Application of KAT6A in the preparation of a diagnostic kit for colorectal cancer liver metastasis

Abstract

The present invention discloses the use of KAT6A in the preparation of a diagnostic kit for colorectal cancer liver metastasis. This invention, for the first time, discovers the use of the KAT6A gene as a diagnostic marker for CRC liver metastasis in the early diagnosis or prognosis of CRC liver metastasis. Furthermore, the inventors have verified through experiments that, under the same conditions, KAT6A offers superior diagnostic advantages over CEA and AFP, currently discovered potential diagnostic markers for colorectal cancer, avoiding the occurrence of false positives. Therefore, it can be used as an early predictive indicator for colorectal cancer, for early screening and diagnosis of the disease, filling a gap in the early diagnosis of CRC liver metastasis.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology, and particularly relates to application of KAT6A in preparing a diagnostic kit for colorectal cancer liver metastasis. Background Art

[0002] Colorectal cancer (CRC) is one of the most common malignant tumors in the world. Metastasis is the main cause of death in 90% of CRC patients, among which the liver is the most common site of tumor metastasis in CRC patients. Approximately 50%-70% of CRC patients will develop liver metastasis. Once liver metastasis occurs, there is a lack of effective treatment methods, and the patient's prognosis is extremely poor, with a 5-year survival rate of only 11%, which is a huge challenge facing the clinical treatment of CRC. Therefore, the development of CRC diagnosis and the ability to effectively monitor the occurrence of CRC liver metastasis and achieve early diagnosis of CRC liver metastasis are of great significance for the treatment and screening of patients.

[0003] Although it has been reported in the prior art that carcinoembryonic antigen (CEA) can be used as a diagnostic marker for CRC and alpha-fetoprotein (AFP) can be used as a diagnostic marker for liver tumors, their specificity as early diagnostic markers for CRC liver metastasis is not strong.

[0004] Lysine acetyltransferase 6A (KAT6A), also known as MOZ and MYST3, is a histone acetyltransferase that belongs to the MYST (MOZ, Ybf2 / Sas3, Sas2, Tip60) family. KAT6A regulates cellular gene transcription, cell senescence, and T cell diversity. Its dysfunction is implicated in the development and progression of various tumors, including breast cancer, glioma, and leukemia. However, no research has publicly linked the KAT6A gene to CRC. Summary of the Invention

[0005] The prior art lacks effective and specific markers for the early diagnosis of CRC liver metastasis. In order to overcome the shortcomings of the prior art in the difficulty of early diagnosis of CRC liver metastasis, the present invention aims to provide a method for the quantitative use of KAT6A.

[0006] In order to achieve the above-mentioned object of the invention, the present invention adopts the following technical solutions:

[0007] Use of a substance for quantifying KAT6A in any of the following:

[0008] (1) Preparation of a diagnostic kit for colorectal cancer liver metastasis;

[0009] (2) Preparation of a colorectal cancer diagnostic kit;

[0010] (3) Prepare a kit to distinguish patients with colorectal cancer liver metastasis from patients without liver metastasis;

[0011] (3) Preparation of a colorectal cancer prognosis prediction kit;

[0012] (4) Preparation of a colorectal cancer tumor progression assessment kit;

[0013] (5) Preparation of a colorectal cancer overall survival assessment kit;

[0014] (6) Preparation of a colorectal cancer disease progression-free assessment kit;

[0015] (7) Prepare a colorectal cancer metastasis-free survival assessment kit.

[0016] Furthermore, the KAT6A refers to the gene numbered as Gene ID: 7994 in the NCBI database.

[0017] Furthermore, the substance for quantifying KAT6A includes at least one of products for quantifying KAT6A at the gene level and the protein level.

[0018] Furthermore, the substance for quantifying KAT6A includes at least one of a reagent and an instrument for quantifying KAT6A.

[0019] Furthermore, the reagents include: reagents for performing at least one of radioimmunoassay, indirect immunofluorescence assay, dot immunogold filtration assay, immunoblotting and enzyme-linked immunosorbent assay, polymerase chain reaction, denaturing gradient gel electrophoresis, nucleic acid typing chip detection, in situ hybridization and HRM method.

[0020] The polymerase chain reaction includes, but is not limited to, restriction fragment length polymorphism, single-strand conformation polymorphism, Taqman probe method, qPCR, competitive allele-specific PCR and allele-specific PCR.

