Application of CUL7 as a biomarker in the prognostic evaluation of colon adenocarcinoma

Abstract

The present invention belongs to the field of biotechnology and discloses the use of CUL7 as a biomarker for the prognosis assessment of colorectal adenocarcinoma, specifically disclosing the use of CUL7 as a biomarker. The present invention uses bioinformatics and tissue microarrays (TMA) to explore the relationship between CUL7 expression and the prognosis and clinicopathological characteristics of colorectal adenocarcinoma (COAD). The results show that CUL7 expression is significantly increased in COAD tumor tissue compared to normal tissue. The K-M curve shows that in COAD, high CUL7 expression is associated with decreased survival, suggesting that CUL7 may be used as a biomarker for early screening or prognosis of colorectal adenocarcinoma and as a therapeutic target in clinical trials.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology, and particularly relates to the application of CUL7 as a biomarker in the prognosis evaluation of colon adenocarcinoma. Background Art

[0002] Colorectal cancer (CRC) is a common digestive tract tumor, the second most common cancer, and the fourth most common cause of cancer-related death. Colon adenocarcinoma (COAD) is the most common pathological type of CRC, accounting for approximately 95%. With changes in people's dietary patterns, the incidence of COAD has increased annually. In recent years, researchers have made significant progress in the diagnosis and treatment of COAD, but for COAD patients, distant metastasis remains the main cause of death. Literature reports that 70% of patients who die from COAD have distant metastases.

[0003] Therefore, it is necessary to identify new markers to further elucidate the biological characteristics of COAD, disclose the risk of COAD metastasis early, and predict clinical outcomes.

[0004] Tumor metastasis is a complex process involving the matrix, cell migration, and invasion. CUL7 (Cullin7), a relatively recent member of the Cullin protein family, is highly conserved and not itself an oncogene or tumor suppressor. It primarily influences cell signaling pathways through substrate proteins, further inducing tumor formation. Studies have shown that CUL7 is highly expressed in tumor cells, including choriocarcinoma, hepatocarcinoma, and breast cancer, acting as an oncogene and implicated in tumor proliferation, apoptosis, and malignant progression. Drugs and gene therapies targeting CUL7 are also gaining increasing attention. CUL7 overexpression plays a crucial role in the pathogenesis and progression of hepatocellular carcinoma and may be a key marker for its management. Silencing CUL7 significantly reduces the migration, invasion, and metastatic capacity of hepatocellular carcinoma cells. Furthermore, studies have found that overexpression of CUL7 is closely associated with poor prognosis in patients with epithelial ovarian cancer and can downregulate p53 expression, promoting ovarian cancer cell proliferation and invasion. Men et al. found that CUL7 overexpression in lung cancer is essential for lung cancer cell proliferation. Summary of the Invention

[0005] The first aspect of the present invention aims to provide the use of CUL7 as a biomarker.

[0006] The second aspect of the present invention aims to provide use of a substance for detecting CUL7 in preparing a product.

[0007] The third aspect of the present invention aims to provide the use of CUL7 as a target in screening drugs for treating or preventing colon cancer.

[0008] In order to achieve the above object, the technical solution adopted by the present invention is:

[0009] The first aspect of the present invention provides a use of CUL7 as a biomarker, wherein the use comprises at least one of (1) to (5);

[0010] (1) preparing a product for evaluating or predicting the prognostic risk of colon cancer;

[0011] (2) Prepare products for predicting the suitability of immunotherapy for colon cancer;

[0012] (3) preparing a product for predicting the survival rate of patients with colon cancer;

[0013] (4) Preparation of products for early screening of colon cancer;

[0014] (5) Preparation of products for disclosing the risk of colon cancer metastasis.

[0015] Preferably, the colon cancer comprises colon adenocarcinoma.

