CN113278702A - Application of PSMC2 gene detection primer in preparation of glioblastoma multiforme auxiliary diagnosis and prognosis evaluation kit - Google Patents
Application of PSMC2 gene detection primer in preparation of glioblastoma multiforme auxiliary diagnosis and prognosis evaluation kit Download PDFInfo
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Abstract
The PSMC2 gene detection primer is applied to preparation of a glioblastoma multiforme auxiliary diagnosis and prognosis evaluation kit, and belongs to the field of biomedical specialty. The detection primer based on the PSMC2 gene is used for preparing a glioblastoma multiforme auxiliary diagnosis and prognosis evaluation kit, the malignancy degree of a glioma patient is diagnosed by detecting the expression condition of the transcription level of the PSMC2 gene, the prognosis condition of the glioma patient is estimated, the prognosis of the patient is judged more accurately and objectively, and the radiotherapy and chemotherapy sensitivity of the patient can be evaluated. The glioblastoma is evaluated to be more sensitive and specific by using a mode of detecting the transcriptional level expression of the PSMC2 gene of a patient with the glioma by using a detection primer.
Description
Technical Field
The invention belongs to the field of biomedical specialty, and particularly relates to an application of a PSMC2 gene detection primer in preparation of a glioblastoma multiforme auxiliary diagnosis and prognosis evaluation kit.
Background
Gliomas are the most common intracranial malignancies, accounting for 74.6% of those in the central nervous system. Existing traditional treatments, such as surgical resection with the assistance of radiation and chemotherapy, do not better improve the overall survival of the patient. Glioblastoma (GBM) is the most common and malignant glioma, with a median survival of only 15 months. Exploring molecules that play a key role in glioblastoma malignant progression would help improve patient survival.
The 26S proteasome is a multicatalytic protease complex with a highly ordered structure consisting of 2 complexes (20S core and 19S regulatory particle). The 20S core consists of 4 loops of 28 different subunits; 2 loops consist of 7 alpha subunits, while 2 loops consist of 7 beta subunits. The 19S regulatory particle consists of bases comprising 6 ATPase subunits and 2 non-ATPase subunits and a cap containing up to 10 non-ATPase subunits. Proteasomes are distributed in high concentrations throughout eukaryotic cells and cleave peptide fragments in an ATP/ubiquitin dependent process in a non-lysosomal pathway. The basic function of the modified proteasome is to process MHC-class I peptides. PSMC2 is an essential component of 19S regulatory particles of 26S proteasome, can be combined with ATP and nucleotide, has the functions of hydrolase and nucleoside triphosphatase, plays a key role in selectively degrading protein in cells, and participates in regulating cell differentiation and proliferation, apoptosis, energy metabolism, signal transduction and other biological activities. Research shows that the PSMC2 and 26S proteasome are closely related to the occurrence and development of various tumors, including breast cancer, hepatocellular carcinoma, esophageal cancer, pancreatic cancer and the like, and the role of PSMC2 in glioma is unknown. Therefore, the role of PSMC2 in glioma needs to be explored urgently and markers are developed to provide assistance for diagnosis and treatment of glioma.
At present, no kit for the PSMC2 gene in glioblastoma has been reported. The invention describes the functional role of the PSMC2 gene in glioblastoma, and the PSMC2 gene is prepared into a kit for evaluating poor prognosis and the effect of radiotherapy and chemotherapy.
Disclosure of Invention
In view of the defects in the prior art, the invention mainly provides the application of the detection primer of the PSMC2 gene in the preparation of a glioblastoma multiforme auxiliary diagnosis and prognosis evaluation kit, finds that the transcription expression level of the PSMC2 gene is increased along with the increase of the tumor grade, and can diagnose the malignancy degree of a glioma patient by detecting the expression condition of the transcription level of the PSMC2 gene. Meanwhile, the prognosis condition and the effect of radiotherapy and chemotherapy of the patient can be evaluated according to the expression condition of the PSMC2 gene.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows.
A PSMC-aided diagnosis and prognosis evaluation kit based on a PSMC2 gene is used for detecting the PSMC gene level of PSMC2, and comprises a primer for detecting the PSMC2 gene level.
