Target TPI1 for combined treatment of liver cancer and application of target TPI1

Abstract

The invention relates to a target capable of being used for detecting the treatment response of a liver cancer patient, the target has extremely high accuracy for the response condition of the patient to ATRA + FOLFOX4 combined treatment, the response condition of the patient during the ATRA + FOLFOX4 combined treatment can be effectively judged, and the target has a wide application prospect.

Description

Technical Field

[0001] The present invention relates to the field of tumor treatment, and more specifically, to a therapeutic target for liver cancer combined with chemotherapy and its application. Background Art

[0003] Despite considerable efforts over the past 30 years, the effectiveness of systemic treatments for advanced HCC has been unsatisfactory to date. HCC has extremely high tumor heterogeneity and complex etiology, often activating multiple cancer-driving pathways simultaneously. So far, there is no effective targeted therapy to simultaneously block multiple activated signaling pathways. Due to the complex immune environment between and within tumors, immunotherapy is also inefficient against HCC, and although many phase 3 trials are ongoing, results to date have shown no improvement in efficacy compared to sorafenib in first-line immunotherapy. Chemotherapy for HCC is still under investigation, especially in Asia, although HCC is considered one of the most resistant tumors.

[0004] Encouragingly, the results of two phase III studies conducted by Qin et al. and Lyn et al. showed that FOLFOX4 (oxaliplatin + 5-fluorouracil / leucovorin) may bring some benefits and acceptable safety to Asian patients with advanced HCC. Currently, the FOLFOX4 regimen is approved for advanced HCC in China and has been used as an empirical treatment option by some physicians in the United States.

[0005] All-trans retinoic acid (ATRA) is a group of structural and functional analogs of vitamin A12. It is considered to be one of the strongest differentiation inducers and can induce differentiation of a variety of tumor cells, including stem cells. ATRA-based differentiation therapy has become the standard first-line treatment for acute promyelocytic leukemia (APL) with almost complete remission (CR). Its application in solid tumors is currently a hot topic of research. Our previous studies have shown that ATRA enhances the chemotherapy effect of cisplatin / oxaliplatin by inducing differentiation of liver cancer initiating cells and triggering tumor cell cycle arrest. However, due to the drug resistance of HCC, the current treatment strategy for HCC patients is still in the trial stage. In order to provide patients with more precise treatment strategies, it is urgent to develop more effective therapeutic targets for HCC patients. Summary of the invention

[0006] In order to overcome the defects of the prior art, the present invention provides a target for combined treatment of liver cancer and its application.

[0007] The first aspect of the present invention provides an application of a plasma biomarker in the preparation of a product for evaluating the response of liver cancer patients to combined treatment, wherein the plasma biomarker is TPI1.

[0008] In one embodiment, the combined treatment response refers to the treatment target response reflecting the patient's therapeutic efficacy as RE (CR+PR).

[0009] In one embodiment, the liver cancer treatment refers to ATRA+FOLFOX4 treatment.

[0010] In one embodiment, the product comprises a protein chip, a gene chip and / or a kit.

[0011] In a preferred embodiment, the kit is an ELISA detection kit.

[0012] In one embodiment, the product can be used for mass spectrometry detection to identify the plasma biomarkers and their expression levels through mass spectrometry, thereby evaluating the treatment response of liver cancer patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0014] Figure 1 The clinical data of 40 patients in the present invention;

[0015] Figure 2 shows the differentially expressed proteins (P val <0.05), A in Figure 2 screened any one of CRvsPD_Pre_A, CRvsPR_Pre_A, CRvsSD_Pre_A, PRvs PD_Pre_A, PRvsSD_Pre_A, REvsPD_Pre_A, REvsSD_Pre_A, SDvPD_Pre_A val <0.05 differential proteins, a total of 80 differential proteins before treatment (heat map); Figure 2B is the GO biological function enrichment analysis of 80 differential proteins related to different responses (PD, SD, PR, CR or RE) in group A (ATRA+FOLFOX4);

[0016] Figure 2C shows the KEGG biological pathway enrichment analysis of 80 pre-treatment differentially expressed proteins associated with different responses (PD, SD, PR, CR, or RE) in group A (ATRA+FOLFOX4).

