CN114480658B - Gene marker for glioma prognosis and application thereof - Google Patents

Gene marker for glioma prognosis and application thereof Download PDF

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CN114480658B
CN114480658B CN202210243318.7A CN202210243318A CN114480658B CN 114480658 B CN114480658 B CN 114480658B CN 202210243318 A CN202210243318 A CN 202210243318A CN 114480658 B CN114480658 B CN 114480658B
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ftl
fth1
ferritin
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CN114480658A (en
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杨飞城
王岩
覃艳
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First Affiliated Hospital of Army Medical University
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Abstract

The present invention provides a biomarker for detecting glioma prognosis or temozolomide resistance, the biomarker comprising a first marker for displaying ferritin light chain (FTL) expression level and a second marker for displaying ferritin heavy chain 1 (FTH 1) expression level. The invention also provides application of the kit in preparation of diagnostic reagents. The invention provides a very clear indication index for prognosis of glioma, and provides guiding reference for clinical prognosis prediction and treatment mode of patients successfully.

Description

Gene marker for glioma prognosis and application thereof
Technical Field
The invention relates to the technical field of molecular biology, in particular to a gene marker for glioma prognosis and application thereof.
Background
Iron death (ferroptosis) is a new pattern of cell death, a non-apoptotic pattern of iron ion dependent cell death. In 2003, the application of the small molecule medicine Erastin to the action mechanism of Ras gene mutant tumor cells is discovered for the first time by Dolma et al, and until 2012 Dixon reports that the cell death mode depends on iron ions and active oxygen to induce lipid peroxidation so as to cause regulatory cell necrosis, and is named ferroptosis or iron death. The cell death mode is obviously different from other forms such as apoptosis, necrosis, autophagy and the like in morphology, biology and gene level, can not be prevented by apoptosis or necrosis inhibitors, but the cell iron death can be inhibited by a small molecule iron chelator deferoxamine (ferrostatin-1) and an antioxidant (liproxstatin-1). Iron death is very significant in tumor therapy, and kidney cancer and leukemia are known to be very sensitive to iron death, and have a very close relationship in cerebral stroke and liver injury. The study demonstrated that the nature of iron death is a metabolic disorder of intracellular lipid peroxides, probably caused by the disruption of the GSH-GPX4 antioxidant system. Furthermore, the normal metabolism of ironAnd homeostasis may protect cells from iron death damage. Iron has two oxidation states, ferrous iron (Fe 2+ ) Or iron (Fe) 3+ ) The iron redox cycle may affect the sensitivity of the cell to iron death. Fe (Fe) 3+ Binds to Transferrin (TF) in serum and is then taken up by the cell through transferrin receptors (TFRC and TFR 2) on the cell membrane. Cytoplasmic iron is stored in the iron storage protein complex ferritin, which consists of ferritin light chain (FTL) and ferritin heavy chain 1 (FTH 1).
The induction of tumor iron death is a very promising anticancer way, but it is not clear how to induce glioma to generate iron death, whether iron death inducer can affect invasive growth of glioblastoma cells, and which iron death-related regulatory factors are involved in temozolomide resistance and other problems. Therefore, no report has been found in the prior art on the use of ferritin light chain (FTL) and ferritin heavy chain 1 (FTH 1) as prognostic indicators, in particular as prognostic indicators for gliomas, or as resistance indicators for related chemotherapeutic drugs, etc.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a technical scheme of taking FTH1 and FTL as biomarkers for predicting prognosis and temozolomide resistance of glioma patients through verifying the prognosis effect of the glioma patients after radiotherapy by utilizing the FTH1 and the FTL and researching the correlation between MGMT mRNA level and temozolomide resistance.
A biomarker for detecting glioma prognosis or temozolomide resistance, the biomarker comprising a first marker for displaying ferritin light chain (FTL) expression levels and a second marker for displaying ferritin heavy chain 1 (FTH 1) expression levels;
the first marker is selected from one of FTL-mRNA for translation into ferritin light chain, whole peptide fragment of ferritin light chain or a partial peptide fragment featuring the same;
the second marker is selected from one of FTH1-mRNA for translation into ferritin heavy chain 1, the entire peptide of ferritin heavy chain 1 or a partial peptide characteristic thereof.