The primers for detecting the PSMC2 gene level comprise a PCR primer pair for amplifying the PSMC2 gene, wherein the PCR primer pair for amplifying the PSMC2 gene comprises a forward primer sequence 5'-AGACACTCCAGAGTGAACAGCC-3', a reverse primer sequence 5'-CTAAGGTCCACCACAAACTTGGC-3', a forward primer sequence 5'-GAGCACTTACTCTAGGCAGATCA-3', a reverse primer sequence 5'-GTACACCTGGCAACCTGTAAAG-3' or a forward primer sequence 5'-GATTCGGAGGACCCAAAATACAT-3' and a reverse primer sequence 5'-CCACTCTCATCCCTTCTTCAATG-3';
the primer for detecting the PSMC2 gene level further comprises a PCR primer pair for amplifying the housekeeping gene 18s, wherein the PCR primer pair for amplifying the housekeeping gene 18s is as follows: the forward primer sequence was 5'-ACCCGTTGAACCCCATTCGTGA-3' and the reverse primer sequence was 5'-GCCTCACTAAACCATCCAATCGG-3'.
The kit also comprises a SYBR Green polymerase chain reaction system, wherein the SYBR Green polymerase chain reaction system comprises a PCR buffer solution, dNTPs, SYBR Green fluorescent dye, enzyme-free water and a fluorescent quantitative sample adding plate.
The kit also comprises an RNA extraction reagent, wherein the RNA extraction reagent comprises Trizol, chloroform, isopropanol, 75% ethanol and RNase free water.
The kit also comprises a system for reverse transcribing the mRNA into the cDNA, wherein the system for reverse transcribing the mRNA into the cDNA adopts a reverse transcription complex reagent 5X RT mastermix, the reverse transcription complex reagent 5X RT mastermix comprises PrimeScriptRTase, RNase Inhibitor, Random 6 mers, Oligo dT Primer, dNTPmix and reaction Buffer, and the fluorescent quantitative PCR reaction system comprises a fluorescent quantitative PCR reaction enzyme SYBR Premix Ex TaqTM.
A glioblastoma multiforme auxiliary diagnosis and prognosis evaluation kit based on a PSMC2 gene and a using method thereof comprise the following steps.
a) And (3) treating and grinding the obtained fresh tissue by liquid nitrogen, and then extracting RNA.
b) The extracted RNA was reverse transcribed into the corresponding cDNA.
c) The reverse transcribed cDNA was subjected to fluorescent quantitative PCR amplification of PSMC2 and 18s genes.
d) Taking 18s as an internal reference, recording the Ct value of each reaction, and expressing the detection result as-delta Ct, wherein the delta Ct is CtPSMC2-Ct18sIn 2 of-ΔCtAs an index for evaluating the expression level of PSMC2 gene.
Wherein the Ct value is the cycle number of the fluorescence signal in each reaction tube reaching a set threshold value, CtPSMC2Is the cycle number, Ct, required for the amplification of PSMC2 expression in the reaction tube18sThe number of cycles required for expression of 18s to be amplified in the reaction tube.
The PSMC2 gene detection primer is applied to preparation of a glioblastoma multiforme auxiliary diagnosis and prognosis evaluation kit. The kit is used for detecting the gene level of PSMC2 of glioblastoma multiforme, and comprises a primer for detecting the gene level of PSMC 2.
The kit is used for grading the malignancy degree of the glioblastoma.
The kit is used for evaluating the prognosis condition and the effect of radiotherapy and chemotherapy of a patient with glioblastoma.
Compared with the prior art, the invention has the following advantages and beneficial effects.
(1) The invention discloses that the PSMC2 gene is related to glioma disease progression for the first time, and the PSMC2 gene is expected to provide a new idea for researching the molecular mechanism of glioma disease progression.
(2) The transcriptional expression level of the PSMC2 gene is increased along with the increase of the glioma grade, and the malignancy degree of a glioma patient can be diagnosed by detecting the transcriptional expression condition of the PSMC2 gene, so that a glioblastoma auxiliary diagnosis kit containing a PSMC2 gene detection primer can be prepared, and the glioblastoma can be evaluated more sensitively and more specifically by using a mode of detecting the transcriptional expression of the PSMC2 gene of the glioma patient by using the detection primer.
