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Application of incrnalnc_004208 and its detection reagents in the preparation of glioma prognosis reagents

A technology for detecting glioma and detection reagents, which is applied in the field of tumor molecular biology and can solve problems such as unexplained specific mechanisms

Inactive Publication Date: 2020-07-28
XIANGYA HOSPITAL CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The latest research shows that in addition to MGMT, the abnormality of various signaling pathways such as epidermal growth factor receptor (EGFR) in tumor cells is involved in the occurrence of TMZ resistance, but the specific mechanism has not yet been elucidated.

Method used

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  • Application of incrnalnc_004208 and its detection reagents in the preparation of glioma prognosis reagents
  • Application of incrnalnc_004208 and its detection reagents in the preparation of glioma prognosis reagents
  • Application of incrnalnc_004208 and its detection reagents in the preparation of glioma prognosis reagents

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Transcriptome sequencing technology and lncRNA sequencing technology analysis of TMZ sensitive / resistant cell expression profiles

[0037] Transcriptome sequencing technology and lncRNA sequencing technology (Illumina HiSeq TM 2000), selected brain glioma T98G / U-118MG cell line and TMZ-resistant T98G-R / U-118MG-R cell line for expression profiling analysis. Before screening, we first used Cuffmerge software to merge the transcripts spliced ​​from each sample, and removed transcripts with uncertain strand orientations to obtain the complete transcriptome information for this sequencing. The screening process is divided into the following 5 steps ( figure 1 A):

[0038] step1: Screening of the number of transcript exons: filter a large number of low-expression, low-confidence single-exon transcripts in the transcriptome splicing results, and select transcripts with exon numbers >= 2 (if there is no special requirement, the plant will Transcripts with exon numb...

Embodiment 2

[0049] Example 2 Screening and verification of differentially expressed lncRNAs in glioma TMZ-resistant cells

[0050] (1) Extraction of total cellular RNA (TRIzol method)

[0051] Cells were seeded in 6-well plates at 37°C, 5% CO 2 Incubate in the incubator. After the cells were completely attached to the wall, 1000 μl TRIzol was added to each well. After standing still until completely dissolved, transfer to a 1.5ml EP tube. Add 0.2ml of chloroform, shake vigorously for 15 seconds, and place at room temperature for 3 minutes. Centrifuge at 4°C and 12000rpm for 15min, and transfer the supernatant to a new EP tube. Add 0.5ml of isopropanol and let stand at room temperature for 10min. After centrifugation at 4°C and 12,000 rpm for 10 minutes, a gelatinous precipitate appeared at the bottom of the EP tube. Discard the supernatant, wash the RNA pellet with 1ml of 75℅ ethanol, and centrifuge at 4°C, 7500rpm for 5min, discard the supernatant again. After air-drying for 5 min...

Embodiment 3

[0066] Example 3 Correlation between patient survival and lncRNA expression

[0067] 104 patients with glioma were followed up by telephone, and they were asked in detail about the time of onset, grade, treatment, recurrence, other diseases, use of other drugs, recurrence and death time, etc. And registered the survival time and status. Survival analysis of the expression of LNC_004208 in brain glioma tissues and the survival time and status of patients found that the progression-free survival of patients with high expression ( Figure 4 A) and the average survival time was significantly shorter than that of patients with low expression or no expression ( Figure 4 B).

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Abstract

The invention discloses an application of IncRNA LNC_004208 and detection reagent thereof in preparing a glioma prognosis reagent. Through research on and utilization of IncRNA sequencing, the invention finds that IncRNA LNC_004208 has ultrahigh expression in a temozolomide-resisted glioma cell line; through telephone follow-up for 104 glioma patients, detailed inquiry for their first onset time,level, treatment condition, relapse situation, other diseases, other drugs, relapse time, death time, and the like, and registration for survival time and state, a result shows that average survival time and progression free survival of high expression patients are obviously shorter than those of low expression or no expression patients and are related to reactivity of temozolomide; thus, LNC_004208 is a molecular marker related to glioma prognosis; IncRNA LNC_004208 is high in expression; prognosis of patients is poor.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology, and specifically relates to the application of IncRNALNC_004208 and its detection reagent in the preparation of glioma prognosis reagents and corresponding reagents. Background technique [0002] Glioma is the most common malignant tumor of the central nervous system, accounting for about 80% of primary malignant brain tumors. According to the European Neuro-Oncology Association (EANO) statistics, the annual incidence of glioma in the world is about 6 / 100,000 . The treatment of glioma is a worldwide problem, especially for malignant glioblastoma. Even if the optimal treatment plan is adopted, the prognosis of patients is still very unsatisfactory, and the average survival period is only 13.3-15 months. Temozolomide (TMZ) is a new drug with better efficacy in the treatment of glioma that has emerged in the past two decades. It has been approved by the US FDA and recommended by the National ...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12Q1/6886
CPCC12Q1/6886C12Q2600/118C12Q2600/178
Inventor 颜元良徐志杰龚志成陈曦霍雷李学军曾双双钱龙王翔刘万里熊小明
Owner XIANGYA HOSPITAL CENT SOUTH UNIV
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