[0021] Furthermore, the reagents include: KAT6A qPCR detection primers:

[0022] qPCRForward Primer: 5'-GGCTGGAGCTCACTGTCTC-3'

[0023] qPCRReverse Primer: 5'-TCTTATGCCGGGAGGAAGGA-3.

[0024] A system comprising a data processing device and a substance for quantifying KAT6A, wherein the system has any of the following functions:

[0025] (1) Diagnosis of colorectal cancer liver metastasis;

[0026] (2) diagnosis of colorectal cancer;

[0027] (3) distinguish patients with colorectal cancer liver metastasis from those without;

[0028] (3) Colorectal cancer prognosis prediction;

[0029] (4) Assessment of colorectal cancer tumor progression;

[0030] (5) Overall survival evaluation of colorectal cancer;

[0031] (6) colorectal cancer without disease progression assessment;

[0032] (7) Evaluation of metastasis-free survival in colorectal cancer.

[0033] Furthermore, the KAT6A refers to the gene numbered as Gene ID: 7994 in the NCBI database.

[0034] Furthermore, the data processing device is composed of a data input module, a data recording module, a data comparison module and a conclusion output module;

[0035] The data input module is configured to input the relative expression value of KAT6A in the cancerous tissue of the colorectal cancer patient to be tested;

[0036] The data recording module is configured to store the relative expression value and judgment threshold of KAT6A in the cancerous tissue of the colorectal cancer patient to be tested;

[0037] The data comparison module is configured to receive the relative expression value of KAT6A in the cancerous tissue of the colorectal cancer patient to be tested sent by the data input module, and call the judgment threshold from the data recording module to compare with the relative expression value of KAT6A in the cancerous tissue of the colorectal cancer patient to be tested;

[0038] The conclusion output module is configured to receive the comparison result sent by the data comparison module and make a judgment on the comparison result according to a predetermined judgment condition.

[0039] Furthermore, the substance for quantifying KAT6A includes at least one of products for quantifying KAT6A at the gene level and the protein level.

[0040] Furthermore, the substance for quantifying KAT6A includes at least one of a reagent and an instrument for quantifying KAT6A.

[0041] Furthermore, the reagents include: reagents for performing at least one of radioimmunoassay, indirect immunofluorescence assay, dot immunogold filtration assay, immunoblotting and enzyme-linked immunosorbent assay, polymerase chain reaction, denaturing gradient gel electrophoresis, nucleic acid typing chip detection, in situ hybridization and HRM method.

[0042] The polymerase chain reaction includes, but is not limited to, restriction fragment length polymorphism, single-strand conformation polymorphism, Taqman probe method, qPCR, competitive allele-specific PCR and allele-specific PCR.

[0043] Furthermore, the reagents include: KAT6A qPCR detection primers:

[0044] qPCRForward Primer: 5'-GGCTGGAGCTCACTGTCTC-3'

[0045] qPCRReverse Primer: 5'-TCTTATGCCGGGAGGAAGGA-3.

[0046] Using qRT-PCR and Western blotting, the inventors found that KAT6A expression increases in CRC normal adjacent tissues, primary CRC tissues, and liver metastatic CRC tissues. Analysis using the NCBI GEO database and TCGA revealed that KAT6A expression is significantly elevated in CRC tissues relative to normal adjacent CRC tissues. High KAT6A expression in CRC patients is positively correlated with CRC tumor progression. High KAT6A expression is positively correlated with overall survival, progression-free survival, and metastasis-free survival, indicating that high KAT6A expression is closely associated with poor prognosis in CRC patients. Receiver-operating characteristic (ROC) curve analysis demonstrated that KAT6A is a potential molecular marker for CRC diagnosis. Further validation was conducted in samples collected from clinical CRC patients using normal adjacent tissues, primary CRC tissues, and liver metastatic CRC tissues, further demonstrating that the KAT6A gene can serve as a diagnostic marker for CRC liver metastasis.