(3) The invention firstly proves that the expression level of the PSMC2 gene in glioma and even glioblastoma is directly related to the prognosis of patients, and the survival time of the patients with high PSMC2 is obviously reduced compared with the patients with low PSMC2 expression. The expression condition of the transcription level of PSMC2 is detected by the detection primer, and the prognosis condition of a patient with glioma can be estimated, so that the glioblastoma prognosis evaluation kit containing the PSMC2 gene primer is prepared, the prognosis of the patient can be more accurately and objectively judged, and the chemoradiotherapy sensitivity of the patient can be evaluated.
Drawings
FIG. 1 is a graph showing the correlation of PSMC2 gene expression level with malignancy phenotype in tumor grade, pathological typing and TCGA typing.
FIG. 2 is a graph of PSMC2 gene expression level versus survival in glioma patients.
FIG. 3 is a graph showing the relationship between PSMC2 gene expression level and GBM patient survival in chemotherapy.
Detailed Description
The invention is explained in more detail below with reference to specific embodiments and the drawing. The following examples are merely illustrative of the present invention and should not be construed as limiting thereof. The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental reagents and materials involved are conventional biochemical reagents and materials unless otherwise specified.
Example 1: the PSMC2 gene has significant difference in database and is a marker for malignant progression of glioma.
The research mainly comprises three Glioma database platforms, namely a Chinese Glioma Gene Atlas (CGGA), a tumor gene Atlas plan (TCGA) and GSE 16011. We collected expression profile data and clinical information data of all glioma patients in three databases, and used the R language "ggplot 2" package to draw a map of differences in PSMC2 expression among patients of different grades, different pathological types, and different TCGA types, as shown in fig. 1, in the cggarseq, TCGA RNAseq, and GSE16011 databases, the PSMC2 gene transcript level significantly increased with the increase in glioma grade; glioblastomas have the highest expression of PSMC2 according to histopathological classification; furthermore, PSMC2 was significantly higher in patients with interstitial glioma than in patients with classical and pre-neuronal types. The above results indicate that there is a close correlation between PSMC2 expression and glioma malignant phenotype.
Example 2: cox regression analysis was performed on glioma patients based on CGGA RNAseq, TCGA RNAseq and GSE16011 databases. The grade, IDH1 mutation status and expression level of PSMC2 were included in a Cox multi-factor regression analysis model and these factors were analyzed for relationship to glioma prognosis. The analysis results are shown in tables 1, 2 and 3, and show that the expression level of PSMC2 has the value of independently judging prognosis.
Example 3: survival time is used as an abscissa and survival rate is used as an ordinate in databases of CGGA RNAseq, TCGA RNAseq and GSE16011, and a PSMC2 expression level graph is drawn.
In the CGGA RNAseq database, survival curves were first drawn for glioma patients, ordered from small to large according to the mRNA expression levels of PSMC2, with the first 155 cases being the low PSMC2 expression group and the last 155 cases being the high PSMC2 expression group. FIG. 2A is a survival curve based on the level of mRNA expression of PSMC2 gene in CGGA RNAseq database. P <0.0001 by longrank test analysis, namely the prognosis is statistically significant. In addition, the relation between the expression level of PSMC2 and the prognosis is also explored in different grades of glioma patients. In FIG. 2, B is a survival curve of a glioma grade II patient in the CGGA RNAseq database based on the expression level of mRNA of the PSMC2 gene. P =0.0012 by longrank test analysis, i.e. the prognosis is statistically significant. FIG. 2C is a survival curve of a patient of glioma grade III based on the level of mRNA expression of the PSMC2 gene in the CGGA RNAseq database. P =0.0353 by longrank test analysis, i.e. the prognosis is statistically significant. FIG. 2, panel D is a survival curve of patients with grade IV glioma (i.e., glioblastoma, GBM) in the CGGA RNAseq database based on the mRNA level of PSMC2 gene. P =0.0422 was analyzed by longrank test, i.e. the prognosis was statistically significant.
