CN110484624B - Gastric cancer biomarker based on peripheral blood and detection method and application thereof - Google Patents

Gastric cancer biomarker based on peripheral blood and detection method and application thereof Download PDF

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CN110484624B
CN110484624B CN201910801336.0A CN201910801336A CN110484624B CN 110484624 B CN110484624 B CN 110484624B CN 201910801336 A CN201910801336 A CN 201910801336A CN 110484624 B CN110484624 B CN 110484624B
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汪俊军
王红胜
葛李晨
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Nanjing University
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Abstract

The invention discloses a gastric cancer biomarker based on peripheral blood and a detection method and application thereof. The invention also discloses a method for detecting the RNA in the peripheral bloodm6A level to diagnose gastric cancer. The specific peripheral blood RNA m provided by the invention6A is used as an intermediate result for diagnosing the gastric cancer through single or combined related mRNA or combined serum tumor marker detection combination, and is helpful for doctors to diagnose the gastric cancer by combining clinical symptoms, medical history or other examination information.

Description

Gastric cancer biomarker based on peripheral blood and detection method and application thereof
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a gastric cancer biomarker based on peripheral blood, and a detection method and application thereof.
Background
Gastric cancer, which is a malignant tumor originating from the epithelium of the gastric mucosa, is one of the most common malignant tumors of the digestive system. In 2018, more than 100 million new gastric cancer cases are expected, and about 78.3 ten thousand death cases are respectively located at the 5 th morbidity of malignant tumors and the 3 rd mortality of diseases. Over 70% of new cases of gastric cancer are from developing countries, and nearly half of cases are from east asia. Therefore, early diagnosis of gastric cancer is important. At present, gastroscopic biopsy and detection of serum tumor-related markers are the main diagnostic tools. However, gastroscope biopsy is very traumatic, and serum tumor-related markers commonly used in clinic, such as carcinoembryonic antigen (CEA), carbohydrate antigen 199(CA199), carbohydrate antigen 224(CA224) and carbohydrate antigen 724(CA724), have the defects of low sensitivity, poor specificity and the like, and are not suitable for large-scale screening work. Therefore, an examination means with convenient examination, high sensitivity and good specificity is urgently needed.
Epigenetics is an emerging discipline, defined as heritable gene expression changes due to non-DNA sequence changes, and its main research contents include DNA methylation, histone post-translational modification, chromatin remodeling, and regulation and control modes such as microRNA, siRNA, RNA methylation, etc. Among them, RNA methylation is a new research direction in recent years, and is a leading hot spot in the field of epigenetics. In eukaryotes, m6A(N6Methylaldenosine) refers to a reversible methylation modification at position N6 on RNA adenine (A). As the chemical modification with highest abundance (more than 80%) on mRNA, each mRNA contains 3-5 m on average6The A modification site exists in a conserved sequence RRACH (R ═ G, A; H ═ A, C or U). m is6A methylation is a dynamically reversible process regulated by methylases and demethylases, METTL3, METTL14, WTAP are the core components of the methylase complex, and FTO and ALKBH5 are important demethylases. At the level of this RNAThe methylation modification mode plays a very important role in maintaining the cell function, embryonic development, stem cell self-renewal and differentiation, heat shock or DNA damage response, tumorigenesis and other pathophysiological processes, and the dissimilation of the mechanism can cause the abnormality of gene expression and the disorder of downstream signal pathways thereof, thereby causing various diseases related to gene regulation disorder.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to provide a gastric cancer biomarker based on peripheral blood and a detection method and application thereof.
The technical scheme is as follows: in a first aspect, the invention provides a gastric cancer biomarker which is RNAm in peripheral blood6A. One or more of ALKBH5, FTO, serum CEA, and serum CA 199.
Preferably, the gastric cancer biomarker is peripheral blood RNAm6A; preferably, the gastric cancer biomarker is peripheral blood RNAm6A. Marker combination of ALKBH5 and FTO, or peripheral blood RNAm6A. Marker combinations of serum CEA and serum CA 199.