In one embodiment according to the invention, the FTL-mRNA sequence is SEQ ID NO. 1
In one embodiment according to the invention, the sequence of all peptide stretches of the ferritin light chain is SEQ ID NO. 2.
In one embodiment according to the invention, the FTH1-mRNA sequence is SEQ ID NO:3.
In one embodiment according to the invention, the sequence of all peptide stretches of the ferritin heavy chain 1 is SEQ ID NO. 4.
The invention also provides application of the biomarker in preparation of a diagnostic reagent for detecting glioma prognosis or temozolomide drug resistance.
The invention further provides a diagnostic kit for glioma prognosis or temozolomide resistance comprising a first component for detecting a first marker as described above, and a second component for detecting a second marker as described above;
the first component and the second component are each independently selected from one or more of a PCR primer, a probe or an antibody.
In one embodiment according to the invention, the PCR primer, probe or antibody contains a fluorescent group.
The technical scheme of the invention has the following beneficial effects:
the marker provided by the invention can be used for predicting the prognosis of a glioma patient, can be used as a biomarker for detecting drug resistance of temozolomide, provides a very clear indication index for the prognosis of glioma, and successfully provides a guiding reference for the clinical prognosis prediction and treatment mode of the patient.
Drawings
FIG. 1 is a correlation profile of example 1 in a correlation analysis for FTH1 and FTL versus TMZ resistance, wherein,
FIG. 1A is a gene set enriched in GPX4, FTH1 and FTL, respectively, by GSEA in a Hallmark background;
FIG. 1B is a Pearson correlation graph of several iron death-related modulators;
FIG. 1C is a graph of a GSE database analysis to obtain a high expression FTH1 and FTL gene drug-resistant gene capable of enriching TMZ;
FIG. 1D is a Pearson correlation of MGMT with the listed iron death-related regulatory factors, wherein FTH1 and FTL are the top 2-phase genetic maps;
FIG. 1E is MGMT High With MGMT Low Enrichment of up-regulated genes in glioma patients with a profile associated with high expression of FTH1 and FTL;
FIG. 1F is a graph showing statistics of half-mortality and western blotting of temozolomide after transfection of glioblastoma cells with a control group sh-control, sh-FTH1 or sh-FTL;
FIG. 2 is a correlation graph of poor correlation analysis of FTH1 and FTL low expression and prognosis in glioma patients; wherein,
FIG. 2A is a graph of glioma classification versus FTH1 and FTL protein expression in a glioma-queuing chip;
FIG. 2B is a graph of Kaplan-Meier survival analysis of different protein expression patterns based on glioma cohort immunohistochemical data;
FIGS. 2C and 2D are graphs of Kaplan-Meier survival analysis results for different mRNA expression patterns in the TCGA_GBMLGG database, respectively;
FIG. 2E is a Kaplan-Meier survival analysis map of different mRNA expression patterns in the CGA_GBMLGG database;
FIG. 2F is a graph of mRNA expression patterns of FTH1 and FTL in different molecular subtypes of GBM in the TCGA_GBM database;
FIG. 2G is a diagram showing the expression of FTH1 and FTL in different molecular subtypes of glioblastoma in TCGA_GBM database.
FIG. 3 is a photograph of immunohistochemical detection for high and low expression of FTH1 and FTL.
Detailed Description
In order to make the technical problems, technical solutions and advantages to be solved more apparent, the following detailed description will be given with reference to the accompanying drawings and specific embodiments.
Experimental materials:
glioma samples: glioma tissue chip (HBrG 171Su 01) was purchased from Shanghai core Biotechnology Co., ltd (http:// www.superchip.com.cn/index. Html)
The clinical experiments related to the invention are approved by the medical ethics committee of southwest hospitals of the army university of army (original third army university of medical science).
The software used in the present invention is GraphPad Prism 8.