[0017] Figure 3 shows 17 significantly different proteins (heat map) with adjP<0.05 or Coeff absolute value>1.5 among the 80 pre-treatment differential proteins related to different responses (PD, SD, PR, CR or RE) in group A (ATRA+FOLFOX4), including APCS, F8, SOD3, and their correlation with clinical factors of patients. Figure 3 A shows the heat map of 17 significantly different proteins with adjP<0.05 or Coeff absolute value>1.5 among the 80 pre-treatment differential proteins related to different responses (PD, SD, PR, CR or RE) in group A (ATRA+FOLFOX4); Figure 3 B shows the heat map of APCS in group A (ATRA+FOLFOX4). Figure 3C shows the correlation between APCS expression and clinical factors of patients; Figure 3D shows the expression of F8 in different response groups (PD, SD, PR, CR) of A(ATRA+FOLFOX4) group and B(FOLFOX4) group; Figure 3E shows the correlation between F8 expression and clinical factors of patients; Figure 3F shows the expression of SOD3 in different response groups (PD, SD, PR, CR) of A(ATRA+FOLFOX4) group and B(FOLFOX4); Figure 3G shows the correlation between SOD3 expression and clinical factors of patients.

[0018] Figure 4 The responses of the significantly different proteins in the A (ATRA+FOLFOX4) combined treatment group and the therapeutic targets APCS, F8, GSS, IGHV5-10-1, SOD3, and SSC5D in the corresponding regimen and efficacy;

[0019] Figure 5 The ROC curves of the 80 pre-treatment differential proteins (including APCS, F8, SOD3) or panel proteins with adjP<0.05 or Coeff absolute value>1.5 in the A(ATRA+FOLFOX4) group related to different responses (PD, SD, PR, CR or RE) are shown in Table 1. Figure 5 Middle A is the ROC curve of APCS as a diagnostic marker for the efficacy of ATRA+FOLFOX4 combined treatment of SD; Figure 5 Middle B is the ROC curve of F8 as a diagnostic marker for the efficacy of ATRA+FOLFOX4 combined therapy in PR; Figure 5 Middle C is the ROC curve of SOD3 as a diagnostic marker for CR efficacy in ATRA+FOLFOX4 combined therapy; Figure 5 D in the middle is the ROC curve of SOD3 / TTR / SSC5D / GP5 / IGKV1D-33 as a diagnostic marker for CR efficacy of ATRA+FOLFOX4 combination therapy; Figure 5Middle E is the ROC curve of TGFB1 / GSS / IGHV5-10-1 as a diagnostic marker for the efficacy of ATRA+FOLFOX4 combined therapy for PR; Figure 5 Middle F is the ROC curve of IGHD / VCAN / IGLV5-45 / IGLV5-39 as diagnostic markers for the efficacy of ATRA+FOLFOX4 combined treatment of SD / CR / PR; Figure 5 G in the middle is the ROC curve of IGHD / VCAN / IGLV5-45 / IGLV5-39 as diagnostic markers for the efficacy of ATRA+FOLFOX4 combined treatment of RE (PR / CR).

[0020] Figure 6 shows the specific therapeutic target proteins of the A (ATRA + FOLFOX4) combined treatment group. In Figure 6, A is the differential protein with any Pval < 0.05 in PostvsPre_CR_A, PostvsPre_PD_A, PostvsPre_PR_A, PostvsPre_RE_A, and PostvsPre_SD_A. There are 184 proteins with differences in the A (ATRA + FOLFOX4) group before and after treatment. Then, proteins with different differences in performance between the A (ATRA + FOLFOX4) group and the B (FOLFOX4) group before and after treatment were screened, totaling 49 proteins (heat map), which are the specific therapeutic target proteins of the A (ATRA + FOLFOX4) combined treatment group; Figure 6, B is the GO biological function enrichment analysis of the specific therapeutic target proteins of the A (ATRA + FOLFOX4) combined treatment group; Figure 6, C is the KEGG biological pathway enrichment analysis of the specific therapeutic target proteins of the A (ATRA + FOLFOX4) combined treatment group.