The ALKBH5 and the FTO are the expression levels of ALKBH5 and the FTO in peripheral blood, in particular to the expression levels of mRNA of ALKBH5 and the FTO.
The serum CEA and serum CA199 are the level of CEA in serum and the level of CA199 in serum.
The invention provides application of gastric cancer biomarkers, namely RNA m in peripheral blood, in preparing a reagent or a kit for early diagnosis of gastric cancer6A. One or more of ALKBH5, FTO, serum CEA, and serum CA 199.
The application refers to the detection of the RNAm in the peripheral blood of a gastric cancer patient6A level, the expression level of ALKBH5, the expression level of FTO, the serum CEA level, the serum CA199 level, and a corresponding index level of a healthy population or a population of patients with benign gastric disease, wherein the corresponding index level refers to an external index level detected by the healthy population or the population of patients with benign gastric diseaseRNAm in samples of peripheral blood6A level, expression level of akbh 5, expression level of FTO, serum CEA level, serum CA199 level.
The third aspect of the invention provides the application of a reagent or a kit for detecting gastric cancer biomarkers, namely the RNAm in peripheral blood, in the preparation of a reagent or a kit for early diagnosis of gastric cancer or in the preparation of a reagent or a kit for diagnosis of gastric cancer6A. The application is to detect the RNAm in the peripheral blood of the gastric cancer patient6Comparing the A level to a control reference value, wherein the control reference value is the RNAm in a peripheral blood sample of a healthy population or a population of patients with benign gastric disease6A level.
The gastric cancer biomarkers further include one or more of ALKBH5, FTO, serum CEA, and serum CA 199.
In a fourth aspect, the present invention provides a reagent or a kit for early diagnosis of gastric cancer or diagnosis of gastric cancer, comprising detecting peripheral blood RNAm6A reagent or kit.
The fifth aspect of the present invention provides a reagent or a kit for early diagnosis of gastric cancer or diagnosis of gastric cancer, comprising:
detection of peripheral blood RNAm6A detection reagent or kit of a;
and/or a reagent or a kit for detecting the mRNA expression level of peripheral blood ALKBH 5;
and/or a reagent or a kit for detecting the mRNA expression level of peripheral blood FTO;
and/or a reagent or kit for detecting serum CEA levels;
and/or a reagent or kit for detecting serum CA199 levels.
Preferably, the reagent or kit for detecting gastric cancer further comprises a reagent or kit for extracting peripheral blood RNA.
Preferably, the reagent or kit for extracting peripheral blood RNA comprises a blood RNA stabilizer.
Preferably, the reagent or kit for early diagnosis of gastric cancer or diagnosis of gastric cancer comprises:
primer 1 shown as SEQ ID NO.1 and primer 2 shown as SEQ ID NO.2 for detecting the mRNA expression level of peripheral blood ALKBH 5;
and/or the primer 3 shown as SEQ ID NO.3 and the primer 4 shown as SEQ ID NO.4 for detecting the mRNA expression level of peripheral blood FTO.
Preferably, a reagent or kit for early diagnosis of gastric cancer or diagnosis of gastric cancer, comprising: detection of peripheral blood RNAm6A, a reagent or a kit for detecting the mRNA expression level of peripheral blood ALKBH5 and a reagent or a kit for detecting the mRNA expression level of peripheral blood FTO;
preferably, a reagent or kit for early diagnosis of gastric cancer or diagnosis of gastric cancer, comprising: detection of peripheral blood RNAm6A, a reagent or a kit for detecting CEA level in serum, and a reagent or a kit for detecting CA199 level in serum.
The sixth aspect of the invention provides a primer for detecting ALKBH5 and application of the primer for detecting FTO in preparing a reagent or a kit for early diagnosis of gastric cancer or in preparing a reagent or a kit for diagnosis of gastric cancer. Primer 1 for detecting mRNA of peripheral blood ALKBH5 is shown as SEQ ID NO.1, and primer 2 is shown as SEQ ID NO. 2; the primer 3 for detecting mRNA of peripheral blood FTO is shown as SEQ ID NO.3, and the primer 4 is shown as SEQ ID NO. 4.