TCGA_GBMLGG and TCGA_GBM mRNA expression databases are downloaded from https:// xenabrowser. High and low expression of genes are defined by median values.
Data analysis was performed using DAVID Bioinformatics Resources and Gene Set Enrichment Assay (GSEA). t-test P values <0.05 were considered statistically significant. Patient survival and prognosis were assessed using Kaplan-Meier survival curve and log-rank statistics. The relationship between different genes or proteins was analyzed using Pearson rank correlation.
Cell lines and culture conditions
Cell line LN229, primary GBM cell line (GBM 1), construction of idh1 mutants and wild type GBM cells were cultured in DMEM (Gibco) medium; all media were supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin. All cells were exposed to 5% CO at 37℃ 2 Is cultured in a humidified incubator. LN229 cell line was purchased from ATCC (Manassas, va., USA). GBM1 cells are derived from an autonomously established human glioblastoma cell line. Cells were treated with TMZ (Selleck, # 123705) and observed for cell death. Mycoplasma contamination of all cells was periodically detected using a Mycoalert Plus (Lonza) and the Mycoplasma cells used in the experiment were monitored negative.
EXAMPLE 1 correlation of FTH1 and FTL with TMZ resistance
Since induction of iron death enhances TMZ inhibition of GBM cells, first, several key regulatory factors for iron death are analyzed, including GPX4, SLC7a11, FTH1, FTL, TF, TFRC, TFR2, NQO1, and ATL1. And then, through analysis of the TCGA-GBMLGG database, the correlation between FTH1 and FTL and GPX4 is found to be the largest. GSEA of Hallmark Gene set shows that FTH1 High For FTH1 Low 、FTL High To FTL (fiber to the air) Low 、GPX4 High For GPX4 Low Most of the gene sets enriched are overlapping (fig. 1A). Notably, these three groups of cells continue to be enriched for a gene set of oxidative stress pathways, which has been demonstrated to have important regulatory effects on iron death in cancer cells. GPX4, FTH1 (SEQ ID NO: 4) and FTL (SEQ ID NO: 2) were highly expressed to enrich for genes associated with epithelial-mesenchymal transition, wherein the FTH1 and FTL enrichment scores were higher than GPX4 (FIG. 1B). There is a significant difference in the molecular subtype of glioblastoma in terms of prognosis and therapeutic response, interestingly, high levels of FTH1, FTL and GPX4 expression are associated with the mesenchymal subtype of glioblastoma, and the enrichment scores of FTH1 and FTL are higher than GPX4. It is known that the resistance of gene MGMT and TMZ has direct correlation, and the correlation of the key iron death regulatory factor and MGMT is analyzed through TCGA-GBMLGG database, so that the correlation of iron death regulatory factor and temozolomide resistance can be indirectly known. The results showed that the transcriptome profile of MGMT expression was highly correlated with expression of FTH1 and FTL1 genes (fig. 1C). Then, select to be at MGMT High GBM to MGMT of (2) Low More than 1.4 times up-regulated genes in glioblastoma as a relevant gene set for temozolomide resistance. GSEA database analysis suggested that high expression of FTH1 and FTL could significantly enrich for these genes (fig. 1D). To further investigate whether FTH1 and FTL expression was directly associated with TMZ resistance, genes up-regulated by more than 6-fold compared to normal control cell lines in drug-resistant LN229 were screened as TMZ-resistant genes by analyzing the TMZ-resistant cell line from GSE113510 and its parental cell line. GSEA suggests that high expression of FTH1 and FTL could significantly enrich for these genes (fig. 1E). The same half-lethality of temozolomide to cells was examined by proliferation experiments and found that down-regulating the expression of FTH1 or FTL genes could promote sensitivity of glioblastoma cells to TMZ, confirming that FTH1 and FTL play a key role in temozolomide resistance (fig. 1F).