[0016] A second aspect of the present invention provides a use of detecting CUL7 substance in any one of (6) to (11);

[0017] (6) preparing products for evaluating or predicting the prognostic risk of colon cancer;

[0018] (7) Prepare products for predicting the suitability of immunotherapy for colon cancer;

[0019] (8) preparing a product for predicting the survival rate of patients with colon cancer;

[0020] (9) Develop treatment / medication plans for patients with gastrointestinal cancer;

[0021] (10) Preparation of products for early screening of colon cancer;

[0022] (11) Preparation of a product for disclosing the risk of colon cancer metastasis.

[0023] Preferably, the colon cancer comprises colon adenocarcinoma.

[0024] Preferably, the substance for detecting CUL7 comprises a substance for quantitatively detecting CUL7.

[0025] Preferably, the substance for detecting CUL7 includes a substance for detecting CUL7 at the gene and / or protein level.

[0026] Preferably, the substance includes a substance for detecting the expression level of CUL7 by enzyme-linked immunosorbent assay, immunofluorescence assay, radioimmunoassay, immunoprecipitation, immunoblotting, high performance liquid chromatography, capillary gel electrophoresis, near-infrared spectroscopy, mass spectrometry, immunochemiluminescence, colloidal gold immunoassay, fluorescence immunochromatography, surface plasmon resonance, immuno-PCR, biotin-avidin technique, PCR or biochip method.

[0027] Preferably, the substance for detecting CUL7 is selected from: a substance specific to CUL7, a CUL7-specific probe, a gene chip, and a PCR primer.

[0028] Preferably, the substance specific for CUL7 is any one of (b1) to (b3):

[0029] (b1) an antibody that specifically binds to CUL7;

[0030] (b2) a ligand protein or polypeptide that specifically binds to CUL7;

[0031] (b3) Non-protein compounds that specifically recognize CUL7.

[0032] Preferably, the antibody comprises at least one of a polyclonal antibody, a monoclonal antibody, a single-chain antibody, a functional antibody fragment, an antibody Fab region, a nanobody, a chimeric antibody, and a multispecific antibody.

[0033] Preferably, the products include but are not limited to reagents, kits, test papers, and chips.

[0034] Preferably, the test sample of the product is selected from at least one of body fluids, tissues, cells, and excretions of the subject to be tested.

[0035] The third aspect of the present invention provides the use of CUL7 as a target in screening drugs for treating or preventing colon cancer.

[0036] Preferably, the colon cancer comprises colon adenocarcinoma.

[0037] The beneficial effects of the present invention are:

[0038] This study used bioinformatics and tissue microarrays (TMA) to investigate the relationship between CUL7 expression and the prognosis and clinicopathological features of COAD. The results showed that CUL7 expression was significantly increased in COAD tumor tissue compared to normal tissue. The KM curve showed that high CUL7 expression in COAD was associated with decreased survival, suggesting that CUL7 could be used as a biomarker for early screening or prognosis of colon adenocarcinoma and as a therapeutic target in clinical trials. Furthermore, data were validated by collecting tissue samples from clinical patients and conducting in vitro experiments, confirming that CUL7 is highly expressed in COAD patients and that high CUL7 expression indicates a worse prognosis for COAD patients. High CUL7 expression could serve as an independent risk factor for assessing the recurrence risk of COAD. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 is the expression of CUL7 in COAD; wherein, a is the expression level of CUL7 in TCGA dataset, b is the expression level of CUL7 in TCGA and GTEx datasets, c is the paired expression level of CUL7 in cancer tissues and adjacent normal tissues in TCGA data, and d is the expression level of CUL7 in tumor cell lines in CCLE database. In the figure, * represents p < 0.05, ** represents p < 0.01, and *** represents p < 0.001.

[0040] Figure 2 Figure 3 is the correlation between CUL7 expression and overall survival (OS) in COAD; A is the Cox regression model analysis of the correlation between CUL7 expression and OS in COAD tumors; B is the Kaplan-Meier curve.

[0041] Figure 3 Figure 3 is the correlation between CUL7 expression and progression-free survival (PFI) in COAD; A is the Cox regression model analysis of the correlation between CUL7 expression and PFI in COAD tumors; B is the Kaplan-Meier curve.