The seventh aspect of the present invention provides a method for detecting and analyzing by using the reagent or kit for early diagnosis of gastric cancer:
(1) collecting and processing a peripheral blood sample from a subject;
(2) m for detecting RNA in peripheral blood sample by using kit6A methylation level;
(3) m of RNA in peripheral blood of gastric cancer patient to be detected6A methylation level, alone or in combination with any one or more of the following: mRNA expression level of ALKBH5, mRNA expression level of FTO, serum CEA level, serum CA199 level, and level or expression level of corresponding biomarker in healthy population or benign gastric disease patient population, and performing statistical analysis on peripheral blood RNAm6A、ALKBH5、FTO、CEA、CA199 as an indicator of gastric cancer detection, with parameters including threshold level, area under the subject's working characteristic curve, sensitivity, specificity, p-value, and 95% confidence interval.
Preferably, m of RNA in peripheral blood of gastric cancer patient to be detected6Comparing the level of methylation of A, the level of mRNA expression of ALKBH5 and the level of mRNA expression of FTO to the level or expression level of the corresponding biomarker in healthy subjects or subjects with benign gastric disease, and statistically analyzing the peripheral blood RNA m6A. Evaluating the performance of one or more of ALKBH5 and FTO as a gastric cancer detection index by combining, wherein the evaluated parameters comprise a critical value (cutoff value) level, an area under a working characteristic curve (AUC) of a subject, sensitivity, specificity, a p value and a 95% confidence interval;
preferably, m of RNA in peripheral blood of gastric cancer patient to be detected6Comparing the combination of one or more of A methylation level, serum CEA level and serum CA199 level with the level of the corresponding biomarker in healthy population or population of patients with benign gastric disease, and performing statistical analysis on peripheral blood RNAm6A. The combination of one or more of CEA and CA199 was evaluated for performance as an indicator of gastric cancer detection, with parameters including cutoff value (cutoff value) level, area under the subject's working characteristic curve (AUC), sensitivity, specificity, p-value, and 95% confidence interval.
Peripheral blood RNAm detected by the invention6The combination of a levels with one or more of the expression levels of akbh 5, FTO, or with one or more of serum CEA and CA199 levels, may further improve the sensitivity and specificity of the assay.
The eighth aspect of the invention provides an in vitro or in vitro use of the gastric cancer biomarker of peripheral blood as a drug candidate or a vaccine candidate, wherein the gastric cancer biomarker of peripheral blood is RNAm6A. One or more of ALKBH5, FTO CEA, and CA 199.
Preferably, the peripheral blood gastric cancer biomarker is RNAm6A is either RNAm6A in combination with one or more of ALKBH5 and FTO;
preferably, the peripheral blood gastric cancer biomarker is RNAm6A is either RNAm6A in combination with one or more of serum CEA and serum CA 199.
The drug candidate or vaccine candidate is a potential compound or biological product effective in preventing and/or treating gastric cancer.
The present invention utilizes detection of RNA m6A and detection of RNAm6The reagent or the kit suitable for clinical diagnosis and treatment is prepared by combining A with any one or more of ALKBH5, FTO, CEA and CA199, and provides a specific intermediate result and a quick and simple noninvasive detection means for realizing early detection of gastric cancer or detection of gastric cancer.
As used herein, the term "subject" refers to an individual (preferably a human) having or suspected of having a disease, or, in predicting a susceptibility, may also include healthy individuals. The term is often used interchangeably with "patient", "test subject", "treatment subject", and the like.
The term "healthy examiner" as used herein refers to a healthy population without underlying disease.
The term "benign gastric disease patient" as used herein refers to a population of other benign gastric diseases who do not have malignant gastric cancer.
Further, the data of patients with benign gastric diseases collected in the present invention refers to those patients with chronic gastritis or gastric polyps.
