Application of hsa_circ_0000284 in preparation of neuroblastoma prognostic preparation

A neuroblastoma, prognosis technology, applied in the field of molecular biology, can solve the problem of low survival rate

Active Publication Date: 2021-08-27
QINGDAO WOMEN & CHILDREN HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Early NB can regress spontaneously or simply surgical resection can achieve good prognosis, but children with advanced NB have a five-year survival rate of less than 40% despite intensive treatment measures

Method used

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  • Application of hsa_circ_0000284 in preparation of neuroblastoma prognostic preparation
  • Application of hsa_circ_0000284 in preparation of neuroblastoma prognostic preparation
  • Application of hsa_circ_0000284 in preparation of neuroblastoma prognostic preparation

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Experimental program
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Effect test

Embodiment 1

[0033] Preparation of hsa_circ_0000284 kit for the prognosis of patients with neuroblastoma (20 reactions)

[0034] 1. RNA stabilization solution 50ml

[0035] 2. Isopropanol 30ml

[0036] 3. Chloroform 20ml

[0037] 4. Trizol 30ml

[0038] 5. Enzyme-free water 20ml*2

[0039] 6. Ethanol 5ml

[0040] 7. gDNA Eraser 30μl

[0041] 8. 5×gDNA EraserBuffer 60μl

[0042] 9.RT Primer Mix 30μl

[0043] 10. 5×PrimeScript Buffer 2 (including dNTP Mixture) 90ml

[0044] 11. PrimeScript RT Enzyme Mix I (contains RNase inhibitor) 20μl

[0045] 12.TB Green Premix Ex Taq II 50μl

[0046] 13. 10μM hsa_circ_0000284 specific primer 30μl

[0047] Forward primer 5'-TATGTTGGTGGATCCTGTTCGGCA-3', SEQ ID NO:2;

[0048] Reverse primer 5'-TGGTGGGTAGACCAAGACTTGTGA-3', SEQ ID NO:3.

[0049] 14. 10μM GAPDH specific primer 30μl:

[0050] Forward primer 5'-TGCACCACCAACTGCTTAGC-3', SEQ ID NO:4;

[0051] Reverse primer 5'-GGCATGGACTGTGGTCATGAG-3', SEQ ID NO:5.

Embodiment 2

[0053] Detection of hsa_circ_0000284 in tumor tissues and paracancerous tissues of patients with neuroblastoma

[0054] 1. Sample pretreatment: collect the neuroblastoma tissue and paracancerous tissue to be tested respectively, store them in cryopreservation tubes filled with RNA stabilization solution, and put them in a -80°C refrigerator for later use.

[0055]2. Extraction of RNA: Take an appropriate amount of sample, add liquid nitrogen to the mortar after baking at 180°C for 6-8 hours to grind the sample, grind it to powder, add 1ml Trizol mortar sample to the mortar, and grind it into a liquid state Afterwards, transfer to the EP tube, add 200 μl / ml Trizol of chloroform to the EP, shake vigorously for 15-30 seconds, let stand on ice for 5 minutes, and centrifuge at 12,000 rpm at 4°C for 15 minutes; carefully take the uppermost aqueous phase and transfer to a new EP tube Add an equal volume of isopropanol, mix it upside down gently, let it stand on ice for 10 minutes; ce...

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Abstract

The invention discloses application of hsa_circ_0000284 in preparation of a neuroblastoma prognostic preparation, and relates to the technical field of molecular biology. The invention further discloses a nucleotide sequence of the hsa_circ_0000284, wherein the nucleotide sequence of the hsa_circ_0000284 is shown as SEQ ID NO:1, and the nucleotide sequences of the specific primers of the hsa_circ_0000284 are shown as SEQ ID NO:2 and SEQ ID NO:3. The survival curve analysis of 38 patients with neuroblastoma shows that the patients with high expression of hsa_circ_0000284 have a relatively poor postoperative survival rate (P is equal to 0.0107). According to the present invention, the expression level of the hsa_circ_0000284 is detected, such that the prognosis judgment is performed on the neuroblastoma patient so as to allow the hsa_circ_0000284 to be used as the important index of the clinical diagnosis evaluation of the neuroblastoma.

Description

technical field [0001] The present invention relates to the technical field of molecular biology, and more specifically relates to the application of hsa_circ_0000284 in preparing neuroblastoma prognostic preparations. Background technique [0002] Neuroblastoma (NB) is the most common extracranial solid tumor in children, originating from the primitive neuroepithelial cells of the embryonic neural crest, and can develop anywhere along the sympathetic nervous system, most commonly in the adrenal medulla. Children with NB account for 8-10% of the number of children with tumors, about 15% of the deaths of children with cancer, and about 50% of them have had tumor metastasis at the time of diagnosis. Early NB can regress spontaneously or simple surgical resection can achieve a good prognosis, but despite intensive treatment measures, the five-year survival rate of children with advanced NB is still less than 40%. To determine the prognosis of patients with neuroblastoma in ord...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/686C12N15/11
CPCC12Q1/6886C12Q1/686C12Q2600/178C12Q2600/158C12Q2563/107C12Q2545/114
Inventor 鹿洪亭尉嘉斌
Owner QINGDAO WOMEN & CHILDREN HOSPITAL
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