[0042] Figure 4 Figure 3 is the correlation between CUL7 expression and disease-free survival (DFI) in COAD; A is the Cox regression model analysis of the correlation between CUL7 expression and DFI in COAD tumors; B is the Kaplan-Meier curve.

[0043] Figure 5 Figure 3 is the correlation between CUL7 expression and disease-specific survival (DSS) in COAD; A is the Cox regression model analysis of the correlation between CUL7 expression and DSS in COAD tumors; B is the Kaplan-Meier curve.

[0044] Figure 6is the correlation between CUL7 expression and immune-related biomarkers; A is the correlation between CUL7 expression and microsatellite instability (MSI), and B is the correlation between CUL7 expression and tumor mutation burden (TMB).

[0045] Figure 7 Figure 3 is the correlation between overexpression of CUL7 and the poor prognosis of COAD; A is the staining pattern of COAD and normal colon mucosa tissue microarrays, B is the CUL7 staining in the COAD cytoplasm, the left picture is the original picture, the right picture is the enlarged picture of the boxed part in the original picture, and the red arrows indicate positively stained cells, C is the expression result of CUL7 in the cytoplasm of COAD tissue, the left picture is the original picture, the right picture is the enlarged picture of the boxed part in the original picture, and the red arrows indicate positively stained cells, D is the expression result of CUL7 in normal colon mucosa tissue, the left picture is the original picture, and the right picture is the enlarged picture of the boxed part in the original picture, E is the immune response score result of COAD tissue, and F is the overall survival curve of COAD patients under different CUL7 expression conditions.

[0046] Figure 8 Figure 3 shows the biological effects of CUL7 overexpression on SW480 / SW620 cell lines; A is Western blot analysis of CUL7 expression in SW480 and SW620 cell lines after overexpression of CUL7; B is the growth curve of SW480 and SW620 cells after overexpression of CUL7; C is the apoptosis of SW480 and SW620 cells after overexpression of CUL7; D is Transwell analysis of the migration ability of SW480 and SW620 cells after overexpression of CUL7. In the figure, ** represents p < 0.01, and * represents p < 0.05.

[0047] Figure 9 Figure 3 shows the biological effects of CUL7 knockout on SW480 / SW620 cell lines; A is Western blot analysis of CUL7 expression in SW480 and SW620 cell lines after CUL7 knockout; B is the growth curve of SW480 and SW620 cells after CUL7 knockout; C is the apoptosis of SW480 and SW620 cells after CUL7 knockout; D is Transwell analysis of the migration ability of SW480 and SW620 cells after CUL7 knockout. In the figure, ** represents p < 0.01, and * represents p < 0.05. DETAILED DESCRIPTION

[0048] The present invention is further described in detail below through specific examples.

[0049] It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0050] Unless otherwise specified, the materials and reagents used in this example were obtained from commercial sources.

[0051] Example 1

[0052] The inventors used the TCGA database (https: / / portal.gdc.com), the Genotype Tissue Expression Database (GTEX; https: / / www.genome.gov / ), and the CCLE database to obtain CUL7 expression data between COAD tumor tissues and nearby normal tissues, and CUL7 expression data in normal human tissue samples, respectively.

[0053] Differential analysis using the TCGA database showed that the expression level of CUL7 in COAD tumor tissues was significantly increased compared with normal human tissues ( Figure 1 At the same time, the inventors downloaded the data of normal tissues from the GTEx database and analyzed them in combination with the TCGA dataset. They found that CUL7 was significantly expressed in COAD compared with normal tissues (p < 0.005) ( Figure 1 Further analysis of CUL7 expression levels in COAD tumor paired cancer tissues and adjacent normal tissues showed that CUL7 expression was higher in tumor tissues than in adjacent normal tissues ( Figure 1 In the CCLE database, the expression of CUL7 in COAD tumor cell lines ( Figure 1 In conclusion, CUL7 expression was significantly enhanced in COAD tumor tissues, suggesting that CUL7 may act as an oncogenic molecule and affect the development of COAD.

