CN111690746A - Platelet RNA marker related to lung cancer and application thereof - Google Patents
Platelet RNA marker related to lung cancer and application thereof Download PDFInfo
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- CN111690746A CN111690746A CN202010608542.2A CN202010608542A CN111690746A CN 111690746 A CN111690746 A CN 111690746A CN 202010608542 A CN202010608542 A CN 202010608542A CN 111690746 A CN111690746 A CN 111690746A
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Abstract
The invention discloses a platelet RNA marker related to lung cancer and application thereof. A platelet RNA marker related to lung cancer is selected from any one or more of ITGA2B, FLNA, CTSW and CD 274. The application of the reagent for detecting the platelet RNA marker or the combination thereof in preparing the reagent for identifying the lung cancer diagnosis. The invention provides 4 platelet RNA markers, any one or more combinations of the 4 platelet RNA markers can effectively diagnose lung cancer, and the 4 platelet RNA markers can be applied to the preparation of lung cancer diagnostic reagents.
Description
Technical Field
The invention belongs to the field of medical diagnosis, and relates to a platelet RNA marker related to lung cancer and application thereof.
Background
Lung cancer is one of the most common malignant tumors worldwide, which has a high incidence and mortality rate, seriously affects human health, has hidden disease, lacks specificity in early clinical manifestation, and has poor prognosis due to the fact that about 70 percent of patients are in local late stage or have distant metastasis at the time of diagnosis. Currently, the gold standard for lung cancer diagnosis is still based on histopathological analysis of tumors. Tumor tissues, including fresh tissues and paraffin-embedded tissues, are the common specimens for lung cancer detection at present, but have the following problems: (1) the material is difficult to obtain. In patients who are already diagnosed with locally advanced or distant metastases, it is clinically difficult to obtain appropriate tumor tissue specimens for detection. (2) Local sampling does not reflect tumor heterogeneity. Tumor heterogeneity refers primarily to differences in tumor cells within the same tumor due to differences in tumor cell lines. (3) The materials are difficult to draw repeatedly. One-time material taking can not reflect the possible change of the tumor progress or the molecular level after treatment, and multiple material taking is needed for detection, but the pathological detection has larger wound and is difficult to realize repeated material taking. The 'liquid biopsy' based on blood provides a molecular detection method for clinic, has the characteristics of minimal invasion, real time, convenient operation, repeated material taking and the like, and overcomes the limitation of tumor tissues in material taking.
Platelets are multifunctional, anucleated cell fragments derived from bone marrow megakaryocytes, abundant next to red blood cells in peripheral blood. Platelets are involved in tumor development and metastasis, and tumor cells and their microenvironment can alter the levels of platelet RNA and proteins, either immediately or indirectly. These tumor-educated platelet functions are altered to promote tumor cell survival and metastasis in a variety of ways. Based on this, the RNA in platelets can act as a biomarker for tumor diagnosis.
Disclosure of Invention
The invention aims to provide a platelet RNA marker related to lung cancer.
It is another object of the present invention to provide the use of said markers or a combination thereof.
It is a further object of the present invention to provide reagents for detecting the markers or combinations thereof and uses thereof.
The purpose of the invention can be realized by the following technical scheme:
a platelet RNA marker related to lung cancer, which is selected from any one or more of ITGA2B, FLNA, CTSW and CD 274.
The platelet RNA marker related to the lung cancer preferably consists of CD274 and any one or more platelet RNAs selected from ITGA2B, FLNA and CTSW.
The platelet RNA marker related to lung cancer further preferably consists of ITGA2B, FLNA, CTSW and CD 274.
The invention relates to an application of a platelet RNA marker related to lung cancer as a detection target in preparing a reagent for identifying lung cancer diagnosis.
The application of the reagent for detecting the platelet RNA marker or the combination thereof in preparing the reagent for identifying the lung cancer diagnosis.
The reagent is preferably a primer, a probe or a chip for detecting the platelet RNA marker related to the lung cancer or the combination thereof.
A detection reagent for detecting the platelet RNA marker or the combination thereof.
The detection reagent for detecting the platelet RNA marker or the combination thereof is a specific primer, a probe or a chip for detecting the platelet RNA marker or the combination thereof related to the lung cancer, preferably a specific primer for detecting the platelet RNA marker or the combination thereof related to the lung cancer.
The specific primer sequences for detecting the platelet RNA markers related to the lung cancer or the combination thereof are shown as follows:
a diagnostic reagent for identifying lung cancer, which comprises a detection reagent of the platelet RNA marker related to lung cancer or the combination thereof.