The invention uses reagent or reagent kit to detect peripheral blood RNA m6The A level, the mRNA expression level of peripheral blood ALHBH5 and FTO, and the serum CEA and CA199 level can be used for evaluating the gastric cancer risk of a detected object by analyzing the combined result of one or more indexes, and have important guiding significance for screening gastric cancer and subsequent diagnosis and treatment schemes.
In fact, the present invention detects peripheral blood RNA m by means of a reagent or a kit6A level, peripheral blood ALHBH5 and FTO mRNA expression level, and can be used for monitoring progression and metastasis of gastric cancerThe condition is shifted, which is helpful for disease course monitoring, operation treatment effect evaluation and prognosis judgment.
Has the advantages that: (1) the detection indexes related in the invention are as follows: peripheral blood RNA m6The level A has higher detection sensitivity and specificity in the diagnosis of gastric cancer and can be used as an excellent index for detecting gastric cancer; (2) detection indexes such as peripheral blood RNA m involved in the present invention6Level A, m6mRNA expression levels of A demethylase ALKBH5 and FTO, serum CEA and CA199 levels are different between gastric cancer patients and healthy people, and peripheral blood RNA m6The level A can be used as an excellent index for detecting the gastric cancer, and the detection can be combined with one or more of ALKBH5 and FTO or combined with one or more of CEA and CA199 to further improve the detection sensitivity and specificity, the performance of the kit is obviously superior to that of serum tumor markers CEA and CA199 used clinically at present, and the detection accuracy is greatly improved; (3) peripheral blood RNA m detected by the invention6The A level, the mRNA expression levels of ALHBH5 and FTO have correlation with the malignancy degree of the gastric cancer such as staging and metastasis, and are expected to be used as evaluation indexes for monitoring the disease course of the gastric cancer and judging prognosis; (4) the peripheral blood index detected by the invention is easier to obtain compared with other tissues, and compared with gastroscope biopsy, the invention belongs to non-invasive examination, thereby greatly facilitating the use of medical personnel and further relieving the pain of patients; (5) the detected RNA of the peripheral blood reflects the overall pathological and physiological conditions of the organism, and the detection result is accurate and detailed; (6) peripheral blood RNA extraction technology and RNA m contained in kit of the invention6The detection technology A and the fluorescence quantitative PCR technology are easy to operate and good in repeatability, and the pertinence and the practicability are improved, so that the sensitivity and the specificity of the gastric cancer detection are improved.
Drawings
FIG. 1 is a flow chart showing the operation of the gastric cancer detection kit of the present invention;
FIG. 2 shows peripheral blood RNA m6A, detecting a result; wherein, A is m of peripheral blood RNA of healthy control group (n-75), benign gastric disease group (n-30) and gastric cancer group (n-100)6A level; b is peripheral blood RNA of gastric cancer patients in stage I, stage II, stage III and stage IVm6A level; c is m of peripheral blood RNA of patients with metastatic gastric cancer and non-metastatic gastric cancer6A level; d is m of peripheral blood RNA of patients with gastric cancer (n ═ 7) before and after operation (7 days)6A level;
FIG. 3 shows the results of expression detection of ALKBH5 and FTO in peripheral blood RNA; wherein A is the expression level of ALKBH5 mRNA in peripheral blood RNA of a healthy control group and a gastric cancer group; b is the expression level of FTO mRNA in peripheral blood RNA of a healthy control group and a gastric cancer group; c is the expression level of FTO mRNA in peripheral blood RNA of patients with stage I gastric cancer and stage IV gastric cancer; d is the expression level of FTO mRNA in peripheral blood RNA of patients with metastatic gastric cancer and non-metastatic gastric cancer; e is the level of expression of ALKBH5 mRNA in peripheral blood of patients with gastric cancer (n ═ 7) before and after surgery (7 days). Table 1 shows the performance parameters of each assay index, including AUC, sensitivity%, specificity%, p-value, and 95% confidence interval.
