Application of solute carrier protein SLC7A2 in preparation of reagent for predicting abnormal coagulation of non-small cell lung cancer and related product
By detecting the expression of SLC7A2 in tumor tissues of patients with non-small cell lung cancer, using SLC7A2 low-expression cell model and protein chip technology, the problem of insufficient ability to predict coagulation abnormalities in non-small cell lung cancer in the existing technology was solved, and the coagulation abnormalities and thrombosis were achieved. Efficient prediction of risks provides important diagnostic and therapeutic guidance.
Patent Information
- Application Number
- CN202411673883.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-21
- Publication Date
- 2025-01-14
AI Technical Summary
The prior art has insufficient predictive ability to predict coagulation abnormalities in patients with non-small cell lung cancer, especially in patients with normal coagulation function, which cannot effectively predict the risk of future coagulation abnormalities and thrombosis.
The solute carrier protein SLC7A2 was used as a predictive indicator to detect the expression of SLC7A2 in tumor tissues of patients with non-small cell lung cancer, and a stable cell line with low expression of SLC7A2 was constructed to form a cell model. Combined with protein chip technology, SLC7A2 expression was detected to predict coagulation abnormalities.
It significantly improves the predictive ability of coagulation abnormalities and thrombosis in patients with non-small cell lung cancer, prevents the occurrence of venous thromboembolic disease in advance, and provides a more effective molecular indicator for diagnosis and treatment guidance.
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Figure CN119310284A_ABST
Abstract
Description
Technical Field
[0001] The present application belongs to the technical field of biological prediction reagents, and specifically relates to the application of a solute carrier protein SLC7A2 in the preparation of a reagent for predicting coagulation abnormalities in non-small cell lung cancer and related products. Background Art
[0002] Non-small-cell lung cancer (NSCLC) accounts for more than 80% of various lung cancer types. Studies have shown that more than half of patients with non-small-cell lung cancer have coagulation abnormalities. The risk of venous thromboembolism in NSCLC patients is more than double that of small-cell lung cancer, and it is also an important cause of death in patients. Therefore, it is of great significance to find coagulation sensitivity indicators for NSCLC patients.
[0003] At present, the coagulation function of patients is mainly assessed clinically by testing the six coagulation indicators. Although the six coagulation indicators can well reflect the coagulation status of NSCLC patients, they are insufficient in predicting the possible coagulation abnormalities and thrombosis in NSCLC patients with normal coagulation. In addition, there are studies that predict the risk of hypercoagulation in lung cancer patients by combining thromboelastography and coagulation indicators, and there are also studies that predict the risk of coagulation abnormalities in lung cancer patients by detecting the concentrations of new thrombus markers such as thrombomodulin (THBD) and thrombin-antithrombin complex (TAT) in the patient's serum. However, when the above indicators are tested, the patient's coagulation function has already become abnormal, and the predictive function has certain limitations. Summary of the invention
[0004] In order to solve the technical problem that the existing methods for detecting abnormal coagulation function in lung cancer patients have poor predictive ability, the present application provides an application of a solute carrier protein SLC7A2 in the preparation of a reagent for predicting abnormal coagulation function in non-small cell lung cancer and related products.
[0005] In order to achieve the above objectives, this application adopts the following technical solutions: In a first aspect, the present application proposes the use of the solute carrier protein SLC7A2 in the preparation of a reagent / kit for predicting coagulation abnormalities in non-small cell lung cancer.
[0006] Furthermore, the reagent / kit is a predictive reagent / kit for detecting SLC7A2 expression in tumor tissue of patients with non-small cell lung cancer.
[0007] Furthermore, SLC7A2 expression in tumor tissues of patients with non-small cell lung cancer was inversely correlated with the probability of abnormal coagulation in the patients.
[0008] Furthermore, the method for detecting SLC7A2 expression in tumor tissue includes: The fibrinogen γ-chain and pro-β-chain, serine protease inhibitor E2 and serine protease inhibitor A1 were down-regulated in the patients, the role of thrombomodulin in inhibiting coagulation was up-regulated, and the expression of SLC7A2 in tumor tissues was detected.