[0054] Example 2 Early diagnosis and prognostic evaluation of COAD by CUL7

[0055] Blood from COAD patients (100 patients pathologically diagnosed with COAD at Jiangmen Central Hospital from 2015 to 2016, all of whom signed informed consent forms) was collected for CUL7 expression level detection by ELISA to perform early diagnosis and prognosis assessment of COAD. The specific detection process is as follows: Serum acquisition: 5 mL of fasting venous blood was collected from the subjects, centrifuged at 3000 r / min for 30 minutes, and serum was separated; Coating: CUL7 monoclonal antibody was diluted to a protein content of 10 g / mL with 0.05 M pH 9 carbonate buffer, 0.1 mL was added to each reaction well of a 96-well polystyrene plate, and incubated at 4°C overnight; the next day, the solution in the well was discarded and the plate was analyzed using Thermo Scientific™ ELISA. Wash 3 times with washing buffer, 3 minutes each time; add sample: add 0.1 mL of diluted serum to be tested to the above-mentioned coated reaction wells, incubate at 37°C for 1 hour, and then wash (blank wells, negative control wells and positive control wells are also performed); add enzyme-labeled antibody: add 0.1 mL of freshly diluted enzyme-labeled antibody (1:100 diluted horseradish peroxidase-labeled streptavidin) to each reaction well, incubate at 37°C for 0.5 hour, discard the solution in the wells, and wash 3 times with washing buffer, 3 minutes each time; add substrate solution Color development: Add 0.1 mL of the temporarily prepared TMB substrate solution to each reaction well and incubate at 37°C for 30 minutes. Stop the reaction: Add 50 μL of stop solution (bicarbonate buffer) to each well to terminate the reaction and measure the experimental results within 20 minutes. Result evaluation: On the ELISA detector, at 450 nm (410 nm if developing with ABTS), adjust the absorbance (OD value) of each well to zero with the blank control well. If it is greater than twice the specified negative control OD value (0.05), it is positive.

[0056] A survival analysis of COAD patients was further performed using Kaplan-Meier curves. The relationship between CUL7 expression and prognosis was assessed using the following indicators: overall survival (OS), disease-specific survival (DSS), disease-free interval (DFI), and progression-free interval (PFI). Taking into account other factors such as tumors, which may lead to patient death during follow-up, the inventors calculated the hazard ratio (HR) and 95% confidence interval in the univariate regression survival analysis. After establishing the correlation between CUL7 expression and survival using the Kaplan-Meier plotter (http: / / kmplot.com / analysis / ), the hazard ratio (HR) with a 95% confidence interval and log-rank p-value was determined.

[0057] Overall survival (OS) results showed that CUL7 expression in COAD patients was significantly correlated with OS (p = 0.010), indicating that CUL7 expression is a strong high-risk predictor in COAD ( Figure 2Middle A). Kaplan-Meier analysis showed that higher CUL7 expression was associated with a worse survival probability in COAD patients (p=0.039). Figure 2 In the Cox regression analysis of progression-free survival (PFI), it was found that high expression of CUL7 was a risk factor for COAD (HR=1.631). Figure 3 Middle A), according to Kaplan-Meier analysis, high expression of CUL7 was associated with poor prognosis of COAD ( Figure 3 The DFI results showed that the inventors also found that high CUL7 expression in COAD was associated with low DFI (p < 0.001), with a hazard ratio (HR) of 3.842 ( Figure 4 Middle A), According to Kaplan-Meier analysis, high CUL7 expression was similar to low DFI in COAD patients (p=0.001) ( Figure 4 The disease-specific survival (DSS) of COAD was further evaluated. Cox regression analysis of DSS revealed that high CUL7 expression was a high-risk factor in COAD (p = 0.029), with a hazard ratio (HR) of 1.647 ( Figure 5 Middle A), According to Kaplan-Meier analysis, high expression of CUL7 in COAD patients was also associated with poor prognosis of COAD (p < 0.001) ( Figure 5 In conclusion, the KM curves showed that in COAD, high CUL7 expression was associated with decreased survival, especially poor disease-specific survival (DSS) and disease-free interval (DFI), suggesting that high CUL7 expression could be used as a marker for the prognosis of COAD patients.