FIG. 4 shows peripheral blood RNA m6A performance evaluation in gastric cancer detection; wherein A is peripheral blood RNA m6A, ROC curve analysis in gastric cancer patients and healthy people; b is ROC curve analysis of serum CEA in gastric cancer patients and healthy people; c is ROC curve analysis of serum CA199 in gastric cancer patients and healthy people; d is ROC curve analysis of peripheral blood ALKBH5 mRNA in gastric cancer patients and healthy people; e is ROC curve analysis of peripheral blood FTO mRNA in gastric cancer patients and healthy people; f is peripheral blood RNAm6ROC curve analysis of the combination of serum CEA and CA199 in gastric cancer patients and healthy people; g is peripheral blood RNA m6And (3) ROC curve analysis of the combination of ALKBH5 mRNA and FTO mRNA in gastric cancer patients and healthy people.
Detailed Description
The technical solution provided by the present invention is further illustrated below with reference to specific examples.
Example 1: blood sample collection and preservation
Peripheral blood samples were collected from jingling hospital affiliated with the medical college of Nanjing university, including 100 patients with gastric cancer, 30 patients with benign gastric diseases, and 75 healthy examiners.
0.5mL of fresh EDTA-K2 anticoagulated blood was taken, and 1.5-2 mL of RNAlock blood RNA stabilizer (Tiangen Biochemical technology (Beijing) Co., Ltd.) was immediately added.
Immediately covering the tube cover, turning upside down and mixing evenly for 8-10 times, and standing for more than 2 hours at room temperature. The uniformly mixed sample is stored in a low-temperature refrigerator at the temperature of minus 20 ℃.
Example 2: RNA extraction from blood samples
Step 1: when purifying a blood sample stored in a blood RNA stabilizer, the sample is first allowed to stand at room temperature or heated to room temperature in a 37 ℃ water bath. Then, the mixture is centrifuged at 6600rpm for 10min, the supernatant is removed by a pipette, and total RNA in blood is extracted by an RNAprep Pure blood total RNA extraction kit (Tiangen Biochemical technology (Beijing) Co., Ltd.), specifically referring to the kit instruction.
And detecting the concentration and purity of the RNA by using the Nanodrop2000, and performing subsequent operation.
Example 3: RNA m6Quantitative detection of A methylation
Using EpiQuik from EpigenekTMm6A RNA Methylation Quantification Kit (Colorimetric) Kit, the specific operation of which refers to the Kit instruction.
Example 4: real-time fluorescent Quantitative PCR (Quantitative real-time PCR)
Step 1: reverse transcription of RNA using PrimeScriptTMRT Master Mix (Takara) reverse transcription kit, according to its instructions for operation.
Reaction system:
Figure GDA0002232078600000071
reaction conditions are as follows: the system is mixed evenly, the liquid drop on the wall of the EP pipe is centrifuged to the bottom of the pipe by instantaneous centrifugation, the reaction is stopped by water bath at 37 ℃ for 30min and then at 85 ℃ for 5 s.
Step 2: PCR amplification with TB GREEN Premix Ex TaqTM(Takara) kit, according to its instructions for operation.