[0047] The present invention has the following advantages and effects compared to the prior art:

[0048] The present invention, for the first time, discovered the use of the KAT6A gene as a diagnostic marker for CRC liver metastasis in the early diagnosis or prognosis of CRC liver metastasis. Furthermore, the inventors have verified through experiments that, under the same conditions, it offers superior diagnostic advantages over CEA and AFP, currently discovered potential diagnostic markers for colorectal cancer, avoiding the occurrence of false positives. Therefore, it can be used as an early predictive indicator for colorectal cancer, for early screening and diagnosis of the disease, filling a gap in the early diagnosis of CRC liver metastasis.

[0049] 1. KAT6A overcomes the difficulty of diagnosing CRC liver metastasis in the early stage of existing technologies and provides a marker for early diagnosis of CRC liver metastasis;

[0050] 2. Under the same conditions, KAT6A has a higher diagnostic advantage than other diagnostic markers of CRC;

[0051] 3. KAT6A is a target for drug screening for colorectal cancer and can effectively screen for colorectal cancer treatment;

[0052] 4. The effectiveness and accuracy of the KAT6A gene as a predictive and diagnostic marker for colorectal cancer metastasis were 74.21%, with a 95% confidence interval of 0.6457-0.7587 and a P value of less than 0.0001. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1 The expression levels of KAT6A in CRC normal adjacent tissue (Adjacent), primary CRC tissue (Primary) and CRC liver metastases (Liver metastases); A is the RNA level; B and C are the protein levels.

[0054] Figure 2 The expression level of KAT6A in normal people (Normal) and CRC tissues (Tumor); A is the GEO database; B is the TCGA data; C and D are the CRC patient data collected clinically; E is the ROC curve.

[0055] Figure 3 Figure 2 is a study result on the correlation between KAT6A and the prognosis of CRC patients; A is the correlation between KAT6A expression and tumor progression in CRC patients; B is the correlation between KAT6A expression and overall survival in CRC patients; C is the correlation between KAT6A expression and disease-free survival in CRC patients; D is the correlation between KAT6A expression and metastasis-free survival in CRC patients; E is the clinically collected CRC patient data, and F is the ROC curve.

[0056] Figure 4 are ROC curves; A is the ROC curve of KAT6A; B is the ROC curve of CEA; C is the ROC curve of AFP.

[0057] Figure 5Figure 6 is a graph showing the validation data of KAT6A in clinical CRC samples; A is a representative graph showing the expression of KAT6A in normal adjacent tissues (Adjacent), primary CRC tissues (Primary), and liver metastases of CRC detected by immunohistochemistry; B is a statistical graph showing the expression of KAT6A in normal adjacent tissues (Adjacent), primary CRC tissues (Primary), and liver metastases of CRC; C is a graph showing the correlation between KAT6A expression and metastasis-free survival of CRC patients; D is a receiver operating characteristic (ROC) curve of KAT6A. DETAILED DESCRIPTION

[0058] The present invention will be described in further detail below with reference to examples and drawings, but the embodiments of the present invention are not limited thereto.

[0059] Unless otherwise specified, the reagents and materials used in the following examples are all commercially available products.

[0060] In the following examples, the KAT6A gene sequence used is from the NCBI database, numbered as Gene ID: 7994 (the number of its mRNA molecule: NM_006766.5).

[0061] The clinical samples used in the following examples were all from patients who visited Ganzhou Cancer Hospital between 2016 and 2021, and the experiments strictly adhered to medical ethics and the Declaration of Helsinki.

[0062] Example 1: KAT6A expression in CRC patients

[0063] 1. Experimental Methods

[0064] In this example, the clinical samples were colorectal cancer samples, including 21 normal CRC adjacent tissues, 21 primary CRC samples, and 21 liver metastasis CRC samples. The qPCR detection primers for KAT6A used were:

[0065] qPCRForward Primer: 5'-GGCTGGAGCTCACTGTCTC-3'

[0066] qPCRReverse Primer: 5'-TCTTATGCCGGGAGGAAGGA-3'.

[0067] 1. RNA Extraction and qPCR

[0068] 1) Tumor samples and normal adjacent tissues were obtained from clinical departments. Rice-sized pieces were minced with scissors and added with 1 mL of Trizol. The samples were sonicated and broken, and total RNA was extracted.

[0069] 2) Reverse transcribe the extracted RNA into cDNA using FastKing gDNA Dispelling RT SuperMix (TIAN GEN, Code No. KR118). Refer to the instruction manual for the procedure. See Table 1 for the specific reaction system and Table 2 for the reaction conditions.