In the tcgarseq database, survival curves were first drawn for glioma patients, based on the mRNA expression levels of PSMC2, ranked from small to large, with the first 316 cases being low PSMC2 expression groups and the last 315 cases being high PSMC2 expression groups. FIG. 2, panel E is the survival curve chart of TCGARNAseq database based on the high and low mRNA expression level of PSMC2 gene. P <0.0001 by longrank test analysis, namely the prognosis is statistically significant. In addition, the relation between the expression level of PSMC2 and the prognosis is also explored in different grades of glioma patients. FIG. 2F is a graph showing the survival of glioma grade II patients in the TCGARNAseq database based on the high and low mRNA expression level of PSMC2 gene. P =0.1824 by longrank test analysis, i.e. the prognosis has marginal statistical significance. FIG. 2, panel G is a graph of glioma grade III patient survival based on the high or low mRNA expression of PSMC2 gene in TCGARNAseq database. P =0.0004 by longrank test analysis, i.e. the prognosis is statistically significant. FIG. 2, panel H is a graph of survival of patients with glioma grade IV (i.e., glioblastoma, GBM) based on the high and low mRNA expression of PSMC2 gene in TCGARNAseq database. P =0.0482 was analyzed by longrank test, i.e. the prognosis was statistically significant.
In the GSE16011 database, survival curves were first drawn for glioma patients, with the mRNA expression levels of PSMC2 ranked from small to large, the first 132 cases being low PSMC2 expression sets, and the last 132 cases being high PSMC2 expression sets. FIG. 2 is a graph showing the survival rate of the GSE16011 database based on the level of mRNA expression of PSMC2 gene. P <0.0001 by longrank test analysis, namely the prognosis is statistically significant. In addition, the relation between the expression level of PSMC2 and the prognosis is also explored in different grades of glioma patients. In FIG. 2, J is the survival curve of patients with glioma grade II in GSE16011 database based on the high or low mRNA expression level of PSMC2 gene. P =0.0677 by longrank test analysis, i.e. the prognosis has marginal statistical significance. FIG. 2, K is a graph showing the survival rate of patients with grade III glioma in GSE16011 database based on the high or low mRNA expression level of PSMC2 gene. P =0.0130 by longrank test analysis, i.e. the prognosis is statistically significant. In FIG. 2, L is the survival graph of patients with GSE16011 database glioma grade IV (i.e., glioblastoma, GBM) based on the high and low mRNA expression of PSMC2 gene. P =0.0024 by longrank test analysis, i.e. the prognosis is statistically significant.
In conclusion, the expression level of the PSMC2 gene can reflect the prognosis of glioma patients, i.e. glioma patients with high PSMC2 expression often have short survival time.
Example 4: and (3) aiming at the radiotherapy condition of the patient, drawing a survival curve according to the expression level of PSMC2 by taking the survival time as an abscissa and the survival rate as an ordinate in the CGGA RNAseq database.
In a CGGA RNAseq database, aiming at whether GBM patients receive radiotherapy and PSMC2 expression conditions, the GBM patients are divided into four groups of radiotherapy + PSMC2 low expression, radiotherapy + PSMC2 high expression, non-radiotherapy + PSMC2 low expression and non-radiotherapy + PSMC2 high expression, survival curves are drawn according to the four groups of patients, and A in a graph in a figure 3 is a survival curve chart of the GBM patients irradiated by the CGGA RNAseq database based on the mRNA expression of PSMC2 gene. Among patients receiving radiotherapy, the survival time of patients with the PSMC2 low expression group is higher than that of patients with the PSMC2 high expression group, and the longrank test analysis P = 0.0496 has statistical significance. FIG. 3, panel B is a graph of GBM patient survival without the CGGA RNAseq database receiving radiation therapy based on PSMC2 gene mRNA expression. In patients who did not receive radiotherapy, the survival time of patients with PSMC2 in the low expression group was not significantly different from that of patients with PSMC2 in the high expression group, and the longrank test analysis P = 0.1218 had no statistical significance. Taken together, the high expression of the PSMC2 gene resulted in GBM patients' resistance to radiation therapy.
Example 5: survival curves were plotted against patient chemotherapy in the CGGA RNAseq database according to the level of PSMC2 expression.