[0021] Figure 7 is an analysis diagram of the specific therapeutic target proteins with significant differences in group A. In Figure 7, A is a heat map of the significantly different proteins with adjP<0.05 or Coeff absolute value>1.2 among the specific therapeutic target proteins in the A (ATRA+FOLFOX4) combination treatment group, including TPI1(RE), APOC1(CR+PR+SD), ATRN(SD), B4GAT1(SD), F9(SD), ITIH4(SD), SPP2(SD), IGHG3(SD), TMEM198(SD), and (SD), F8 (SD), PFN1 (SD), SLC4A1 (SD), HPD (SD), among which there were 7 proteins with adjP < 0.05: TPI1 (RE), APOC1 (CR + PR + SD), ATRN (SD), B4GAT1 (SD), F9 (SD), ITIH4 (SD), SPP2 (SD); BN in Figure 7 shows the responses of 13 significantly different specific therapeutic target proteins in the A (ATRA + FOLFOX4) combination treatment group in the corresponding regimen and efficacy.

[0022] Figure 8 Plasma TPI1 concentrations were measured by ELISA in 60 HCC patients who received ATRA+FOLFOX4 combination therapy or FOLFOX4 therapy. DETAILED DESCRIPTION

[0023] The preferred embodiments of the present invention are described below in conjunction with the accompanying drawings. It should be understood that the preferred embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0024] Example 1 Biological samples and experimental reagents

[0025] 1. Biological samples: The samples used in this invention were from HCC patients enrolled in the Sixth Department of Hepatic Surgery of Oriental Hepatobiliary Surgery Hospital. The test samples were approved by the local ethics committee and informed consent was obtained from all subjects. A total of 80 plasma samples from 40 patients were randomly selected from the enrolled patients (each patient included two plasma samples before and after chemotherapy) for DIA proteomics analysis. The patient information is as follows: Figure 1 According to the treatment status of the patients, the patients were divided into the following groups:

[0026]

[0027] Efficacy evaluation: effective (CR complete remission, PR partial remission, SD disease stability) vs ineffective (PD disease progression)

[0028] 2. Experimental reagents:

[0029]

[0030]

[0031]

[0032]

[0033]

[0034] Example 2 Screening of plasma biomarkers of ATRA combined with FOLFOX4

[0035] 1. Collection and storage of proteomics samples

[0036] Peripheral blood samples were collected from some patients for proteomic analysis before the start of anticancer treatment and after at least one chemotherapy. Standard operating procedures were followed when collecting plasma samples. Briefly, peripheral blood samples (2 ml per person) were collected in EDTA anticoagulation tubes and centrifuged at 1,000 g / min for 15 min. Plasma was immediately transferred to a clean polypropylene tube and stored at -80°C after centrifugation.

[0037] 2. Protein Extraction, Digestion and Separation

[0038] Cell debris was removed from the plasma samples by centrifugation at 12,000 g for 10 min at 4°C. The supernatant was then transferred to a new centrifuge tube. The top 14 abundant proteins were removed using the PierceTM Top 14 Abundant Protein Depletion Spin Columns Kit (Thermo Fisher Scientific). Protein concentration was measured using the BCA kit as directed by the manufacturer.

[0039] The protein solution was reduced with 5 mM dithiothreitol at 56 °C for 30 min and then alkylated with 11 mM iodoacetamide for 15 min at room temperature in the dark. Afterwards, the protein sample was diluted by adding 100 mM TEAB to a urea concentration below 2 M. Finally, trypsin was added with a trypsin to protein mass ratio of 1:50 for the first overnight digestion and 1:100 for the second 4-hour digestion. The peptides were then desalted using a C18 SPE column.

[0040] The sample was separated into fractions by high pH reverse phase HPLC using an Agilent 300Extend C18 column (5 m particles, 4.6 mm inner diameter, 250 mm length). The wavelength was 214 nm and the column oven temperature was 35°C. After washing with 95% equilibration buffer A for 30 minutes and after the baseline was flat, the peptide sample was sent to HPLC. The peptides were separated using a gradient of 2% to 60% acetonitrile in 10 mM ammonium bicarbonate pH 10, with 1 minute collection per tube. Tubes 11 to 46 were collected, and 36 tubes were combined into 9 fractions and dried by vacuum centrifugation.

[0041] LC-MS / MS Analysis

[0042] The peptides were dissolved in mobile phase A of the HPLC and separated using an EASY-nLC 1200 HPLC system (ThermoFisher Scientific). Mobile phase A was an aqueous solution containing 0.1% formic acid and 2% acetonitrile, while mobile phase B was an aqueous solution containing 0.1% formic acid and 90% acetonitrile. The HPLC gradient was set as follows: 0-114 min, 4% of 33% of phase B; 114-117 min, 33% of 80% of phase B; 117-120 min, 80% of phase B, and the flow rate was kept constant at 500 nl / min. The peptides were separated using the HPLC system and then injected into the NSI ion source for ionization and then evaluated in the QExactiveTM HF-X (ThermoFisher Scientific).