[0009] Furthermore, the reagent is a protein chip.
[0010] Furthermore, by constructing a stable cell line with low expression of SLC7A2, a cell model was formed to predict coagulation abnormalities in non-small cell lung cancer.
[0011] In a second aspect, the present application proposes a reagent / kit for predicting coagulation abnormalities in non-small cell lung cancer, wherein the reagent / kit comprises a solute carrier protein SLC7A2.
[0012] Furthermore, the reagent / kit predicts the possibility of coagulation abnormality in patients with non-small cell lung cancer by detecting whether SLC7A2 expression in tumor tissues of patients with non-small cell lung cancer is negative.
[0013] In a third aspect, the present application proposes a protein chip for predicting coagulation abnormalities in non-small cell lung cancer, comprising a solid phase carrier and a non-small cell lung cancer coagulation abnormality protein detection point located on the solid phase carrier, wherein the solute carrier protein SLC7A2 is fixed on the protein detection point.
[0014] Furthermore, the protein chip predicted the possibility of coagulation abnormalities in patients with non-small cell lung cancer by detecting whether SLC7A2 expression in the tumor tissue of patients with non-small cell lung cancer was negative.
[0015] Compared with the prior art, this application has the following beneficial effects: This application discloses for the first time the application of the solute carrier protein SLC7A2 in predicting abnormal coagulation in non-small cell lung cancer, including the preparation of prediction reagents / kits, and protein chips for predicting abnormal coagulation in non-small cell lung cancer. By studying the relationship between the expression of the solute carrier protein SLC7A2 and abnormal coagulation in non-small cell lung cancer, the strong correlation between the expression of the solute carrier protein SLC7A2 and the probability of abnormal coagulation in non-small cell lung cancer is revealed, and its guiding role in the prediction, research, diagnosis and treatment of abnormal coagulation in non-small cell lung cancer is clarified. It is of great significance to more effectively predict the molecular indicators of abnormal coagulation and thrombosis in patients with non-small cell lung cancer. BRIEF DESCRIPTION OF THE DRAWINGS
[0016] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present application and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without paying creative work.
[0017] Figure 1 Schematic diagram of the experimental results of SLC7A2 in the examples of the present application; wherein A and B indicate the successful construction of the SLC7A2 low-expressing cell line, A is the mRNA expression of SLC7A2 detected by qPCR in A549-shRNA-Control and A549-shRNA-SLC7A2 cells, and B is Western Blot detection of SLC7A2 protein expression in A549-shRNA-Control and A549-shRNA-SLC7A2 cells, C is a schematic diagram of qPCR detection of FGG mRNA expression in A549-shRNA-Control and A549-shRNA-SLC7A2 cells, D is a schematic diagram of qPCR detection of FGB mRNA expression in A549-shRNA-Control and A549-shRNA-SLC7A2 cells, E is a schematic diagram of qPCR detection of THBD mRNA expression in A549-shRNA-Control and A549-shRNA-SLC7A2 cells, F is a schematic diagram of qPCR detection of SERPINA2 mRNA expression in A549-shRNA-Control and A549-shRNA-SLC7A2 cells, G is a schematic diagram of qPCR detection of SERPINA1 mRNA expression in A549-shRNA-Control and A549-shRNA-SLC7A2 cells, and H is Western Schematic diagram of Blot detection of SLC7A2, FGG and FGB protein expression in A549-shRNA-Control and A549-shRNA-SLC7A2 cells. DETAILED DESCRIPTION
[0018] In order to make the purpose, technical solution and advantages of the embodiments of the present application clearer, the technical solution in the embodiments of the present application will be clearly and completely described below in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments. The components of the embodiments of the present application described and shown in the drawings here can be arranged and designed in various different configurations.
[0019] Therefore, the following detailed description of the embodiments of the present application provided in the accompanying drawings is not intended to limit the scope of the present application for which protection is sought, but merely represents selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in the field without creative work are within the scope of protection of the present application.