In the CGGA RNAseq database, aiming at whether GBM patients receive chemotherapy and PSMC2 expression conditions, the GBM patients are divided into four groups of chemotherapy + PSMC2 low expression, chemotherapy + PSMC2 high expression, non-chemotherapy + PSMC2 low expression and non-chemotherapy + PSMC2 high expression, survival curves are drawn according to the four groups of patients, and C in the figure 3 is a PSMC2 gene mRNA expression high-low survival curve graph of the GBM patients receiving chemotherapy in the CGGA RNAseq database. Among patients receiving chemotherapy, patients with low expression PSMC2 survived longer than those with high expression PSMC2, and longrank test analysis P = 0.0625 had marginal statistical significance. FIG. 3D is a graph of GBM patient survival without the CGGA RNAseq database based on the level of mRNA expression of PSMC2 gene. In patients who did not receive chemotherapy, the survival time of patients in the low expression PSMC2 group was not significantly different from that in the high expression PSMC2 group, and the longrank test analysis P = 0.3781 had no statistical significance. Taken together, the high expression of the PSMC2 gene resulted in GBM patients' resistance to chemotherapy.
Example 6: and (3) preparation of a glioblastoma auxiliary diagnostic kit based on a PSMC2 gene detection primer.
The glioblastoma multiforme auxiliary diagnosis and prognosis evaluation kit prepared by the PSMC2 gene level detection primer comprises a PCR primer pair for amplifying the PSMC2 gene, a PCR primer pair for amplifying housekeeping gene 18s, a SYBR Green polymerase chain reaction system, an RNA extraction reagent, a system for reverse transcription of mRNA into cDNA and a fluorescent quantitative PCR reaction system.
The PCR primer pair for amplifying the PSMC2 gene comprises the following components:
the forward primer sequence is 5'-AGACACTCCAGAGTGAACAGCC-3'
The reverse primer sequence is 5'-CTAAGGTCCACCACAAACTTGGC-3';
the PCR primer pair for amplifying housekeeping gene 18s is as follows:
the forward primer sequence is 5'-ACCCGTTGAACCCCATTCGTGA-3'
The reverse primer sequence is 5'-GCCTCACTAAACCATCCAATCGG-3'.
The SYBR Green polymerase chain reaction system comprises PCR buffer solution, dNTPs, SYBR Green fluorescent dye, RNase free water and a fluorescent quantitative adding template.
The RNA extraction reagent comprises Trizol, chloroform, isopropanol, 75% ethanol and RNase-free water.
The system for reverse transcription into cDNA adopts a reverse transcription complex reagent 5X RT master mix, and the reverse transcription complex reagent 5X RT master mix comprises PrimeScriptRTase, RNaseInhibitor, Random 6 mers, Oligo dT Primer, dNTP mix and reaction Buffer. Here, 5X RT master mix used a complex reagent of Takara company under the trade name RR 036Q.
The fluorescent quantitative PCR reaction system comprises a fluorescent quantitative PCR reaction enzyme SYBR Premix Ex TaqTM. As the PCR reaction enzyme, there is used a PCR reaction enzyme having a product number of RR820A from Takara.
Example 7: an application method of a glioblastoma multiforme auxiliary diagnosis and prognosis evaluation kit prepared by PSMC2 gene level detection primers.
The detection process comprises the following specific steps.
1) Grinding fresh tissue to be detected under the action of liquid nitrogen, adding 1mL of Trizol into the broken tissue, repeatedly blowing and beating by using a 1mL pipette gun, incubating at room temperature for 5min, and fully separating the nucleoprotein compound.
2) 200ul of chloroform was added to each tube, the tubes were inverted 10 times, left at room temperature for 3min, centrifuged at 12000rpm at 4 ℃ for 15min, and the upper aqueous phase was transferred to a new EP tube (ca. 500 ul).
3) Adding isopropanol (about 500ul, precooled at 4 deg.C) with the same volume as the supernatant, mixing by inversion, standing at room temperature for 10-20min (average 15min), centrifuging at 12000rpm at 4 deg.C for 10min, discarding the supernatant to obtain a little white precipitate.