[0043] The HPLC parameters remained unchanged from those used to create the library. Peptides were separated using an ultra-high performance liquid chromatography system and evaluated using a QExactiveTM HF-X mass spectrometer. A high-resolution Orbitrap was used to detect and analyze peptide precursor ions and their secondary fragments. The primary mass spectrometer scan range was set to 385-1200 m / z with a scan resolution of 120,000; the secondary scan resolution was set to 15,000. The data acquisition mode used a data-independent scanning (DIA) program, and peptide ions were fragmented in the HCD collision cell using a fragmentation energy of 27%, followed by secondary mass spectrometry analysis. The automatic gain control (AGC) of the secondary spectrum was set to 5E5, and the fixed initial mass was set to 200 m / z.

[0044] Protein Annotation in LC-MS / MS

[0045] The MaxQuant search engine (v.1.6.15.0) was used to process the generated MS / MS data. Tandem mass spectra were compared to the human SwissProt database (20422 entries) which was concatenated with a reverse bait database. Trypsin / P was selected as the cleavage enzyme and a maximum of two missed cleavages were allowed. The mass tolerance for precursor ions was set to 20 ppm in the first search and 5 ppm in the main search, while the mass tolerance for fragment ions was set to 0.02 Da. The FDR was adjusted to <1%.

[0046] 5. Proteomics data analysis

[0047] Protein expression data were obtained from two different groups (Group A: ATRA+FOLFOX4, Group B: FOLFOX4 alone). Samples at each baseline (before treatment) and time point (after treatment) were examined, forming 4 experimental batches (Group A - before treatment, Group A - after treatment, Group B - before treatment, Group B - after treatment). Samples of PD, SD, PR, and CR were measured in each batch.

[0048] All protein expression values ​​were normalized by calculating the relative abundance of each protein in each sample. Proteins with undetectable expression levels in more than 40 samples were removed. The differential expression of proteins was analyzed by fitting linear mixed models (LMM) using the lme4R package.

[0049] The proteomics data included several factors:

[0050] 1) Patient response (PD, SD, PR, CR),

[0051] 2) Time (before and after treatment), Day

[0052] 3) Grouping (experimental group A and chemotherapy control group B), Group

[0053] Considering the above three factors, each of which has different levels, a mixed linear regression model suitable for this case is used to conduct difference tests in different dimensions.

[0054] The linear model is as follows:

[0055] y~response+day+group+response:day:group+(1|patient_id)

[0056] The protein expression is the response variable (y), and the explanatory variables are treatment response (PD, SD, PR, and CR), time point (before and after treatment), and treatment group (group A and group B) as fixed effects. Patient ID is introduced as a random effect to allow us to take into account that the samples are paired (the same sample measured before and after treatment).

[0057] To test the differences between different responders to ATRA+FOLFOX4 therapy, we used the generalized linear hypothesis (glht) function in the multcompR package to test the following three contrasts: (1) differences in protein expression between PD, SD, PR, and CR before treatment in group A, considering the small number of possible CR patients and the addition of the PR+CR=RE group compared with PD and SD; (2) similar differences in group B, and (3) the interaction between the differences between groups A and B. To screen for possible target proteins of ATRA combined with FOLFOX4 therapy, we also tested the following three contrasts: (1) differences in protein expression before and after treatment in group A, considering the small number of possible CR patients and the addition of the PR+CR=RE group; (2) differences in group B before and after treatment, and (3) the interaction between the differences between groups A and B. To account for multiple comparisons, the obtained P values ​​were adjusted using the FDR method.

[0058] 6. ELISA detection of plasma TPI1 concentration:

[0059] The quantitative double antibody sandwich enzyme-linked immunosorbent assay technology is used to coat TPI1-specific antibodies on a solid phase carrier, add diluted plasma for incubation to form a solid phase antigen complex, and then add enzyme-labeled antibodies. The amount of enzyme on the solid phase carrier is positively correlated with the amount of TPI1 in the specimen. When the substrate is added, the enzyme in the sandwich complex catalyzes the substrate into a colored product, and the TPI1 is quantified based on the degree of color reaction and the standard curve.

