poor prognosis, etc.

Pending Publication Date: 2019-12-03
THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Abstract
  • Description
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Problems solved by technology

[0003] Osteosarcoma with pulmonary metastases is challenging to manage and often has a poor prognosis, even with a multidisciplinary approach
The current treatment mainly adopts surgical resection of metastases and increasing the dose of chemotherapy to control metastases. However, its 5-year survival rate is less than 20%.
Moreover, there are no molecular markers that clearly predict osteosarcoma metastasis, these factors make the early diagnosis of osteosarcoma lung metastasis particularly difficult

Method used

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  • Molecular markers for detecting lung metastasis of minimal osteosarcoma and application thereof
  • Molecular markers for detecting lung metastasis of minimal osteosarcoma and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 ctDNA Mutation Gene Detection and Prognosis Judgment of Osteosarcoma Patient 1

[0022] 1. Extraction of blood DNA: (1) Take 10ml of peripheral blood from the patient. Add cell washing solution to the whole blood sample, wherein the volume ratio of whole blood sample to cell washing solution is 5:14, mix the sample upside down, incubate at room temperature for 5-10 minutes, centrifuge at 13000-16000g for 1 minute at room temperature, discard the supernatant , to obtain residual cell clusters; (2) digestion and precipitation: get 20~30ul of the residual cell clusters obtained in the above-mentioned steps (1), and then add 15ul of sodium dodecyl sulfate SDS with a volume fraction of 10%, 15ul of 8M SDS with a concentration of 8M Ammonium acetate, 600ul of guanidine hydrochloride with a concentration of 5M, 3ul of proteinase K with a concentration of 20mg / ml, after 15 to 30 minutes in a water bath at 60°C, cool to room temperature in an ice bath, and then centrif...

Embodiment 2

[0030] Example 2, ctDNA mutation gene detection and prognosis judgment of patient 2

[0031] 1. Extraction of blood DNA: (1) Take 10ml of peripheral blood from the patient. Add cell washing solution to the whole blood sample, wherein the volume ratio of whole blood sample to cell washing solution is 5:14, mix the sample upside down, incubate at room temperature for 5-10 minutes, centrifuge at 13000-16000g for 1 minute at room temperature, discard the supernatant , to obtain residual cell clusters; (2) digestion and precipitation: take 20~30ul of the residual cell clusters obtained in the above-mentioned steps (1), and then add 15ul of sodium dodecyl sulfate SDS with a volume fraction of 10%, 15ul of 8M SDS with a concentration of 8M Ammonium acetate, 600ul of 5M guanidine hydrochloride, 3ul of 20mg / ml proteinase K, after 15-30 minutes in a water bath at 60°C, cool to room temperature in an ice bath, and then centrifuge at 13000-16000rpm / min at 4-8°C 2~3min, obtain the centrifug...

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Abstract

The invention provides an application of blood circulation ctDNA mutation as a diagnostic marker in preparation of a diagnostic kit for detecting osteosarcoma metastasis. The blood circulation ctDNA can provide comprehensive description of tumor genomes, because ctDNA is released from multiple different tumor regions or multiple lesions, and ctDNA analysis can discover somatic mutations that cannot be found in tissue biopsy. The occurrence of the ctDNA is a powerful marker for cancer recurrence and poor prognosis, and the ctDNA has a prognosis prediction value.

Description

technical field [0001] The invention relates to the technical field of clinical medicine, in particular to a group of molecular markers for detecting micro-osteosarcoma lung metastasis and their application. Background technique [0002] Osteosarcoma is the third most malignant bone tumor in children and adolescents, and it is very prone to lung metastasis. After standardized neoadjuvant chemotherapy and extensive resection, the five-year survival rate is close to 60%. Once lung metastasis occurs, the 5-year survival rate is often lower than 20%. Therefore, lung metastasis is the main factor hindering the long-term survival of osteosarcoma patients. The one-child family has become the mainstream family system in China. Osteosarcoma is prone to occur in adolescents and the mortality rate is high, which will pose a huge threat to the above-mentioned families. If an early warning system for lung metastasis of osteosarcoma can be established, the progression of the disease ca...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/156C12Q2600/118C12Q2600/106
Inventor 王晋徐怀远陈宏敏邓创忠许衍扬宋怿江唐清连卢金昌朱小军宋国徽
Owner THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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