[0058] Example 3 Correlation between CUL7 expression and microsatellite instability (MSI) or tumor mutational burden (TMB) in COAD

[0059] The TME plays a key role in cancer development and progression. Uncontrolled proliferation, resistance to apoptosis, and a metabolic shift toward anaerobic glycolysis are caused by genetic changes in tumor cells (Warburg effect). These processes lead to hypoxia, acidosis, and oxidative stress in the TME, initiating ECM regulation, triggering responses from surrounding immune cells (lymphocytes and macrophages) and stromal cells (e.g., fibroblasts), and ultimately contributing to cancer formation and metastasis. Therefore, revealing the COAD correlation between CUL7 expression and the TME is crucial.

[0060] The inventors analyzed the correlation between CUL7 expression and MSI / TMB in COAD tumors in TCGA. The MSI map results showed that CUL7 expression in COAD was negatively correlated with MSI ( Figure 6 TMB results showed that CUL7 expression in COAD was negatively correlated with TMB ( Figure 6 Middle B).

[0061] Example 4 Effect of high expression of CUL7 on the risk of invasion and metastasis in COAD

[0062] Furthermore, the inventors performed immunohistochemical staining analysis on COAD tissue and normal colon mucosa tissue using TMA tissue microarrays as follows: Immunohistochemistry (IHC) analysis of COAD tumor tissue was performed using Dako EnVision Systems (Dako Diagnostics, Switzerland) according to the manufacturer's instructions. Briefly, slides of TMA specimens were blocked by proteolysis and peroxidase digestion, then incubated with a rabbit anti-human UST polyclonal antibody (1:50 dilution) at 4°C overnight. The slides were then washed with PBS and incubated with a horseradish peroxidase-conjugated antibody (1:100 dilution) at room temperature for 30 minutes. Afterwards, the substrate chromogen was added for 1 minute to visualize protein staining. The stained TMA slides were analyzed using ImageScope v11 software. Protein expression levels were assessed semiquantitatively by two pathologists blinded to the patient information. The intensity of staining on each slide was graded as follows: none = 0, weak = 1, moderate = 2, and strong = 3. The positive percentage of each slide was also defined as follows: 0% to 25% = 1, >25% to 50% = 2, >50% to 75% = 3, and >75% to 100% = 4. Based on the intensity and percentage scores, the immunoreactivity score (IRS) was calculated, and all samples were divided into high UST expression (IRS > 4) and low UST expression (IRS ≤ 4).

[0063] The results showed that CUL7 was highly expressed in COAD tumor tissues ( Figure 7 Middle B), weakly positive expression in the cytoplasm of COAD tissue ( Figure 7 C), while it was negative in normal colon mucosal tissue ( Figure 7 The immune response scores of COAD tissues were higher than those of normal colon mucosa tissues ( Figure 7 Middle E), and high expression of CUL7 indicates a worse prognosis ( Figure 7 Middle F).

[0064] We further added the clinical data of COAD patients from Jiangmen Central Hospital to perform survival analysis and COX analysis, and found that high expression of CUL7 was an independent risk factor for invasion and metastasis in COAD patients, and indicated a worse prognosis (Tables 1 and 2).