Reaction system:
Figure GDA0002232078600000072
Figure GDA0002232078600000081
PCR amplification conditions:
Stage 1：95℃，30s，1Cycle
Stage 2：95℃，10s→60℃，30s，40Cycles
Stage 3：95℃，5s→60℃，60s，1Cycle
Stage 4：50℃，30s，1Cycle
after the PCR amplification is completed, the amplification curve, the melting curve and the cycle number are calculated and analyzed on the equipment, and the melting curve is to show a single peak. If the requirements are not met, the data is not trusted. The relative expression of the gene is calculated by a delta CT method, wherein CT refers to a cycle threshold value. Wherein, the gene relative expression quantity is 2-ΔΔCT。
The real-time fluorescent quantitative PCR primer is synthesized by the Kinsley Biotechnology company Limited and has the following sequence:
Figure GDA0002232078600000082
peripheral blood RNAm of 100 patients with gastric cancer, 30 patients with benign gastric diseases, and 75 patients with health examination6The results of the test are shown in fig. 2, in which a in fig. 2 is m of peripheral blood RNA of healthy control group (n: 75), benign gastric disease group (n: 30), and gastric cancer group (n: 100)6A level; b is m of peripheral blood RNA of patients with gastric cancer at stages I, II, III and IV6A level; c is m of peripheral blood RNA of patients with metastatic gastric cancer and non-metastatic gastric cancer6A level; d is m of peripheral blood RNA of patients with gastric cancer (n ═ 7) before and after operation (7 days)6A level;
as can be seen from the results in FIG. 2, the peripheral blood RNAm in the healthy group and the benign gastric disease group in FIG. 2A6The level of A has no significant difference, and the peripheral blood RNA m of the gastric cancer group6A level is significantly highIn healthy and benign gastric disease groups; m of peripheral blood RNA is shown in FIG. 2B6Level A is related to stage of stomach cancer TNM, and m of peripheral blood RNA of patient with stage IV stomach cancer6The A level is obviously higher than that of the patients with the gastric cancer at the stage I and the stage II, which shows that the m of the RNA in the peripheral blood6The a level has a correlation with the degree of progression of gastric cancer; m of peripheral blood RNA of patients with metastatic gastric cancer is shown in FIG. 2C6M of A level compared with peripheral blood RNA of non-metastatic gastric cancer patient6The A level is obviously improved, and the peripheral blood m is prompted6The A level has a certain correlation with the metastasis of gastric cancer; in FIG. 2D, the postoperative peripheral blood m of the gastric cancer patient is shown6A levels were significantly reduced compared to preoperative, indicating peripheral blood m6The A level can be used as an index for potential postoperative prognosis judgment of gastric cancer.
The results of ALKBH5 and FTO expression detection in peripheral blood RNA of healthy examiners and gastric cancer patients are shown in FIG. 3, wherein A in FIG. 3 is the expression level of ALKBH5 mRNA in peripheral blood RNA of healthy control group and gastric cancer group; b is the expression level of FTO mRNA in peripheral blood RNA of a healthy control group and a gastric cancer group; c is the expression level of FTO mRNA in peripheral blood RNA of patients with stage I gastric cancer and stage IV gastric cancer; d is the expression level of FTO mRNA in peripheral blood RNA of patients with metastatic gastric cancer and non-metastatic gastric cancer; e is the level of expression of ALKBH5 mRNA in peripheral blood of patients with gastric cancer (n ═ 7) before and after surgery (7 days).
As can be seen from the results of fig. 3, in fig. 3A, the expression level of ALKBH5 mRNA in peripheral blood was significantly different between the healthy group and the gastric cancer group, and the expression level of ALKBH5 mRNA in the gastric cancer group was lower than that in the healthy group; in fig. 3B, the expression level of FTO mRNA in peripheral blood was significantly different between the healthy group and the gastric cancer group, in which the expression level of FTO mRNA was lower than that of the healthy group; it is shown in fig. 3C that the expression level of FTO mRNA is correlated with the degree of gastric cancer progression, with the expression level of FTO mRNA significantly lower in stage IV gastric cancer patients than in stage I gastric cancer patients; FIG. 3D shows that FTO mRNA expression levels are associated with gastric cancer metastasis, and that FTO mRNA expression levels in peripheral blood of metastatic gastric cancer patients with higher malignancy are significantly lower than those of non-metastatic gastric cancer patients; fig. 3E shows that the expression level of ALKBH5 mRNA in postoperative peripheral blood of a gastric cancer patient is significantly increased, indicating that the expression level of ALKBH5 mRNA in peripheral blood can be used as a potential indicator for postoperative prognosis determination of gastric cancer.