[0070] Table 1. Reaction system

[0071] Reagents Usage 5xFastKing-RT SuperMix 4 μL Total RNA 1 μg <![CDATA[RNase-Free ddH2O]]> Fill to 20 μL

[0072] Table 2. Reaction conditions

[0073] temperature time 42℃ 15min 95℃ 3min

[0074] 3) qPCR detection was performed using SYBR (TIANGEN, Code No: FP207) according to the manufacturer's instructions. The amplification system is shown in Table 3, and the amplification conditions are shown in Table 4.

[0075] Table 3. Amplification system

[0076] Reagents Usage 2xSYBR 10 μL Forward Primer (10μM) 0.6μL Reverse Primer (10μM) 0.6μL cDNA 100ng <![CDATA[RNase-Free ddH2O]]> Fill to 20 μL

[0077] Table 4. Amplification conditions

[0078]

[0079] 2. Western blotting

[0080] 1) Tissue protein sample collection: Mince rice-sized tissue pieces. Add approximately 200 μL of RIPA protein lysis buffer. Ultrasonicate the lysate 12-15 times (power 3W) and centrifuge at 12,000 rpm in a pre-cooled centrifuge at 4°C for 10 min. Transfer the supernatant to a 1.5 mL EP tube.

[0081] 2) Determine the protein concentration of the samples using the BCA protein assay. Add 5 μL of 6× loading buffer and a predetermined amount of RIPA lysis solution to each sample to create a total loading volume of 20 μL. Load 30 μg of protein per sample. Denature the samples in a 100°C metal bath for 10 minutes before use.

[0082] 3) Electrophoresis: Prepare 12% separating gel for the experiment. Run the stacking gel at 100 V for 30 min. When bromophenol blue reaches the interface between the stacking gel and the separating gel, adjust the voltage to 120-140 V and run the gel for 60 min.

[0083] 4) Electrotransfer: Place the sponge, filter paper, gel, NC membrane, filter paper, and sponge on the black plate at the bottom. After all bubbles are removed, place the plate horizontally in an electrotransfer tank and perform wet transfer at 250 mA constant current for 2 hours.

[0084] 5) Blocking: According to the molecular weight of the target protein, cut the NC membrane after electroporation according to the marker display, wash it with PBST for 10 minutes, and block it in 5% skim milk for 30 minutes.

[0085] 6) Incubation with primary antibody: Place the NC membrane with the target protein in an incubation box. Quickly transfer the prepared primary antibody dilution solution to the incubation box using a pipette and completely immerse the NC membrane. Incubate at room temperature for 1 hour and then place in a 4°C refrigerator overnight.

[0086] 7) Membrane Elution and Secondary Antibody Incubation: Remove the incubation cassette, recover the primary antibody, and then wash three times with PBST (10 min each). Prepare a secondary antibody at a ratio of 1:5000, depending on the nature of the primary antibody. Incubate at room temperature in the dark for 1 hour. Wash three times with PBST (10 min each).

[0087] 8) Development and exposure: performed using ECL luminescent solution in a SYNGENE ultrasensitive chemiluminescence imager.

[0088] 2. Experimental Results

[0089] from Figure 1 As can be seen in A, the transcription level of KAT6A increases in CRC normal adjacent tissue, primary CRC tissue, and liver metastasis CRC tissue. Figure 1 As can be seen from Figures B and C, the expression of KAT6A protein levels in CRC normal adjacent cancer tissues, primary CRC tissues, and liver metastasis CRC tissues increased significantly in sequence.

[0090] Example 2: Correlation between KAT6A gene and CRC patient diagnosis

[0091] To further verify the expression of KAT6A gene in clinical CRC patients, the GEO database and TCGA database were used to analyze the expression of KAT6A gene in CRC normal adjacent tissues (n=52) and CRC cancer tissues (n=388). Figure 2 As shown in Figures A and B, KAT6A is significantly overexpressed in CRC cancer tissues relative to normal CRC adjacent tissues. The above experimental results indicate that KAT6A is highly expressed in CRC patients.

[0092] To verify the above database results, we performed immunohistochemistry (IHC) analysis on the CRC patient information collected clinically. The results are shown in Tables 5 and Figure 2As shown in Figures C and D, the results showed that KAT6A was significantly overexpressed in CRC tissues (n=56) compared with normal adjacent tissues (n=20), which was consistent with the database results. The ROC curve was used to evaluate its sensitivity and specificity in CRC diagnosis. Figure 2 As shown in Figure E, the results showed that the accuracy of KAT6A reached 84.18% (P<0.0001) within the 95% confidence interval.