Has the advantages that:
the invention provides 4 platelet RNA markers, any one or more combinations of the 4 platelet RNA markers can effectively diagnose lung cancer, and the 4 platelet RNA markers can be applied to the preparation of lung cancer diagnostic reagents.
Drawings
FIG. 1 levels of ITGA2B, FLNA, CTSW, and CD274 in platelets from lung (C) and non-lung (N) cancer patients
FIG. 2 ROC Curve analysis of the effects of ITGA2B, FLNA, CTSW, CD274 on the differentiation of lung and non-lung cancer patients
FIG. 3 ROC Curve analysis of the effects of ITGA2B, FLNA, CTSW, and CD274 in pairwise combinations on the differentiation of patients with lung and non-lung cancer
FIG. 4 ROC Curve analysis of the effects of ITGA2B, FLNA, CTSW, CD274 triple combination on differentiating lung and non-lung cancer patients
FIG. 5 ROC Curve analysis of the Effect of ITGA2B, FLNA, CTSW, CD274 in combination on the differentiation of patients with Lung cancer from non-Lung cancer
Detailed Description
Example 1 high throughput sequencing Observation of differences in RNA between 150 lung cancers and 70 normal human platelets
A first step; the separation method of the blood platelets comprises the following steps:
1. draw 10ml EDTA anticoagulation
2.1000rpm for 20min, and collecting the upper plasma
3. For the upper plasma, centrifugation at 1000rpm for 20min further removed cells that may be contained, and the pellet discarded
4. Adding immunomagnetic beads coated with anti-CD61 antibody into the platelet-rich plasma obtained in the third step to separate platelets
5. The obtained platelet RNA was extracted using Trizol and subjected to high throughput sequencing.
6. The high throughput sequencing results were analyzed using bioinformatics. From the analysis results, it can be seen that the intraplatelet RNA is significantly different between lung cancer and normal human, and the majority of RNA rises in the lung cancer-derived platelets.
7. We looked for RNAs that rose more than two-fold in lung cancer platelets and had p-value less than 0.05 and reads numbers greater than 100 as potential markers. We further prefer 20 platelet RNAs as subjects for further study and validation according to the selection criteria described above. The contents determined by high-throughput sequencing are shown in table 1.
TABLE 1 RNA with more than two-fold increase in platelet RNA high throughput sequencing and p-value less than 0.05, and reads number greater than 100
Example 2 use of platelet RNA fragments for the diagnosis of Lung cancer
To further verify the application of the 20 platelet RNA fragments in the diagnosis of lung cancer, which are preferred in the analysis of high throughput sequencing results, we targeted the 20 platelet RNA fragments, preferably one of them, and designed 20 pairs of primers for qRT-PCR, and we collected 50 cases of lung cancer and 50 normal persons. After platelet RNA was isolated using the same method as in example 1, qRT-PCR was performed, respectively, and the results are shown in Table 2 that ITGA2B, FLNA, CTSW, CD274 four platelet RNAs rose more than two-fold in lung cancer platelets and p-value was less than 0.05. The primers for these four platelet RNAs are shown in table 3.
TABLE 2 RNA with more than two-fold increase in platelet RNA qRT-PCR and p-value less than 0.05
Table 3 primers for ITGA2B, FLNA, CTSW, CD274
Subsequently, we collected 851 samples (including 736 lung cancer samples and 115 non-lung cancer specimens (including tuberculosis-induced lung nodule patients, hamartoma-induced non-lung cancer patients, etc.)) in two-three hospitals. After platelet RNA was isolated using the same method as in example 1, qRT-PCR was performed, respectively, and the results are shown in fig. 1, in which 4 RNAs were significantly different in the content of platelets in lung cancer (C) and non-lung cancer (N) patients. We then evaluated the effect of these four indicators in differentiating between lung and non-lung cancer patients using the ROC curve. The results are shown in FIG. 2, where the areas under the curves for ITGA2B, FLNA, CTSW, and CD274 are 0.7127, 0.6831, 0.7130, and 0.8209 (FIG. 2), respectively. The areas under the curves of ITGA2B, FLNA, CTSW and CD274 combined in pairs are respectively 0.717, 0.728, 0.835, 0.721, 0.831 and 0.822 (FIG. 3). The areas under the curves of the three combinations of ITGA2B, FLNA, CTSW and CD274 are respectively 0.728, 0.835, 0.834 and 0.831 (FIG. 4). When the 4 indexes were combined, the area under the curve was 0.917, which suggests that these 4 indexes can be used as biomarkers for lung cancer (fig. 5) for the diagnosis of lung cancer.