[0060] 7. Experimental results:

[0061] In this clinical trial, patients treated with ATRA plus FOLFOX4 showed different responses from those treated with FOLFOX4 alone. We conducted an exploratory study to identify molecular markers of efficacy and potential targets of ATRA+FOLFOX4. We measured proteomic expression in blood samples of patients with PD, SD, PR, and CR before and after treatment in two groups, hoping to identify: (1) biomarkers that can indicate or predict response to ATRA+FOLFOX4 treatment before treatment; and (2) proteins that may be potential targets of ATRA+FOLFOX4 treatment.

[0062] 1. ATRA+FOLFOX4 plasma biomarkers:

[0063] In the plasma before ATRA+FOLFOX4 treatment, the differential proteins with any Pval<0.05 in CRvsPD_Pre_A, CRvsPR_Pre_A, CRvsSD_Pre_A, PRvsPD_Pre_A, PRvsSD_Pre_A, REvsPD_Pre_A, REvsSD_Pre_A, and SDvPD_Pre_A were screened. According to the heat map, a total of 80 differential proteins before treatment ( Figure 2A ): Among them, 35 proteins (ABI3BP, AFM, AGT, ALB, ALDOB, APCS, ASGR2, ATRN, ATXN7L2, C4A, CFHR2, DBH, ERAP2, F11, F12, F8, FABP1, FCGR3A, FCGR3B, GSS, HPD, ICAM3, IGHV1-69, IGHV3-38, IGHV5-10-1, IGKV3-20, IGLV1-51, MEGF8, ORM1, PFN1, PLGLA, PSAP, SERPINF2, SLC4A1, TGFB1) were also different in the comparison of the response between group A (ATRA+FOLFOX4) and group B (FOLFOX4) (Pval<0.05), that is, the indicative effect of the response was specific in group A (ATRA+FOLFOX4), and it was a specific diagnostic marker for the efficacy of the combined treatment of ATRA+FOLFOX4.

[0064] Compared with PD patients, proteins in SD, PR, and CR patients were specifically up-regulated or down-regulated. Based on the clustering of expression patterns, we divided the 80 significantly different proteins into the following modules. CR up-regulated, PR up-regulated, RE down-regulated, SD down-regulated, and SD up-regulated, which were enriched in different molecular functions and biological pathways ( Figure 2B and 2C ), among which the proteins significantly upregulated in PR patients were enriched in complement and coagulation-related pathways.

[0065] Figure 3 shows 17 significantly differentially expressed proteins with adjP<0.05 or Coeff absolute value>1.5 among the 80 pre-treatment differentially expressed proteins associated with different responses (PD, SD, PR, CR or RE) in group A (ATRA+FOLFOX4). Figure 3A, including APCS, F8, SOD3, TGFB1, GSS, IGHV5-10-1, IGHD, PFN1, SLC4A1, VCAN, IGLV5-45, IGLV5-39, KRT6C, TTR, SSC5D, GP5, IGKV1D-33), including APCS, F8, SOD3, and their correlation with clinical factors of patients. Among them, APCS, F8, GSS, IGHV5-10-1, SOD3, and SSC5D are also target proteins for ATRA+FOLFOX4 combined treatment. APCS was significantly overexpressed in SD patients in group A (ATRA+FOLFOX4) ( Figure 3B ), F8 was specifically upregulated in group A PR patients ( Figure 3D ), SOD3 was significantly increased in CR patients in group A (ATRA+FOLFOX4) ( Figure 3F The expression of APCS, SOD3 and F8 was associated with some clinical parameters such as PLT, TB and WBC ( Figure 3C , E, G), indicating that clinical characteristics can be used to promote the use of these proteins as predictors. The responses of the significantly different proteins in the ATRA+FOLFOX4 combined treatment group, including APCS, F8, GSS, IGHV5-10-1, SOD3, and SSC5D in the corresponding regimen and efficacy are shown in Figure 4 The expression of APCS, F8, and SOD3 in patients is a good indicator for predicting SD, PR, and CR (AUCs are 1, 1, and 0.938, respectively, Figure 5 AC). Fortunately, a panel consisting of proteins uniquely upregulated in CR and PR, SOD3 / TTR / SSC5D / GP5 / IGKV1D-33 and TGFB1 / GSS / IGHV5-10-1, may be effective in predicting CR and PR in patients ( Figure 5 D. Figure 5 E).