[0093] Figure 1 and Figure 2 The results suggest that KAT6A can be used as a diagnostic marker for CRC and liver metastasis.

[0094] Table 5. IHC analysis results of CRC patients

[0095] sample Adjacent sample Tumor sample Tumor sample Tumor 1 2 1 12 21 8 41 3 2 2 2 12 22 8 42 3 3 2 3 12 23 8 43 2 4 2 4 12 24 8 44 2 5 2 5 12 25 8 45 6 6 2 6 12 26 6 46 6 7 2 7 12 27 6 47 6 8 2 8 12 28 6 48 6 9 2 9 12 29 6 49 8 10 3 10 9 30 6 50 8 11 3 11 9 31 6 51 0 12 3 12 9 32 6 52 0 13 3 13 9 33 4 53 1 14 3 14 9 34 4 54 1 15 4 15 9 35 4 55 2 16 4 16 9 36 4 56 2 17 4 17 9 37 4 18 4 18 9 38 4 19 0 19 9 39 4 20 0 20 9 40 3

[0096] Example 3: Correlation between KAT6A gene and prognosis of CRC patients

[0097] To analyze the relationship between KAT6A and the prognosis of CRC patients, we analyzed the correlation between KAT6A expression and tumor progression (n=122), overall survival (n=177), disease progression-free survival (n=98), and metastasis-free survival (n=125) of CRC patients in the TCGA and GEO databases.

[0098] The experimental results are as follows Figure 3 As shown in Figure 5A, KAT6A expression increases with CRC tumor progression. Figure 3 Panel B shows that higher KAT6A expression is associated with shorter overall survival in CRC patients. Figure 3 Panel C shows that higher KAT6A expression is associated with a shorter disease-free period in CRC patients. Figure 3 D in the figure shows that higher KAT6A expression is associated with shorter metastasis-free survival in CRC patients.

[0099] To verify the above database results, we analyzed the clinical data of CRC patients (n=56) and found that the higher the KAT6A expression, the shorter the overall survival of CRC patients, which is consistent with the database ( Figure 3 The sensitivity and specificity of the ROC curve in CRC prognosis were evaluated. Figure 3 The results showed that KAT6A had an accuracy of 70.11% (P<0.05) within the 95% confidence interval and could be used as a molecular marker for CRC prognosis.

[0100] Figure 3 The results suggested that high KAT6A expression was positively correlated with poor prognosis in CRC patients.

[0101] Example 4: Specificity and sensitivity of the KAT6A gene as a diagnostic marker for CRC liver metastasis

[0102] To further demonstrate the effectiveness of the KAT6A gene as a diagnostic marker for CRC liver metastasis, the inventors further analyzed the expression data for KAT6A, carcinoembryonic antigen (CEA), and alpha-fetoprotein (AFP) from the TCGA and GEO databases and constructed receiver operating characteristic (ROC) curves. The ROC curve objectively reflects the performance of a diagnostic method and includes metrics such as sensitivity, specificity, and accuracy. Using Graphpad Prism software in column mode, the KAT6A, CEA, and AFP expression data were divided into Group A (CRC without liver metastasis) and Group B (CRC with liver metastasis) for ROC curve analysis. This method was used to perform ROC curve analysis for KAT6A, CEA, and AFP simultaneously.

[0103] To study the application of KAT6A in clinical diagnosis and detection, the ROC curve was used to evaluate its sensitivity and specificity in the diagnosis of CRC liver metastasis. Figure 4 The results showed that KAT6A had an accuracy of 74.21% (P < 0.0001) within a 95% confidence interval, higher than the reported accuracy of CEA of 64.53% (P = 0.0587), making it suitable as a molecular marker for the diagnosis of CRC liver metastasis. Furthermore, compared to the liver cancer diagnostic marker AFP, which had an accuracy of only 51.15% (P = 0.8446) within a 95% confidence interval, it is not suitable for the diagnosis of CRC liver metastasis.