[0020] Compared with small cell lung cancer, patients with non-small cell lung cancer have a significantly higher risk of venous thromboembolism than patients with small cell lung cancer, which may be related to the hypercoagulable state of blood, tumor-related factors and some complications during treatment in patients with non-small cell lung cancer. Venous thromboembolism not only increases the difficulty of treatment for patients with non-small cell lung cancer, but may also become an important cause of death in patients. Therefore, it is particularly important to monitor and manage the coagulation function of patients with non-small cell lung cancer. If the abnormal coagulation function of patients with non-small cell lung cancer can be predicted in the early stage, intervention measures can be taken in time to prevent the occurrence of venous thromboembolism. It can also serve as a reference for evaluating the treatment effect and prognosis of patients with non-small cell lung cancer.
[0021] The present invention studies the relationship between the expression of solute carrier protein SLC7A2 and the probability of abnormal coagulation in non-small cell lung cancer, reveals the strong correlation between the expression of solute carrier protein SLC7A2 and abnormal coagulation in non-small cell lung cancer, and clarifies the guiding role of solute carrier protein SLC7A2 expression in disease diagnosis, research and treatment.
[0022] Currently, there are two main methods to evaluate the coagulation function of patients with non-small cell lung cancer: (1) Evaluation through six coagulation indicators: The six coagulation indicators mainly include thrombin time (TT), activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen (FIB), fibrinogen degradation product (FDP) and plasma D-dimer (D-Dimer). Since the coagulation process is a complex physiological process involving the interaction of multiple coagulation factors, platelets and vascular walls, the abnormality of a single or a few coagulation indicators may not be sufficient to fully reflect the overall state of coagulation function. Therefore, although the six coagulation indicators are of great value in evaluating the coagulation function of NSCLC patients, they are indeed insufficient in predicting the possible coagulation abnormalities and thrombosis in patients with normal coagulation function in the future.
[0023] (2) Predicting the risk of hypercoagulation in lung cancer patients by combining thromboelastography and coagulation indices, or predicting the risk of abnormal coagulation in lung cancer patients by detecting the concentration of new thrombus markers such as thrombomodulin (THBD) and thrombin-antithrombin complex (TAT) in the patient's serum. Similar to the six coagulation tests, these indicators can only be detected after the patient has abnormal coagulation function and cannot be used for prediction.
[0024] The solute carrier protein SLC7A2 (Solute carrier family 7 member 2) is an important membrane protein and a member of the solute carrier family 7 (SLC7). The protein encoded by SLC7A2 is a cationic amino acid transporter and belongs to the APC (amino acid-polyamine-organic cation) transporter family. This protein is responsible for the cellular uptake of basic amino acids such as arginine, lysine and ornithine. It plays a key role in the metabolism and transport of amino acids and participates in maintaining the balance of amino acids in cells. SLC7A2 is widely expressed in various tissues, including the liver and thyroid gland. The protein is mainly located on the cell membrane and is a component of the membrane, participating in the exchange of substances between cells.
[0025] This application proposes that the solute carrier protein gene SLC7A2 can better predict coagulation abnormalities and thrombosis in NSCLC patients, and prevent the occurrence of venous thromboembolism in NSCLC patients in advance. In NSCLC tissues, patients with negative SLC7A2 expression are more likely to have coagulation abnormalities.
[0026] In addition, the present application also constructs a stable cell line with low expression of SLC7A2 to form a cell model for predicting coagulation abnormalities in non-small cell lung cancer. As an example, the expression of SLC7A2 in A549 cells can be reduced by lentiviral infection.
[0027] Among them, lentiviral infection is an efficient and stable method for constructing stable cell lines. Lentivirus vectors can effectively integrate exogenous genes into host chromosomes to achieve persistent expression. By designing shRNA sequences targeting SLC7A2 and constructing lentiviral interference vectors, the expression level of SLC7A2 can be significantly reduced.