4) Adding 1ml of pre-cooled 75% ethanol (RNase-free water diluted absolute ethanol), slowly adding along the tube wall, shaking up and down gently, and centrifuging at 7500rpm at 4 deg.C for 5 min.
5) Removing supernatant, drying at room temperature for 1-2min, adding RNase-free water 50-100ul, and blowing gently with pipette if necessary.
6) The concentration and purity of RNA were determined by Nanodrop. The ratio of OD260/OD280 is 1.80-2.0, which indicates that the purity of RNA meets the experimental requirements.
7) And (4) a reverse transcription system.
1ug RNA volume: x ul.
5X PrimeScript RT Master Mix: 4ul。
RNase free water: 16-Xul.
The total system is as follows: 20 ul.
The reverse transcription conditions were: 15min at 37 ℃, 5S at 85 ℃ and 1min at 4 ℃.
Wherein, the volume X of 1ug RNA is calculated according to the RNA concentration, 1 microgram of RNA is needed, after the RNA is extracted through the steps 1-6, a concentration is obtained, and then the volume of the RNA solution is calculated according to the concentration, and the volume of the X cannot exceed 16ul at most.
8) Fluorescent quantitative PCR sample adding: the fluorescence quantification plate was placed on ice, 3 replicate wells were made for each sample, and the cDNA (diluted 4-5 times and 2. mu.L added after dilution) was added.
The fluorescent quantitative PCR system is provided.
SYBR Premix Ex TaqTM: 10ul。
The upstream primer (10 umol) is 1 ul.
The downstream primer (10 umol) is 1 ul.
cDNA template: 2 ul.
RNase-free water: 6 ul.
The total system is as follows: 20 ul.
9) After the sample adding is finished, the fluorescent quantitative plate film is covered with the sealing plate carefully by using a PE glove, and the fluorescent quantitative plate film is centrifuged to avoid the generation of bubbles.
10) Procedure for setting up fluorescent quantitative PCR reactions: the 40 cycles + dissolution curves were performed at 95 ℃ for 10min, 95 ℃ for 5S, 60 ℃ for 10S, and 72 ℃ for 10S.
11) And (3) analyzing experimental data: the Ct value for each reaction was recorded with 18s as internal reference, the Ct value being the number of cycles that the fluorescence signal in each reaction tube went through to reach the set threshold. Δ Ct ═ CtPSMC2-Ct18The smaller the Δ Ct, the higher the expression, 2, indicating the higher the initial copy number-ΔCtThe PSMC2 expression level was evaluated as an index.
Various other modifications and changes may be made by those skilled in the art based on the above-described technical solutions and concepts, and all such modifications and changes should fall within the scope of the claims of the present invention.
Claims (9)
- The application of the detection primer of the PSMC2 gene in preparing a glioblastoma multiforme auxiliary diagnosis and prognosis evaluation kit is characterized in that the kit is used for detecting the PSMC2 gene level of glioblastoma multiforme, and the kit contains the primer for detecting the PSMC2 gene level.
- 2. The use of claim 1, wherein the primers comprise a PCR primer pair for amplifying PSMC2 gene and a PCR primer pair for amplifying housekeeping gene 18 s;the PCR primer pair for amplifying the PSMC2 gene is a forward primer sequence 5'-AGACACTCCAGAGTGAACAGCC-3', a reverse primer sequence 5'-CTAAGGTCCACCACAAACTTGGC-3', a forward primer sequence 5'-GAGCACTTACTCTAGGCAGATCA-3', a reverse primer sequence 5'-GTACACCTGGCAACCTGTAAAG-3' or a forward primer sequence 5'-GATTCGGAGGACCCAAAATACAT-3' and a reverse primer sequence 5'-CCACTCTCATCCCTTCTTCAATG-3';the PCR primer pair for amplifying housekeeping gene 18s is a forward primer sequence 5'-ACCCGTTGAACCCCATTCGTGA-3' and a reverse primer sequence 5'-GCCTCACTAAACCATCCAATCGG-3'.
- 3. The use of claim 1, wherein the kit further comprises a SYBR Green polymerase chain reaction system comprising PCR buffer, dNTPs, SYBR Green fluorescent dye, enzyme-free water and a fluorescent quantitation plate.