[0104] Example 5: Validation of the application of KAT6A gene as a diagnostic marker for CRC liver metastasis in clinical CRC patients

[0105] To verify the experimental results of KAT6A in the TCGA and GEO databases, we performed validation in normal adjacent cancer tissues, primary lesion tissues, and liver metastasis samples collected from clinical CRC patients, and collected patient follow-up information for analysis.

[0106] 1. Immunohistochemical analysis of KAT6A expression was performed in 76 CRC patients, including normal adjacent tissue (20 cases), primary lesion tissue (38 cases), and liver metastasis samples (18 cases). KAT6A expression in tumor tissue was scored based on immunohistochemical staining intensity. The scoring criteria were: the score corresponding to the percentage of positive cell area multiplied by the score corresponding to the intensity. For example, negative was assigned a score of 0, a positive cell area percentage of less than 25% was assigned a score of 1, 25%-50% was assigned a score of 2, 50%-75% was assigned a score of 3, and >75% was assigned a score of 4. Low, medium, and high staining intensities were assigned a score of 1, 2, and 3, respectively. Statistical analysis was performed by multiplying the corresponding scores and categorizing the samples into four categories: negative (score: 0), low expression (scores: 1-3), medium expression (scores: 4-8), and high expression (scores: 9-12).

[0107] 2. Based on the experimental results of KAT6A immunohistochemistry and the metastasis-free survival of CRC patients, the correlation between its expression and the metastasis-free survival of patients was analyzed, and the receiver operating characteristic (ROC) curve was used to evaluate the sensitivity and specificity of KAT6A in the diagnosis of CRC liver metastasis.

[0108] 2. Experimental Results

[0109] Immunohistochemistry showed that the expression level of KAT6A increased in CRC normal adjacent tissues, primary CRC tissues, and liver metastatic CRC tissues (Table 6, Figure 5 A and B in Figure 1). Survival analysis was performed based on KAT6A expression, and the results showed that higher KAT6A protein levels were associated with shorter metastasis-free survival ( Figure 5 The sensitivity and specificity of the CT-PCR assay in the diagnosis of CRC liver metastasis were evaluated using the ROC curve ( Figure 5 (D in Figure 1). The results showed that KAT6A had an accuracy rate of 72.29% at a 95% confidence interval (P < 0.0001), suggesting its potential as a molecular marker for the diagnosis of CRC liver metastasis. These results are consistent with those of KAT6A analyzed in the TCGA and GEO databases, further demonstrating the potential of KAT6A as a diagnostic marker for CRC liver metastasis.

[0110] Table 6. KAT6A expression levels in CRC normal adjacent tissues, primary CRC tissues, and liver metastatic CRC tissues

[0111] sample Adjacent sample Primary sample Primary sample Liver met. 1 2 1 12 21 4 1 12 2 2 2 12 22 4 2 12 3 2 3 12 23 4 3 12 4 2 4 9 24 4 4 12 5 2 5 9 25 4 5 12 6 2 6 9 26 4 6 12 7 2 7 9 27 4 7 9 8 2 8 9 28 3 8 9 9 2 9 8 29 3 9 9 10 3 10 8 30 3 10 9 11 3 11 8 31 2 11 9 12 3 12 8 32 2 12 9 13 3 13 8 33 0 13 6 14 3 14 6 34 0 14 6 15 4 15 6 35 1 15 6 16 4 16 6 36 1 16 6 17 4 17 6 37 2 17 8 18 4 18 6 38 2 18 8 19 0 19 6 20 0 20 6

[0112] Example 6: Colorectal Cancer Liver Metastasis Diagnosis System

[0113] A colorectal cancer liver metastasis diagnosis system comprising a data processing device and a substance for quantifying KAT6A;

[0114] The data processing device is composed of a data input module, a data recording module, a data comparison module and a conclusion output module;

[0115] The data input module is configured to input the relative expression value of KAT6A in the cancerous tissue of the colorectal cancer patient to be tested;

[0116] The data recording module is configured to store the relative expression value and judgment threshold of KAT6A in the cancerous tissue of the colorectal cancer patient to be tested;

[0117] The data comparison module is configured to receive the relative expression value of KAT6A in the cancerous tissue of the colorectal cancer patient to be tested sent by the data input module, and call the judgment threshold from the data recording module to compare with the relative expression value of KAT6A in the cancerous tissue of the colorectal cancer patient to be tested;

[0118] The conclusion output module is configured to receive the comparison result sent by the data comparison module and make a judgment on the comparison result according to a predetermined judgment condition.