[0028] In practical applications, specific shRNA sequences targeting the SLC7A2 gene can be designed and cloned into a lentiviral interference vector. The constructed lentiviral interference vector and the auxiliary plasmid are transfected into 293T cells together, and lentiviral particles are packaged and then filtered with a 0.22 μm filter for subsequent use. A549 cells and lentiviral solution are mixed on a shaker at room temperature for 1 hour and then inoculated into a culture plate. Fresh culture medium is replaced after 8-12 hours, and stable expression cell lines are screened according to the resistance gene carried by the lentiviral vector. After multiple rounds of screening and subculture, a stable A549 cell line with low expression of SLC7A2 can be obtained.
[0029] The constructed A549 cell line with low SLC7A2 expression can be used as a cell model to study abnormal coagulation in NSCLC. By comparing the differences in coagulation-related indicators between the SLC7A2 low-expressing cell line and the normal-expressing cell line, the mechanism of action of the SLC7A2 gene in abnormal coagulation in NSCLC can be explored.
[0030] This application also studies the coagulation-related genes regulated by SLC7A2, including FGG, FGB, SERPINE2, SERPINA1 and THBD. Through analysis, it was found that the expression of DD dimer in patients with negative SLC7A2 expression in NSCLC tumor specimens was significantly increased, and the prothrombin time PT was significantly shortened. After knocking down the expression of SLC7A2 in NSCLC cell lines, the expression of coagulation-related genes FGG, FGB, SERPINE2, SERPINA1 increased significantly, and THBD decreased significantly, proving that SLC7A2 regulates coagulation in NSCLC patients. By detecting the expression of SLC7A2 in NSCLC specimens, the possibility of coagulation abnormalities in NSCLC patients can be predicted, and early intervention can be made to reduce the occurrence of NSCLC venous thromboembolism, which has good clinical application development potential. Therefore, as a method for predicting coagulation abnormalities in NSCLC patients, the expression of SLC7A2 in tumor tissues of NSCLC patients can be detected.
[0031] As shown in Table 1 below, the clinical data comparison of positive and negative expression of SLC7A2 in tumor tissues of NSCLC patients is reflected. SLC7A2 was stained in tumor tissues of NSCLC patients using immunohistochemistry, and clinical patients were grouped according to the negative and positive expression of SLC7A2, divided into SLC7A2 positive group and SLC7A2 negative group. The clinical data of the two groups of NSCLC patients were then compared. The comparison results showed that there were a total of 4 statistically significant expression differences between SLC7A2 positive and negative patients. Two of them were present in the 6 coagulation-related indicators: the DD dimer index of patients with positive expression of SLC7A2 was significantly lower than that of negative patients, and the PT index was significantly higher than that of negative patients, proving that the expression of SLC7A2 is closely related to coagulation abnormalities in patients with non-small cell lung cancer.
[0032] Table 1 Comparison of clinical data on positive and negative expression of SLC7A2 in tumor tissues of NSCLC patients
[0033] Figure 1The figure shows that SLC7A2 upregulates the expression of coagulation-related genes FGG, FGB, SERPINE2 and SERPINA1, and downregulates the expression of THBD. In order to verify the relationship between SLC7A2 and coagulation, and further explore the mechanism of the relationship between SLC7A2 and coagulation, the expression of SLC7A2 was knocked down in A549 cells in the experiment, and then qPCR and Western Blot were used to observe the expression of the above coagulation genes and SLC7A2. The results showed that SLC7A2 can inhibit the expression of coagulation genes FGG, FGB, SERPINE2 and SERPINA1, and promote the expression of THBD. This result, on the one hand, verifies the close relationship between SLC7A2 and coagulation, and on the other hand, preliminarily explores the mechanism of their relationship.
[0034] Figure 1 A and B in the figure can show that the experiment has verified that the A549-shRNA-SLC7A2 cell line has been successfully constructed. C to G show that qPCR detects the mRNA expression of FGG, FGB, SERPINE2, SERPINA1 and THBD in A549-shRNA-Control and A549-shRNA-SLC7A2 cells. H shows that Western Blot verifies the protein expression of SLC7A2, FGG and FGB in A549-shRNA-Control and A549-shRNA-SLC7A2 cells.