Example 2 FTH1 and FTL are related to progression and poor survival of GBM
By immunohistochemical analysis of 171 glioma samples (cowrt-171), expression of FTH1 and FTL was found to increase with increasing glioblastoma grade I to IV (fig. 2A). Kaplan-Meier survival analysis of tissue chip shows that it is compatible with FTH1 Low Or FTL Low FTH1 compared to patients of (a) High Or FTL High The prognosis of the patient was significantly worse (fig. 2B). Cases where FTH1 and FTL are simultaneously highly expressed (FTH 1) High /FTL High ) Other expression states of FTH1 and FTL, such as FTH1, with significantly shorter survival times High /FTL Low 、FTH1 Low /FTL High Or FTH1 Low /FTL Low (FIG. 2C). Briefly, FTH1 High /FTL High Prognosis for glioma patients is significantly better than other non-FTH 1 patients High /FTL High Glioma patients were poor (fig. 2D). The data for tcga_gbmlgg also consistently indicate that FTH1 or FTL high expression correlates with reduced overall survival and progression-free interphase events in glioma patients (fig. 2E), and FTH1 High /FTL High The survival time of glioma patients is obviously lower than that of non-FTH 1 High /FTL High Glioma patients. Also mentioned above, glioblastoma molecular typing is divided into four types: the mesenchymal subtypes, canonical, pre-neuromorphic and neuromorphic, were utilized with the tcga_gbm database, with the mesenchymal subtype being the subtype that is generally considered the worst prognosis. Further analysis, it was found that FTH1 and FTL expression was significantly higher in glioblastoma patients of the mesenchymal subtype than in the other three subtypes (fig. 2F). FTH1 High /FTL High The distribution of cases also showed that more than half (53%) of the cases were of the interstitial subtype (96 out of 181), but only 17.8% of cases were in non-FTH 1 High /FTL High In cases where the mesenchymal subtype was present (62 out of 348 cases), FTH1 and FTL expression was also higher for non-mesenchymal subtypes than for typical mesenchymal subtypes (fig. 2G). In summary, FTH1 and FTL, particularly in combination, can be used to predict poor prognosis for glioma patients.
Example 3 verification experiments by Gene expression and Western blotting
FTH1 and FTL are biomarkers, 1. The expression of FTH1 and FTL can be judged by pathologically taking the tumor area tissues of a patient, and carrying out gene level detection or protein level detection.
1. Detection of gene level:
the DNA content of FTH1 and FTL can be detected by second generation sequencing of tumor tissue of a patient, or the mRNA level of FTH1 and FTL can be detected by polymerase chain reaction PCR, wherein the FTL-mRNA sequence is SEQ ID NO. 1, and the FTH1-mRNA sequence is SEQ ID NO. 3.
If the expression level of the FTH1 and FTL genes is higher than that of normal, the prognosis of the patient is indicated to be poor. This can be demonstrated in fig. 2B-E, as shown in fig. 2A: the higher the grade of glioma patients with high FTH1 and FTL expression, the higher the malignancy. FIGS. 2B-E show that the higher the FTH1 and FTL expression, the shorter the overall survival of the patient, and the shorter the no-progress interval, and that FTH1 and FTL are prognostic indicators of glioma.
2. Detection from protein level:
through the detection of a conventional clinical pathognomonic immunohistochemical staining method or an immunoblotting method, if the FTH1 and the FTL are positively expressed or strongly positively expressed, the prognosis of the patient is indicated to be poor, and the FTH1 and the FTL are used as gene markers, can be used for prognosis judgment indexes and temozolomide drug resistance indexes of glioma patients, namely, a clinician can sample tumor tissues of the patient and then perform immunohistochemical detection, or second generation sequencing detection is carried out, if the expression of the FTH1 and the FTL proteins is found to be higher than the normal level, the patient can be indicated to be resistant to temozolomide, and the prognosis is poor. For example, FIG. 3 shows the immunohistochemical detection pictures of high expression and low expression of FTH1 and FTL, and if the detection of the FTH1 and FTL genes of a patient shows positive effect on the left picture, the prognosis is not good.
2.1 Western blotting Experimental procedure was as follows:
1) Collecting cells and tissues;
2) Cell/tissue lysis and protein quantification:
3) SDS-PAGE gel preparation
4) Protein loading and electrophoresis:
5) Transfer film
6) Blocked protein antigens
7) Antigen-antibody reaction
An antibody: anti-Ferritin Light Chain antibody (ab 69090 #abcam);
Anti-Ferritin Heavy Chain antibody (ab 65080# abcam)
8) And (5) developing.