[0066]

[0067]

[0068] 2. Specific therapeutic target proteins for ATRA+FOLFOX4 treatment:

[0069] Filter any one of PostvsPre_CR_A,PostvsPre_PD_A,PostvsPre_PR_A,PostvsPre_RE_A,PostvsPre_SD_A val<0.05, there were 184 proteins with differences before and after treatment in group A (ATRA + FOLFOX4), and then 49 proteins with different expressions before and after treatment in group A (ATRA + FOLFOX4) and group B (FOLFOX4) were screened. Fig. 6A ), which are specific therapeutic target proteins in the A(ATRA+FOLFOX4) combination treatment group. Among them, 13 / 49 proteins were also associated with different responses before treatment (P val <0.05), that is, these 13 proteins (APCS, ATRN, ATXN7L2, F12, F8, HPD, IGLV1-51, ITIH4, MEGF8, PFN1, PLS1, SAA4, SLC4A1) are the targets of ATRA+FOLFOX4 and are also markers indicating prognosis before treatment. The differential proteins of CR, PR and SD patients are more likely to be involved in the complement and coagulation pathways ( Figure 6B -C). Among the 49 A(ATRA+FOLFOX4) combined treatment groups, 13 proteins were significantly different in specific therapeutic target proteins with adjP<0.05 or Coeff absolute value>1.2 ( Fig. 7A ), including TPI1(RE), APOC1(CR+PR+SD), ATRN(SD), B4GAT1(SD), F9(SD), ITIH4(SD), SPP2(SD), IGHG3(SD), TMEM198(SD), F8(SD), PFN1(SD), SLC4A1(SD), HPD(SD). Among them, there are 7 proteins with adjP<0.05: TPI1(RE), APOC1(CR+PR+SD), ATRN(SD), B4GAT1(SD), F9(SD), ITIH4(SD), SPP2(SD). These proteins have been clinically verified as targets for ATRA+FOLFOX4 treatment.

[0070] In patients with RE (CR+PR) in group A (ATRA+FOLFOX4), plasma TPI1 increased after treatment ( Figure 7B ). TPI1 is an isomerase that catalyzes the isomerization of glyceraldehyde-3-phosphate (G3P) and dihydroxyacetone phosphate (DHAP) during glycolysis and gluconeogenesis. TPI1 also leads to the production of methylglyoxal, a reactive cytotoxic byproduct that can modify and alter proteins, DNA, and lipids.

[0071] To further verify whether TPI1 can be used as a target for the efficacy of ATRA+FOLFOX4 combined treatment of CR+PR(RE), we selected 120 plasma samples before and after treatment from 60 HCC patients (including 30 cases in the ATRA+FOLFOX4 combined treatment group and 30 cases in the FOLFOX4 group) for plasma ELISA verification.

[0072]

[0073] ELISA results showed that the average plasma concentration of TPI1 before treatment was 131ug / L (CR) and 137ug / L (PR). The plasma concentration of TPI1 in the CR+PR group of patients treated with ATRA+FOLFOX4 combined therapy increased significantly after treatment, and the average plasma concentration of TPI1 after treatment was 171ug / L (CR) and 186ug / L (PR). However, the average plasma concentration of TPI1 in the FOLFOX4 group was slightly different before and after treatment ( Figure 8 ), and the plasma concentration of the protein did not change significantly in the PD group before and after treatment in both treatment groups. This clinical result suggests that TPI1 can be used as a therapeutic target for ATRA+FOLFOX4.

[0074] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. Use of a plasma biomarker in the preparation of a product for detecting the response of liver cancer patients to combined treatment, characterized in that: The plasma biomarker is TPI1.

2. The use according to claim 1, characterized in that The TPI1 is used to reflect the response of the liver cancer treatment target of the patient with a therapeutic effect of RE (CR+PR).

3. The use according to claim 1 or 2, characterized in that: The liver cancer treatment refers to ATRA+FOLFOX4 treatment.

4. The use according to claim 1, wherein the product comprises a protein chip, a gene chip and / or a kit.

5. The use according to claim 1, wherein the product can be used for mass spectrometry detection, and the plasma biomarkers and their expression levels are identified by mass spectrometry, thereby evaluating the response of liver cancer patients to therapeutic targets.

6. The use according to claim 4, wherein the kit is an ELISA detection kit.

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