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Application method of long non-coding RNA CRYM-AS1

An RNACRYM-AS1, long-chain non-coding technology, applied in the field of preparation of prognostic reagents for glioma patients, can solve the problems of low survival rate, high recurrence rate and unclear boundary of surrounding brain tissue in glioma patients, and achieve far-reaching clinical results. Meaning and Promotional Effects

Active Publication Date: 2016-05-25
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, glioma grows infiltratively and has unclear borders with the surrounding brain tissue. Even the most advanced neurosurgical techniques are difficult to achieve pathological complete resection. Therefore, the recurrence rate of glioma is very high after surgery. The survival rate of patients is low, with an average survival time of 3-5 years

Method used

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  • Application method of long non-coding RNA CRYM-AS1
  • Application method of long non-coding RNA CRYM-AS1
  • Application method of long non-coding RNA CRYM-AS1

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Example 1 Preparation of LncRNACRYM-AS1 kit for the prognosis of glioma patients (50 reactions)

[0026] 1. RNA stabilization solution 50ml

[0027] 2. Isopropanol 100ml

[0028] 3. Chloroform 100ml

[0029] 4. Trizol 50ml

[0030] 5. Enzyme-free water 10ml

[0031] 6.1 μM random reverse transcription primer 50 μl

[0032] 7.5× reverse transcription buffer 200ml

[0033] 8. 10mM base triphosphate deoxynucleotides 100μl

[0034] 9.40U / μl RNase inhibitor 500μl

[0035] 10.200U / μl MMLV reverse transcriptase 50μl

[0036] 11. Premix ExTaq 50μl

[0037] 12. 10 μL ncRNACRYM-AS1 specific primer 30 μl

[0038] Forward primer 5'-CTGGAAGGCACGTAAGC-3',

[0039] Reverse primer 5'-GAGGCGGCGATGTATTG-3'.

[0040] 13. 10 μM U6snRNA specific primer 30 μl

[0041] The forward primer is 5'-ATTGGAACGATACAGAGAAGATT-3'

[0042] The reverse primer was 5'-GGAACGCTTCACGAATTTG-3'.

Embodiment 2

[0043] Example 2 Detection of LncRNACRYM-AS1 in Glioma Tissue

[0044] 1. Preservation of glioma tissue: collect the glioma tissue to be tested and store it in a cryopreservation tube filled with RNA stabilization solution, and put it in a -80°C refrigerator for later use.

[0045]2. Extraction of RNA in tissues: Take an appropriate amount of specimen, add liquid nitrogen to the mortar after baking at 180°C for 6-8 hours, grind the specimen, grind it to powder, add 1ml Trizol mortar specimen in the mortar, and grind it into a liquid After the shape, transfer it to a tube, add 200μl / ml Trizol in chloroform to the Tube, shake it by hand for 15-30s, place it on ice for 5 minutes, and centrifuge at 12000g at 4°C for 15 minutes; carefully take the upper aqueous phase into a new tube, add pre-cooled iso Propanol 0.5ml / ml Trizol, mix well, put in refrigerator at -20°C for 20min, centrifuge at 12000g at 4°C for 10min; discard the supernatant, add 1-2ml ethanol diluted with 75% DEPC wa...

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Abstract

The invention discloses a method for applying long non-coding RNA (Ribonucleic Acid LncRNA) CRYM-AS1, that is, a method for preparing a prognosis preparation for glioma patients, and particularly for preparing a kit for predicting prognosis of glioma patients by using a real-time fluorescent quantitation analysis method. Research shows that the expression of LncRNA CRYM-AS1 in glioma tissue is highlighted, the prognosis of glioma patients with high LncRNA CRYM-AS1 expression is poorer than the prognosis of glioma patients with low LncRNA CRYM-AS1 expression, therefore, if the expression of the LncRNA CRYM-AS1 is applied to prognosis prediction on the glioma patients, strong molecular biology basis can be made for prediction on prognosis of the glioma patients, and great clinical significance and popularization are achieved.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology, and in particular relates to an application method of LncRNACRYM-AS1 in preparing a prognostic reagent for glioma patients. Background technique [0002] Glioma originates from the ectoderm, accounting for about 40.49% of nervous system tumors. Its location and onset time are not limited. It can occur in any part of the central nervous system, and the age of onset ranges from infants to the elderly. Moreover, glioma grows infiltratively and has unclear borders with the surrounding brain tissue. Even the most advanced neurosurgical techniques are difficult to achieve pathological complete resection. Therefore, the recurrence rate of glioma is very high after surgery. The survival rate of patients is low, with an average survival time of 3-5 years. Therefore, judging the prognosis of glioma patients in order to choose the best treatment plan and significantly improve the survival rate of pat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6851C12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178C12Q2531/113C12Q2563/107C12Q2561/113
Inventor 武明花刘长红余志斌徐刚李桂源
Owner CENT SOUTH UNIV