[0065] Table 1 Correlation between CUL7 expression and clinicopathological features of COAD

[0066]

[0067]

[0068] Table 2 Prognostic value of CUL7 expression on overall survival in COAD patients

[0069]

[0070] Example 5 Biological effects of CUL7 overexpression / intervention on CCRC cells

[0071] The inventors further constructed PLKO.1-puro-CUL7 and PLKO.1-puro-shCUL7 lentiviral expression vectors, and used the lentiviral transfection method to construct SW480 / SW620 cell lines with stable overexpression of CUL7 (over-CUL7) and SW480 / SW620 cell lines with stable overexpression of CUL7 (i.e., knockout of CUL7, KO-CUL7). The specific operation is as follows: HEK293 cells were evenly seeded in a 10 cm dish (the dish was pre-coated with 0.1% gelatin), and transfection was performed when the cell density reached 70-80%; calcium phosphate method was used for transfection, and 2 μg of the target plasmid PLKO.1-puro-CUL7 / PLKO.1-puro-shCUL7 and its packaging plasmid psPAX2 plasmid and 2 μg of pMD2.G plasmid were shaken and added to 67 μL of calcium phosphate transfection reagent, mixed and allowed to stand at room temperature for 20 minutes; the transfection system was added to the dish, gently shaken to mix, and placed in an incubator for culture; after 6 hours, the medium was replaced with complete medium and cultured for another 48 hours; the culture medium was collected, centrifuged at 2000 rpm, and the supernatant was collected; ultrafiltration column was filtered, centrifuged at 6000 g for 30 minutes at 4°C to reduce the volume to 1 mL; the virus concentrate was divided and stored at -8 Store in a refrigerator at 0°C for future use; the target SW480 / SW620 cells were seeded into 6-well plates, cultured to a density of about 60%, washed once with physiological saline, added with 2 mL of complete culture medium, then added with 500 uL of virus concentrate, and added with polybrance reagent (to promote the fusion infection of virus particles) to a final concentration of 8 ug / mL, and cultured in an incubator for 6 hours; 2 mL of complete culture medium was added and the infection continued for 24 hours; after 24 hours, fresh complete culture medium was replaced and cultured for another 12 hours; 1 μg / mL puromycin was used to screen the positive clone cells that had been successfully transfected, amplified and verified, frozen, and stored for future use, thereby obtaining a CUL7 stably overexpressing SW480 / SW620 cell line (over-CUL7) and a CUL7 stably over-interference SW480 / SW620 cell line (KO-CUL7).