For the serum tumor markers CEA and CA199 detected by the subject, the peripheral blood RNA m is compared with the indexes detected by the invention6The evaluation of A in gastric cancer assay is shown in FIG. 4 and Table 1, where in FIG. 4, A is peripheral blood RNA m6A, ROC curve analysis in gastric cancer patients and healthy people; b is ROC curve analysis of serum CEA in gastric cancer patients and healthy people; c is ROC curve analysis of serum CA199 in gastric cancer patients and healthy people; d is ROC curve analysis of peripheral blood ALKBH5 mRNA in gastric cancer patients and healthy people; e is ROC curve analysis of peripheral blood FTO mRNA in gastric cancer patients and healthy people; f is peripheral blood RNA m6ROC curve analysis of the combination of serum CEA and CA199 in gastric cancer patients and healthy people; g is peripheral blood RNA m6And (3) ROC curve analysis of the combination of ALKBH5 mRNA and FTO mRNA in gastric cancer patients and healthy people. Table 1 shows the performance parameters of each assay index, including AUC, sensitivity%, specificity%, p-value, and 95% confidence interval.
As can be seen from the results of FIG. 4 and Table 1, in the healthy population and the gastric cancer patients, the peripheral blood RNA m6AUC of A is 0.929, sensitivity is 87%, and specificity is 92%; the AUC of the serum tumor markers CA199 and CEA commonly used in clinic are 0.603 and 0.694 respectively, which indicates that the peripheral blood RNA m6The diagnostic performance of A is significantly better than that of CA199 and CEA; when peripheral blood RNA m6When A is combined with CA199 and CEA, the AUC can reach 0.955, and the diagnostic performance is further improved; in addition, the AUC of ALKBH5 mRNA and FTO mRNA in peripheral blood are 0.628 and 0.661 respectively, when the RNA m in peripheral blood is6When the A is combined with ALKBH5 and FTO, the AUC can reach 0.946, and excellent diagnostic performance is shown.
TABLE 1 comparison of the Performance of the respective test indexes in gastric cancer test
Figure GDA0002232078600000101
As can be seen from the above results, the present invention utilizes a reagent or a kit for detecting peripheral blood RNA m6Level A, peripheral blood ALHBH5 and FTThe mRNA expression level of O and the serum CEA and CA199 levels are helpful for doctors to diagnose the gastric cancer by combining clinical symptoms, medical history or other examination information by analyzing the combined result of one or more indexes, and have important guiding significance for screening the gastric cancer and subsequent diagnosis and treatment schemes. Further, the invention detects the peripheral blood RNA m by using the reagent or the kit6The A level and the mRNA expression levels of peripheral blood ALHBH5 and FTO can also be used for monitoring the progression degree and the metastasis condition of gastric cancer, and are helpful for disease course monitoring, evaluation of surgical treatment effect and prognosis judgment. Detection of peripheral blood RNA m6A and related mRNA are simple and easy to implement and have excellent performance. The prepared RNA m for detecting peripheral blood6A and related mRNA kit is put into practical use, only the peripheral blood of a patient is needed, no other tissues are needed, and the peripheral blood RNA m is detected by a simplified technology6The level of A and related mRNA can expand the detection index of gastric cancer, improve the sensitivity and specificity of detection, enrich the means for detecting gastric cancer, peripheral blood RNAm6The A and related mRNA are expected to become important biomarkers for diagnosing gastric cancer, and have extremely important clinical application potential and value.
Sequence listing
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Claims (3)

1. Application of reagent for detecting gastric cancer biomarker in preparation of kit for diagnosing gastric cancer, wherein the gastric cancer biomarker is RNA m in peripheral blood6A。
2. Application of reagent for detecting gastric cancer biomarker in preparation of kit for diagnosing gastric cancer, wherein the gastric cancer biomarker is peripheral blood RNA m6A. Marker combinations of peripheral blood ALKBH5 and peripheral blood FTO.
3. Application of reagent for detecting gastric cancer biomarker in preparation of kit for diagnosing gastric cancer, wherein the gastric cancer biomarker is peripheral blood RNA m6A. Marker combinations of serum CEA and serum CA 199.
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