[0035] The following experiments were further conducted: The expression of SLC7A2 in tumor tissues of NSCLC patients in Table 1 was compared with clinical data: the tumor tissues of NSCLC patients were dehydrated, embedded, sliced and dewaxed, and then the EDTA antigen repair solution was repaired in a microwave oven for 20 minutes. After the specimen was cooled, the calf serum was blocked at room temperature for 1 hour, and the SLC7A2 antibody (SLC7A2: primary antibody dilution = 1:200) was incubated at 4°C overnight. The next day, it was rinsed with distilled water 3 times, 10 minutes each time, and then incubated with secondary antibody at 37°C for 1 hour. Then DAB color development, hematoxylin staining, dehydration and sealing were performed for reading. Among them, the immunohistochemistry scoring standard used the German semi-quantitative scoring system to score the expression of SLC7A2.
[0036] Implementation Figure 1 SLC7A2 upregulates the expression of coagulation-related genes FGG, FGB, SERPINE2 and SERPINA1, and downregulates the expression of THBD.
[0037] Then do the following: 1. Construction of A549-shRNA-SLC7A2 stable cell line.
[0038] 1. Shanghai GeneCare Gene Co., Ltd. designed and synthesized GV654-shRNA-SLC7A2. The sequence of SLC7A2-RNAi is gcTGGGTTTGTGAAAGGAAAT.
[0039] 2. Packaging of lentivirus.
[0040] 24 h before transfection, HEK-293T cells were seeded in a 10 cm dish (cell number 1X10 5 / dish), and the next day, when the cell confluence reached 60% before transfection, DMEM medium was replaced. 20 μg of GV654-shRNA-SLC7A2 plasmid / control plasmid and 25 μl of Lenti-Easy Packaging Mix (Shanghai Jikai Gene Co., Ltd.) were added to 1.5 ml of DMEM medium and mixed. Then, 60 μl of Lipofectamine 2000 was added to 1.5 ml of DMEM medium and mixed. After standing at room temperature for 5 minutes, the Lipofectamine 2000 mixture was added dropwise to the mixed plasmid solution. After mixing, it was allowed to stand at room temperature for 20 minutes. The above mixture was added to the HEK-293T cells that had been replaced with medium. After incubation at 37°C for 6 hours, fresh DMEM medium with 10% FBS was replaced; after replacing the fresh medium, incubate for 24 hours, collect the supernatant (containing viral particles) and store at 4°C; after re-adding fresh medium and incubating for 24 hours, collect and combine the supernatant, filter the supernatant with a 0.22 μm microporous filter membrane, and store it in a -80° refrigerator.
[0041] 3. Infect target cells with lentivirus.
[0042] Add 2 ml of the above supernatant (containing virus particles) to 2 ml of fresh DMEM medium containing 10% FBS, and add 10 μl of polybrene solution at the same time. Mix well and add 10 4 A549 cells were placed on a shaker at room temperature for 1 hour and then inoculated into a 6 cm cell culture dish. After 12 hours, fresh DMEM medium with 10% FBS was replaced. When the cells grew to 80%, an appropriate concentration of G418 was added to select the cells, and the medium containing G418 was replaced every 3 days until the cells stopped dying. After that, 1 / 2 concentration of G418 was used to maintain the cultured cells until they were full for subsequent experiments. The above cells were named A549-shRNA-Control and A549-shRNA-SLC7A2.
[0043] 4. Verify the successful construction of the cell line by fluorescence quantitative PCR (qPCR).