2.2 immunohistochemical staining IHC Standard procedure
1) Slicing, baking at 60deg.C for 1 hr;
2) Dewaxing and rehydration
10min of dimethylbenzene, 5min of 100% ethanol, 5min of 95% ethanol, 5min of 90% ethanol, 5min of 85% ethanol, 5min of 80% ethanol, 5min of 75% ethanol, 5min of 60% ethanol, 5min of 50% ethanol, 5min of 30% ethanol, 1min of tap water and 1min of hydrogen peroxide;
3)3％H 2 O 2 washing with distilled water for 3 times at 37deg.C for 30min each for 3min;
4) Microwave repair
Immersing the slices in 0.01M EDTA buffer solution, heating the slices to boiling with the maximum fire power (98-100 ℃) in microwaves,
5) Naturally cooling the slice to room temperature, and washing with PBS for 3 times, each time for 5min;
6) Blocking, 5% BSA, and throwing away redundant liquid at room temperature for 20min;
7) Dripping primary antibody at 37 ℃ for 1h or at 4 ℃ overnight;
8) Washing with PBS for 3 times and 3min each time;
9) Dripping secondary antibody at 37deg.C for 15-30min;
10 PBS 3 times for 3min each;
11 Dripping SABC at 37deg.C for 30min;
12 PBS 3 times for 5min each;
13 1ml of distilled water is respectively dripped with the color-developing agent and evenly mixed;
14 After the DAB color developing agent is prepared, dripping the DAB color developing agent into slices, detecting the reaction time (about 5 min) under a mirror at room temperature;
15 Washing the water completely, and passing distilled water;
16 Hematoxylin counterstain for 2min, washing with tap water;
17 Dewatering
30% ethanol 3min,50% ethanol 3min,70% ethanol 3min,80% ethanol 3min,90% ethanol 3min,95% ethanol 3min,100% ethanol 3min, xylene 20min;
18 Gum sealing sheet, microscopy.
As shown in fig. 3, in fig. 3 a, a picture with high FTH1 expression (left) and a picture with low expression (right); in fig. 3B, i.e. the high-expression picture of FTL (left) and the low-expression picture of FTL (right), if the clinical test finds that the expression of FTL1 and FTL is positive as in the left panel, it indicates that the prognosis of the glioma patient is not good.
While the foregoing is directed to the preferred embodiments of the present invention, it will be appreciated by those skilled in the art that various modifications and adaptations can be made without departing from the principles of the present invention, and such modifications and adaptations are intended to be comprehended within the scope of the present invention.
FTL-mRNA sequence SEQ ID NO. 1
>NM_000146.4Homo sapiens ferritin light chain(FTL),mRNA
The sequence of the complete peptide stretch of the ferritin light chain SEQ ID NO. 2
>NP_000137.2ferritin light chain[Homo sapiens]
FTH1-mRNA sequence SEQ ID NO. 3
>NM_002032.3Homo sapiens ferritin heavy chain 1(FTH1),mRNA
The sequence of the complete peptide stretch of ferritin heavy chain 1 SEQ ID NO. 4
>NP_002023.2ferritin heavy chain[Homo sapiens]
Sequence listing
<110> first affiliated Hospital of the university of the force army of the Chinese people's liberation army