- 4. The use of claim 1, wherein the kit further comprises an RNA extraction reagent comprising Trizol, chloroform, isopropanol, 75% ethanol, and RNase-free water.
- 5. The use according to claim 1, wherein the kit further comprises a system for reverse transcription of mRNA into cDNA and a fluorescent quantitative PCR reaction system, the system for reverse transcription of mRNA into cDNA comprises a reverse transcription complex reagent 5X RT mastermix, the reverse transcription complex reagent 5X RT mastermix comprises PrimeScriptRTase, RNase Inhibitor, Random 6 mers, Oligo dT Primer, dNTPmix, reaction Buffer;the fluorescent quantitative PCR reaction system comprises a fluorescent quantitative PCR reaction enzyme SYBR Premix Ex TaqTM.
- 6. The use according to any one of claims 1 to 5, wherein the detection step of the kit for detecting the level of PSMC2 gene comprises:a) treating and grinding the obtained fresh tissue of the glioblastoma multiforme by liquid nitrogen, and extracting RNA;b) reverse transcribing the extracted RNA into corresponding cDNA;c) performing fluorescent quantitative PCR amplification on the gene PSMC2 and the gene 18s by using the reverse transcribed cDNA;d) taking 18s as an internal reference, recording the Ct value of each reaction, and expressing the detection result as-delta Ct, wherein the delta Ct is CtPSMC2-Ct18sIn 2 of-ΔCtAs an index for evaluating the expression level of PSMC2 gene.
- 7. Wherein the Ct value is the cycle number of the fluorescence signal in each reaction tube reaching a set threshold value, CtPSMC2Is the cycle number, Ct, required for the amplification of PSMC2 expression in the reaction tube18sThe number of cycles required for expression of 18s to be amplified in the reaction tube.
- 8. The use according to any one of claims 1 to 5, wherein the kit is for grading the malignancy of glioblastoma.
- 9. The use according to any one of claims 1 to 5, wherein the kit is used for assessing the prognosis and the effect of chemotherapy in a patient with glioblastoma.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101429540A (en) * | 2007-11-09 | 2009-05-13 | 北京雅康博生物科技有限公司 | Method for real time quantitative PCR detection of mitotic cycle protein B2 gene expression |
| US20110027784A1 (en) * | 2008-01-07 | 2011-02-03 | Council Of Scientific & Industrial Research | Novel Primers for Identification of Astrocytoma, Its Grades and Glioblastoma Prognosis |
| CN109182528A (en) * | 2018-10-22 | 2019-01-11 | 中国医科大学附属第医院 | A kind of glioblastoma auxiliary diagnosis based on ITGB5 gene, prognostic evaluation kit and its application method |
-
2021
- 2021-06-28 CN CN202110719341.4A patent/CN113278702A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101429540A (en) * | 2007-11-09 | 2009-05-13 | 北京雅康博生物科技有限公司 | Method for real time quantitative PCR detection of mitotic cycle protein B2 gene expression |
| US20110027784A1 (en) * | 2008-01-07 | 2011-02-03 | Council Of Scientific & Industrial Research | Novel Primers for Identification of Astrocytoma, Its Grades and Glioblastoma Prognosis |
| CN109182528A (en) * | 2018-10-22 | 2019-01-11 | 中国医科大学附属第医院 | A kind of glioblastoma auxiliary diagnosis based on ITGB5 gene, prognostic evaluation kit and its application method |
Non-Patent Citations (3)
| Title |
|---|
| CHAO-QUN LIN等: "Effect of differential hypoxia-related gene expression on glioblastoma", RETROSPECTIVE CLINICAL RESEARCH REPORT, vol. 49, no. 5, 22 May 2021 (2021-05-22) * |
| QUENTIN KLOPFENSTEIN等: "Cell lines and immune classification of glioblastoma define patient\'s prognosis", BRITISH JOURNAL OF CANCER, vol. 120, no. 8, pages 806 - 814 * |
| 周修明;徐静;陈旭娜;何桂录;马宇强;赵华福;李维平;刘文兰;陈保东;: "表皮生长因子受体在胶质母细胞瘤中的表达及其与患者预后的关系", 广东医学, no. 08, pages 60 - 64 * |
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