[0044] Extraction of total mRNA from A549-shRNA-SLC7A2 and A549-shRNA-Control cells. The total mRNA of the two cells was extracted using the Animal Tissue / Cell Total mRNA Extraction Kit (Tiangen Biochemical Technology Co., Ltd.). The following methods can be used: ① Add 350 ml of lysis buffer RLA and 10 μl of proteinase K to the cells inoculated in a 6-well plate, mix well by pipetting, and place at room temperature for 5 minutes; ② Centrifuge at 12,000 rpm for 2 minutes, take the supernatant, add it to the genomic DNA removal column, centrifuge at 12,000 rpm for 30 seconds, and keep the filtrate; ③ Add an equal volume of 70% ethanol to the above filtrate, mix well and add to the RNase-Free adsorption column CR4, centrifuge at 12,000 rpm for 30 seconds, and discard the waste liquid; ④ Add 700 μl of deproteinized solution RW3 to the adsorption column CR4, centrifuge at 12,000 rpm for 30 seconds, and discard the waste liquid; ⑤ Add 500 μl of rinsing solution RW to the adsorption column CR4, let it stand at room temperature for 2 minutes, then centrifuge at 12,000 rpm for 30 seconds and discard the waste liquid; ⑥Repeat the above steps.
[0045] ⑦ Centrifuge the empty tube at 12,000 rpm for 2 minutes, discard the waste liquid, and place the adsorption column CR4 at room temperature for two minutes to dry the residual liquid; ⑧ Transfer the adsorption column CR4 into a new RNase-Free centrifuge tube, add 30 μl RNase-Free ddH2O, leave it at room temperature for 2 minutes, and then centrifuge it at 12,000 rpm for 2 minutes to obtain total RNA.
[0046] 2. Reverse transcribe the harvested RNA to obtain cDNA.
[0047] 1. Prepare the mixture according to the genomic DNA removal system in Table 2 and mix thoroughly. Centrifuge briefly and incubate at 42°C for 3 min. Then place on ice.
[0048] Table 2 gDNA removal reaction system
[0049] 2. Prepare the mixed solution according to the reverse transcription reaction system in Table 3.
[0050] Table 3 Reverse transcription reaction system
[0051] 3. Add the Mix from the reverse transcription reaction to the reaction solution from the gDNA removal step, mix thoroughly, and incubate at 42°C for 15 min.
[0052] 4. Incubate at 95°C for 3 min and place on ice. The obtained cDNA can be used for subsequent experiments or stored at low temperature.
[0053] The qPCR system was prepared according to Table 4, and the primer list is shown in Table 5. The reaction conditions were: 95℃, 30 seconds; 95℃, 5 seconds; 60℃, 34 seconds; 95℃, 15 seconds; 60℃, 1 minute; 95℃, 15 seconds.
[0054] Table 4 qPCR system list
[0055] Table 5 Primer list
[0056] qPCR results showed that the expression of SLC7A2 in A549-shRNA-SLC7A2 cells was significantly lower than that in A549-shRNA-Control cells, proving that the cell line was successfully constructed from the perspective of mRNA ( Figure 1 A), and after reducing SLC7A2 expression, FGG ( Figure 1 C) FGB Figure 1 D) SERPINA1 Figure 1 G) and SERPINE2 ( Figure 1 F) mRNA expression increased, THBD ( Figure 1 E) decreased expression.
[0057] 5. Use Western Blot to verify the successful construction of the cell line.
[0058] A549-shRNA-SLC7A2 and A549-shRNA-Control cells were inoculated in 6 cm cell culture dishes. The next day, they were rinsed three times with pre-cooled PBS, and then 200 μl of RAPI lysis buffer and 20 μl of PMSF were added and mixed. The mixture was lysed on ice for 30 minutes, and then centrifuged at 12,000 rpm for 15 minutes at 4°C. The supernatant was harvested as the total protein. 5× protein loading buffer was added to the total protein, and the Western Blot experiment was performed after incubation at 100°C for 10 minutes.
[0059] Among them, the Western Blot experiment can be specifically performed using the following methods: (1) Prepare 10% SDS-PAGE separation gel and 5% SDS-PAGE stacking gel. Add total protein and loading marker to the stacking gel and run electrophoresis at 80V for about 3 hours.
[0060] (2) Transfer the membrane at 200 mA for 2 hours.
[0061] (3) Rinse the transferred membrane briefly with TBST and then block it with 5% skimmed milk powder at room temperature for 1 hour.