<120> a gene marker for glioma prognosis and use thereof
<141> 2022-03-11
<160> 4
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<400> 1
gcagttcggc ggtcccgcgg gtctgtctct tgcttcaaca gtgtttggac ggaacagatc 60
cggggactct cttccagcct ccgaccgccc tccgatttcc tctccgcttg caacctccgg 120
gaccatcttc tcggccatct cctgcttctg ggacctgcca gcaccgtttt tgtggttagc 180
tccttcttgc caaccaacca tgagctccca gattcgtcag aattattcca ccgacgtgga 240
ggcagccgtc aacagcctgg tcaatttgta cctgcaggcc tcctacacct acctctctct 300
gggcttctat ttcgaccgcg atgatgtggc tctggaaggc gtgagccact tcttccgcga 360
attggccgag gagaagcgcg agggctacga gcgtctcctg aagatgcaaa accagcgtgg 420
cggccgcgct ctcttccagg acatcaagaa gccagctgaa gatgagtggg gtaaaacccc 480
agacgccatg aaagctgcca tggccctgga gaaaaagctg aaccaggccc ttttggatct 540
tcatgccctg ggttctgccc gcacggaccc ccatctctgt gacttcctgg agactcactt 600
cctagatgag gaagtgaagc ttatcaagaa gatgggtgac cacctgacca acctccacag 660
gctgggtggc ccggaggctg ggctgggcga gtatctcttc gaaaggctca ctctcaagca 720
cgactaagag ccttctgagc ccagcgactt ctgaagggcc ccttgcaaag taatagggct 780
tctgcctaag cctctccctc cagccaatag gcagctttct taactatcct aacaagcctt 840
ggaccaaatg gaaataaagc tttttgatgc a 871
<210> 2
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<212> PRT
<213> Human (Human)
<400> 2
Met Ser Ser Gln Ile Arg Gln Asn Tyr Ser Thr Asp Val Glu Ala Ala
1 5 10 15
Val Asn Ser Leu Val Asn Leu Tyr Leu Gln Ala Ser Tyr Thr Tyr Leu
20 25 30
Ser Leu Gly Phe Tyr Phe Asp Arg Asp Asp Val Ala Leu Glu Gly Val
35 40 45
Ser His Phe Phe Arg Glu Leu Ala Glu Glu Lys Arg Glu Gly Tyr Glu
50 55 60
Arg Leu Leu Lys Met Gln Asn Gln Arg Gly Gly Arg Ala Leu Phe Gln
65 70 75 80
Asp Ile Lys Lys Pro Ala Glu Asp Glu Trp Gly Lys Thr Pro Asp Ala
85 90 95
Met Lys Ala Ala Met Ala Leu Glu Lys Lys Leu Asn Gln Ala Leu Leu
100 105 110
Asp Leu His Ala Leu Gly Ser Ala Arg Thr Asp Pro His Leu Cys Asp
115 120 125
Phe Leu Glu Thr His Phe Leu Asp Glu Glu Val Lys Leu Ile Lys Lys
130 135 140
Met Gly Asp His Leu Thr Asn Leu His Arg Leu Gly Gly Pro Glu Ala
145 150 155 160
Gly Leu Gly Glu Tyr Leu Phe Glu Arg Leu Thr Leu Lys His Asp
165 170 175
<210> 3
<211> 1203
<212> DNA
<213> Human (Human)
<400> 3
gccagacgtt cttcgccgag agtcgtcggg gtttcctgct tcaacagtgc ttggacggaa 60
cccggcgctc gttccccacc ccggccggcc gcccatagcc agccctccgt cacctcttca 120
ccgcaccctc ggactgcccc aaggcccccg ccgccgctcc agcgccgcgc agccaccgcc 180
gccgccgccg cctctcctta gtcgccgcca tgacgaccgc gtccacctcg caggtgcgcc 240
agaactacca ccaggactca gaggccgcca tcaaccgcca gatcaacctg gagctctacg 300
cctcctacgt ttacctgtcc atgtcttact actttgaccg cgatgatgtg gctttgaaga 360
actttgccaa atactttctt caccaatctc atgaggagag ggaacatgct gagaaactga 420
tgaagctgca gaaccaacga ggtggccgaa tcttccttca ggatatcaag aaaccagact 480
gtgatgactg ggagagcggg ctgaatgcaa tggagtgtgc attacatttg gaaaaaaatg 540
tgaatcagtc actactggaa ctgcacaaac tggccactga caaaaatgac ccccatttgt 600
gtgacttcat tgagacacat tacctgaatg agcaggtgaa agccatcaaa gaattgggtg 660
accacgtgac caacttgcgc aagatgggag cgcccgaatc tggcttggcg gaatatctct 720
ttgacaagca caccctggga gacagtgata atgaaagcta agcctcgggc taatttcccc 780