[0072] Western blot (see Wang C, Li X, et al. Front Oncol. 2021 Jul 28; 11: 656852) was further used to confirm the overexpression and interference effect of CUL7 in cells. CCK-8 (the adherent cells in the logarithmic phase were detached by adding trypsin; when most of the cells were detached, two times the volume of trypsin-containing medium was added to terminate the digestion; the cell suspension was transferred to a centrifuge tube, and the cell culture flask was rinsed with PBS buffer to rinse the remaining cells, and the PBS rinse was transferred to a centrifuge tube at 1000 r / min. The supernatant of the centrifuge tube was discarded and the tube was resuspended with 1 mL of serum-containing medium; the cells were counted using a counter and the number of diluted cells was maintained at 5 to 10 × 10 5 ; Set up a blank control in a 96-well plate, and set up 3 to 5 replicate wells for each group; add 100 μL of diluted cell suspension to each well; inoculate for 24 hours, and after the cells have well adhered to the wall, aspirate the culture medium from each well; add drug-free culture medium to the blank group, and add culture medium containing different concentrations of drugs to the experimental group; culture in an incubator for 24 hours, add culture medium containing CCK-8, and CCK-8 accounts for 1 / 10 of the volume of the cell culture medium; culture in an incubator for 1 hour, and when the color turns orange, it can be taken out for detection. The 96-well plate was taken out and placed in a microplate reader for detection with the wavelength set at 450 nm), flow cytometry (SW480 / SW620 cells with stable overexpression / interference of CUL7 were cultured for 48 hours, digested with trypsin, washed twice with cold PBS, centrifuged at 2000 rpm for 5 minutes, and the supernatant was discarded. The cells were resuspended in 0.3 mL of pre-cooled PBS containing 10% FBS and blown evenly, and then 0.7 mL of pre-cooled anhydrous ethanol was added and blown evenly, and incubated at 4°C overnight; the fixed cells were centrifuged at 3000 rpm for 3 minutes, the supernatant was discarded, and the cells were washed twice with cold PBS; the cells were resuspended in 0.1 mg / mL RNAase and incubated at 37°C for 30 minutes; and 0.1 mg / mL PI, incubated at 4°C with the light off for 20 min, and then flow cytometry analysis was performed), Transwell assay (Transwell assay was used to detect cell migration ability. The prepared stably overexpressed / interfered SW480 / SW620 cells were digested with Trypsin and counted, the culture medium was removed by centrifugation, and the cells were resuspended in 1640 medium without FBS to a density of 1×10 5 / mL; directly place the chamber into a 24-well plate, take 200 μL of the above cell suspension and add it to the upper layer of the chamber, add 600 μL of 1640 medium containing 10% FBS to the lower layer of the chamber in the 24-well plate, and culture in a 37°C incubator for 24 hours, then take out the chamber, carefully wipe the cells at the bottom of the upper layer of the chamber with a small cotton swab, fix the cells outside the chamber membrane with a mixture of methanol / glacial acetic acid (v / v=3 / 1), and culture at room temperature for 10-15 minutes; wash once with PBS, counterstain with hematoxylin for 10 minutes, rinse with tap water, observe under a microscope, take pictures, take pictures, and count the results) and cell scratch test (use a marker to draw horizontal lines on the back of the 6-well plate (with a ruler), cross at least 5 lines in each well, and each line should be even and parallel; inoculate about 5-10×10 5 The principle is that the cells can grow completely overnight, and it is important to spread them evenly. After 24 hours, use a 20uL pipette tip (sterilized) to scratch the black line on the back of the well plate vertically so that the scratch intersects with the marked line) to detect cell proliferation, apoptosis, migration and invasion abilities to clarify the biological effects of CUL7 overexpression / interference on CRC cells. CRC cells without overexpression or interference of CUL7 were used as controls (i.e., over-NC and KO-NC).

[0073] The results showed that Figure 8 and Figure 9 As shown in Figure 3, overexpression of CUL7 promoted the invasion and metastasis of CRC cells, increased their invasive ability, and further led to apoptosis of SW480 and SW620 cells ( Figure 8 Knockout of CUL7 inhibits CRC cell invasion and metastasis, and inhibits apoptosis of SW480 and SW620 cells ( Figure 9 ).

[0074] While the embodiments of the present invention have been described in detail above with reference to the accompanying drawings, the present invention is not limited to the embodiments described above. Various modifications may be made within the scope of knowledge possessed by a person skilled in the art without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof may be combined with one another unless there is a conflict.

Claims

1. Application of substances used to detect CUL7 in (1) or (2); (1) Preparing a product for predicting the prognostic risk of colon adenocarcinoma; (2) Prepare a product for predicting the survival rate of patients with colon adenocarcinoma.

2. The use according to claim 1, characterized in that The substance for detecting CUL7 includes a substance for quantitatively detecting CUL7.

3. The use according to claim 2, characterized in that The substances include substances for detecting CUL7 at the gene or protein level.

4. The use according to claim 3, characterized in that The substance for detecting CUL7 includes a substance for detecting CUL7 by enzyme-linked immunosorbent assay, immunofluorescence, radioimmunoassay, immunoprecipitation, immunoblotting, high performance liquid chromatography, capillary gel electrophoresis, near-infrared spectroscopy, mass spectrometry, immunochemiluminescence, colloidal gold immunoassay, fluorescent immunochromatography, surface plasmon resonance, immuno-PCR or biotin-avidin technology.

5. The use according to any one of claims 1 to 4, characterized in that The product comprises at least one of a reagent, a test kit, a test paper, and a chip.

6. The use according to any one of claims 1 to 4, characterized in that The test sample of the product is selected from at least one of the body fluids, tissues, cells, and excretions of the subject to be tested.

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