[0062] (4) After blocking, incubate with primary antibody overnight (primary antibody concentration is 1:1000).
[0063] (5) The next day, wash the membrane three times with TBST for 10 minutes each time, and then incubate with secondary antibody for 2 hours at room temperature. Wash the membrane 3×10min. (6) Exposure.
[0064] Western Blot results showed that the protein expression of SLC7A2 in A549-shRNA-SLC7A2 cells was significantly lower than that in A549-shRNA-Control cells, which proved that the cell line was successfully constructed from the perspective of protein. In addition, after reducing the expression of SLC7A2, the protein expressions of FGG and FGB increased ( Figure 1 H). This result is consistent with the qPCR result, which can prove that SLC7A2 is closely related to coagulation and coagulation-related genes, providing strong evidence for SLC7A2 to predict coagulation abnormalities in non-small cell lung cancer.
[0065] This application uses the solute carrier protein SLC7A2 to downregulate fibrinogen FGG and FGB, serine protease inhibitors E2 (SERPINE2) and A1 (SERPINA1) in patients with non-small cell lung cancer, and upregulates the role of thrombomodulin (THBD) in inhibiting coagulation, and detects the expression of SLC7A2 to predict the possible coagulation abnormalities and thrombosis risks in patients with non-small cell lung cancer. In patients with non-small cell lung cancer, there are significant differences in DD dimer and prothrombin time between SLC7A2-negative and positive specimens. In in vitro cell validation experiments, the expression of coagulation-related genes FGG, FGB, SERPINE2 and SERPINA1 in cell lines with decreased SLC7A2 expression increased significantly, and THBD expression decreased significantly, proving that SLC7A2 is closely related to coagulation and has good potential for predicting thrombosis.
[0066] The above are only preferred embodiments of the present application and are not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims
1. Application of solute carrier protein SLC7A2 in the preparation of reagents / kits for predicting coagulation abnormalities in non-small cell lung cancer.
2. The use according to claim 1, characterized in that The reagent / kit is a predictive reagent / kit for detecting SLC7A2 expression in tumor tissue of patients with non-small cell lung cancer.
3. The use according to claim 2, characterized in that The expression of SLC7A2 in tumor tissues of patients with non-small cell lung cancer is inversely correlated with the probability of abnormal coagulation in these patients.
4. The use according to claim 3, characterized in that Methods for detecting SLC7A2 expression in tumor tissues include: The fibrinogen γ-chain and pro-β-chain, serine protease inhibitor E2 and serine protease inhibitor A1 were down-regulated in the patients, the role of thrombomodulin in inhibiting coagulation was up-regulated, and the expression of SLC7A2 in tumor tissues was detected.
5. The use according to any one of claims 1 to 4, characterized in that: The reagent is a protein chip.
6. The use according to any one of claims 1 to 4, characterized in that: By constructing a stable cell line with low expression of SLC7A2, a cell model was formed to predict coagulation abnormalities in non-small cell lung cancer.
7. A reagent / kit for predicting abnormal coagulation in non-small cell lung cancer, characterized in that: The reagent / kit contains the solute carrier protein SLC7A2.
8. The reagent / kit for predicting abnormal coagulation in non-small cell lung cancer according to claim 7, characterized in that: The reagent / kit predicts the possibility of coagulation abnormality in patients with non-small cell lung cancer by detecting whether SLC7A2 expression in tumor tissue of patients with non-small cell lung cancer is negative.
9. A protein chip for predicting abnormal coagulation in non-small cell lung cancer, characterized in that: The invention comprises a solid phase carrier and a non-small cell lung cancer coagulation abnormality protein detection point located on the solid phase carrier, and a solute carrier protein SLC7A2 is fixed on the protein detection point.
10. The protein chip according to claim 9, characterized in that The protein chip predicts the possibility of coagulation abnormalities in patients with non-small cell lung cancer by detecting whether SLC7A2 expression in the tumor tissue of patients with non-small cell lung cancer is negative.