atagccgtgg ggtgacttcc ctggtcacca aggcagtgca tgcatgttgg ggtttccttt 840
accttttcta taagttgtac caaaacatcc acttaagttc tttgatttgt accattcctt 900
caaataaaga aatttggtac ccaggtgttg tctttgaggt cttgggatga atcagaaatc 960
tatccaggct atcttccaga ttccttaagt gccgttgttc agttctaatc acactaatca 1020
aaaagaaacg agtatttgta tttattaaac tcattagttt gggcagtata ctaaggtgtg 1080
gctgtcttgg attcagatag aactaagggt tcccgactct gaatccagag tctgagttaa 1140
atgtttccaa tggttcagtc tagctttcac agtttttatg aataaaaggc attaaaggct 1200
gaa 1203
<210> 4
<211> 183
<212> PRT
<213> Human (Human)
<400> 4
Met Thr Thr Ala Ser Thr Ser Gln Val Arg Gln Asn Tyr His Gln Asp
1 5 10 15
Ser Glu Ala Ala Ile Asn Arg Gln Ile Asn Leu Glu Leu Tyr Ala Ser
20 25 30
Tyr Val Tyr Leu Ser Met Ser Tyr Tyr Phe Asp Arg Asp Asp Val Ala
35 40 45
Leu Lys Asn Phe Ala Lys Tyr Phe Leu His Gln Ser His Glu Glu Arg
50 55 60
Glu His Ala Glu Lys Leu Met Lys Leu Gln Asn Gln Arg Gly Gly Arg
65 70 75 80
Ile Phe Leu Gln Asp Ile Lys Lys Pro Asp Cys Asp Asp Trp Glu Ser
85 90 95
Gly Leu Asn Ala Met Glu Cys Ala Leu His Leu Glu Lys Asn Val Asn
100 105 110
Gln Ser Leu Leu Glu Leu His Lys Leu Ala Thr Asp Lys Asn Asp Pro
115 120 125
His Leu Cys Asp Phe Ile Glu Thr His Tyr Leu Asn Glu Gln Val Lys
130 135 140
Ala Ile Lys Glu Leu Gly Asp His Val Thr Asn Leu Arg Lys Met Gly
145 150 155 160
Ala Pro Glu Ser Gly Leu Ala Glu Tyr Leu Phe Asp Lys His Thr Leu
165 170 175
Gly Asp Ser Asp Asn Glu Ser
180

Claims (5)

1. Use of a first marker for displaying the expression level of ferritin light chain (FTL) and a second marker for displaying the expression level of ferritin heavy chain 1 (FTH 1) in the preparation of a diagnostic reagent for detecting glioblastoma prognosis or temozolomide resistance;
the first marker is selected from FTL-mRNA for translation to ferritin light chain, all peptide fragments of ferritin light chain; the sequence of all peptide segments of the ferritin light chain is SEQ ID NO. 2;
the second marker is selected from FTH1-mRNA for translation into ferritin heavy chain 1, and all peptide fragments of ferritin heavy chain 1, and the sequence of all peptide fragments of ferritin heavy chain 1 is SEQ ID NO 4.
2. The use of claim 1, wherein said FTL-mRNA sequence is SEQ ID No. 1.
3. The biomarker of claim 1, wherein the FTH1-mRNA sequence is SEQ ID No. 3.
4. Use of a first component for detecting a first marker exhibiting the expression level of a ferritin light chain (FTL) having the sequence of SEQ ID NO 2 and a second component for detecting a second marker exhibiting the expression level of a ferritin heavy chain 1 (FTH 1) having the sequence of SEQ ID NO 4 for the preparation of a diagnostic kit for glioblastoma prognosis or temozolomide resistance;
the first component and the second component are each independently selected from one or more of a PCR primer, a probe or an antibody.
5. The use of claim 4, wherein the PCR primer, probe or antibody contains a fluorescent group.
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