Sequence listing
<110> Zhenjiang Weisi Biotechnology Limited liability company
<120> platelet RNA marker related to lung cancer and application thereof
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Claims (10)
1. A platelet RNA marker associated with lung cancer characterized by being selected from any one or more of ITGA2B, FLNA, CTSW, CD 274.
2. The platelet RNA marker associated with lung cancer according to claim 1, wherein the platelet RNA marker associated with lung cancer consists of CD274 and any one or more platelet RNA selected from ITGA2B, FLNA, CTSW.
3. The platelet RNA marker associated with lung cancer according to claim 1, wherein the platelet RNA marker associated with lung cancer consists of ITGA2B, FLNA, CTSW and CD 274.
4. Use of the platelet RNA marker related to lung cancer according to any one of claims 1 to 3 as a detection target in preparation of a lung cancer detection reagent.
5. Use of a substance for detecting the platelet RNA marker related to lung cancer according to any one of claims 1 to 3 in the preparation of a lung cancer detection reagent.
6. The use according to claim 5, wherein the substance for detecting the platelet RNA marker associated with lung cancer according to any one of claims 1 to 3 is a specific primer, probe or chip for detecting the platelet RNA marker associated with lung cancer according to any one of claims 1 to 3.
7. A detection reagent for detecting the platelet RNA marker related to lung cancer according to any one of claims 1 to 3.
8. The detection reagent according to claim 7, wherein the detection reagent is a specific primer, probe or chip for detecting the platelet RNA marker associated with lung cancer according to any one of claims 1 to 3, preferably a primer for detecting the platelet RNA marker associated with lung cancer according to any one of claims 1 to 3.
9. The primer composition of claim 8, wherein the primers specific for detecting ITGA2B are shown in SEQ ID NO.1 and SEQ ID NO.2, the primers specific for detecting FLNA are shown in SEQ ID NO.3 and SEQ ID NO.4, the primers specific for detecting CTSW are shown in SEQ ID NO.5 and SEQ ID NO.6, and the primers specific for detecting CD274 are shown in SEQ ID NO.7 and SEQ ID NO. 8.
10. A lung cancer diagnostic kit comprising the detection reagent according to claim 6 to 8.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114438204A (en) * | 2021-12-29 | 2022-05-06 | 四川省肿瘤医院 | Lung cancer platelet molecular marker, kit and detection method |
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2020
- 2020-06-30 CN CN202010608542.2A patent/CN111690746A/en active Pending
Patent Citations (5)
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| CN103874770A (en) * | 2011-08-08 | 2014-06-18 | 卡里斯生命科学卢森堡控股有限责任公司 | Biomarker compositions and methods |
| CN104694623A (en) * | 2014-12-29 | 2015-06-10 | 常州杰傲病理诊断技术有限公司 | Plasma miRNA marker for diagnosis of lung cancer and application |
| CN105950753A (en) * | 2016-06-16 | 2016-09-21 | 朱伟 | Plasma miRNA marker relevant to lung adenocarcinoma auxiliary diagnosis and application thereof |
| CN107815495A (en) * | 2017-12-13 | 2018-03-20 | 南京医科大学 | A kind of blood plasma circular rna marker detection method related to non-small cell lung cancer |
| CN108998531A (en) * | 2018-08-31 | 2018-12-14 | 昆明医科大学第附属医院 | Lung cancer lowers long-chain non-coding RNA marker and its application |
Non-Patent Citations (4)
| Title |
|---|
| HAN XING ET AL.,: ""Development and Validation of Tumor-educated Blood Platelets Integrin Alpha 2b (ITGA2B) RNA for Diagnosis and Prognosis of Non-small-cell Lung Cancer through RNA-seq"", 《INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES》 * |
| KONRAD PAWELCZYK ET AL.,: ""Role of PD-L1 Expression in Non-Small Cell Lung Cancer and Their Prognostic Significance according to Clinicopathological Factors and Diagnostic Markers"", 《INT.J.MOL.SCI》 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114438204A (en) * | 2021-12-29 | 2022-05-06 | 四川省肿瘤医院 | Lung cancer platelet molecular marker, kit and detection method |
| CN114438204B (en) * | 2021-12-29 | 2024-03-19 | 四川省肿瘤医院 | Platelet molecular marker for lung cancer, kit and detection method |
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