[0119] The structure of the system for diagnosing colorectal cancer, differentiating patients with colorectal cancer liver metastasis and patients without metastasis, predicting the prognosis of colorectal cancer, evaluating the tumor progression of colorectal cancer, evaluating the overall survival of colorectal cancer, evaluating the disease-free period of colorectal cancer, and evaluating the metastasis-free survival of colorectal cancer is the same as above.

[0120] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the described embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principles of the present invention should be considered as equivalent replacement methods and are included in the scope of protection of the present invention.

Claims

1. Use of a substance for quantifying KAT6A in the preparation of a diagnostic kit for colorectal cancer liver metastasis, wherein the KAT6A refers to the gene numbered Gene ID: 7994 in the NCBI database.

2. Use of a substance for quantifying KAT6A in the preparation of a kit for distinguishing patients with colorectal cancer liver metastasis from patients without liver metastasis, wherein the KAT6A refers to the gene numbered Gene ID: 7994 in the NCBI database.

3. The use according to claim 1 or 2, characterized in that: The substance for quantifying KAT6A includes: at least one of products for quantifying KAT6A at the gene level and the protein level; The substance for quantifying KAT6A includes at least one of a reagent and an instrument for quantifying KAT6A.

4. The use according to claim 3, characterized in that: The reagents include: reagents for performing at least one of radioimmunoassay, indirect immunofluorescence, dot immunogold filtration, immunoblotting and enzyme-linked immunosorbent assay, polymerase chain reaction, denaturing gradient gel electrophoresis, nucleic acid typing chip detection, in situ hybridization and HRM method.

5. The use according to claim 4, characterized in that: The polymerase chain reaction includes restriction fragment length polymorphism, single-strand conformation polymorphism, Taqman probe method, qPCR, competitive allele-specific PCR and allele-specific PCR.

6. The use according to claim 5, characterized in that: The reagents include: KAT6A qPCR detection primers: qPCR Forward Primer: 5'-GGCTGGAGCTCACTGTCTC-3' qPCR Reverse Primer: 5'-TCTTATGCCGGGAGGAAGGA-3'.

7. A system, characterized in that: The system comprises a data processing device and a substance for quantifying KAT6A; the system is used for diagnosing colorectal cancer liver metastasis; The KAT6A refers to the gene numbered as Gene ID: 7994 in the NCBI database; The data processing device is composed of a data input module, a data recording module, a data comparison module and a conclusion output module; The data input module is configured to input the relative expression value of KAT6A in the cancerous tissue of the colorectal cancer patient to be tested; The data recording module is configured to store the relative expression value and judgment threshold of KAT6A in the cancerous tissue of the colorectal cancer patient to be tested; The data comparison module is configured to receive the relative expression value of KAT6A in the cancerous tissue of the colorectal cancer patient to be tested sent by the data input module, and call the judgment threshold from the data recording module to compare with the relative expression value of KAT6A in the cancerous tissue of the colorectal cancer patient to be tested; The conclusion output module is configured to receive the comparison result sent by the data comparison module and make a judgment on the comparison result according to a predetermined judgment condition.

8. The system according to claim 7, characterized in that: The substance for quantifying KAT6A includes: at least one of products for quantifying KAT6A at the gene level and the protein level; The substance for quantifying KAT6A includes at least one of a reagent and an instrument for quantifying KAT6A.

9. The system according to claim 8, characterized in that: The reagents include: reagents for performing at least one of radioimmunoassay, indirect immunofluorescence, dot immunogold filtration, immunoblotting and enzyme-linked immunosorbent assay, polymerase chain reaction, denaturing gradient gel electrophoresis, nucleic acid typing chip detection, in situ hybridization and HRM method.

10. The system according to claim 9, characterized in that The polymerase chain reaction includes restriction fragment length polymorphism, single-strand conformation polymorphism, Taqman probe method, qPCR, competitive allele-specific PCR and allele-specific PCR.

11. The system according to claim 10, characterized in that: The reagents include: KAT6A qPCR detection primers: qPCR Forward Primer: 5'-GGCTGGAGCTCACTGTCTC-3' qPCR Reverse Primer: 5'-TCTTATGCCGGGAGGAAGGA-3'.

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