CN111321221A - Compositions, microarrays and computer systems for predicting recurrence risk after local resection of rectal cancer - Google Patents

Abstract

The invention discloses a composition, a microarray and a computer system for predicting the risk of recurrence after a local resection surgery for rectal cancer. The invention adopts a microarray chip mode, screens out specific rectal cancer postoperative recurrence marker genes aiming at Chinese population, draws out a specific rectal cancer postoperative recurrence evaluation model, customizes the rectal cancer postoperative recurrence marker gene microarray chip according to research results, and is used for clinical diagnosis and evaluation of rectal cancer patients postoperative recurrence evaluation of Chinese population.

Description

Composition, microarray for predicting the risk of recurrence after local resection of rectal cancer and computer systems

technical field

The invention relates to the field of biomedicine, in particular to a composition, a microarray and a computer system for predicting the risk of recurrence after local excision of rectal cancer.

Background technique

Colorectal cancer (CRC) is a malignant tumor originating from the colorectal mucosal epithelium. It is the third most common malignant tumor in the world and the fourth most common cause of cancer mortality in the world. In my country, there are more than 250,000 new cases of colorectal cancer and about 140,000 deaths each year. Both new cases and deaths account for 20% of the world's colorectal cancer cases during the same period. Given the low position of the rectum, its removable precancerous lesions are incomplete and the risk of disease recurrence is extremely high. Therefore, establishing an effective rectal cancer risk assessment method to reduce the morbidity and mortality of rectal cancer in my country is a major clinical problem that cannot be solved without delay.

In patients with early-stage colorectal cancer, surgical resection is usually performed to effectively control the further development and spread of the tumor. Colorectal cancer often recurs due to hematogenous metastasis two years after surgery, and postoperative adjuvant chemotherapy aimed at eliminating minimal residual disease in the body can reduce the recurrence rate and improve the long-term survival rate of patients. Therefore, postoperative patients need to pay close attention to tumor metastasis and recurrence, and take effective diagnosis and treatment plans in time to prevent disease progression. Common diagnostic methods for rectal cancer include: fecal occult blood test, digital rectal examination, proctoscopy, sigmoidoscopy, colonoscopy, imaging examination and the detection of CEA carcinoembryonic antigen. These screening methods have their own limitations, such as low detection sensitivity or not easy to detect frequently. In view of the rapid development of current high-throughput detection technology, research on blood biomarker-based detection is also emerging. First, extracorporeal blood tests are safe and minimally invasive. Second, no dietary restrictions, colon cleansing or sedation are required. Third, sample collection and processing procedures may be easier and more convenient. Furthermore, there is no microbial community that could degrade biomarkers or hinder analysis. Therefore, the risk of recurrence of rectal cancer can be predicted in time through peripheral blood detection, and the accuracy of detection can be improved, thereby saving lives.

For the screening and diagnosis of colorectal cancer, the most studied is the study of septin9 methylation. Studies have confirmed that the sensitivity of septin9 for screening colorectal cancer is about 70%, and the specificity is about 90%. However, DNA microarray technology can simultaneously quantify the expression of thousands of genes and can better probe the complex biological mechanisms that lead to rectal tumorigenesis and progression than individual gene markers. There are numerous reports using DNA microarray technology to identify blood-based gene expression signatures for CRC detection. Han and colleagues reported a 5-gene expression signature in colorectal cancer with a detection sensitivity of 88% and a specificity of 64%; Marshall et al. published a 7-gene expression signature in colorectal cancer with a detection sensitivity of 72% and a specificity of 64%. 70%. Rosenthal et al also reported a panel of 202 colorectal cancer expression-related genes with a detection sensitivity of 90% and specificity of 88%.

However, the current research reports on colorectal cancer are mainly based on data from European and American populations, and the reported data on Asian populations or Chinese populations are not detailed.

SUMMARY OF THE INVENTION

In view of the above situation, the present invention conducts screening of relevant rectal cancer postoperative recurrence marker genes for rectal cancer patients in the Chinese population, establishes a corresponding postoperative recurrence evaluation model, and further customizes a specific postoperative recurrence detection method for the Chinese rectal cancer patient population. Program. The present invention has been completed based on at least this. Specifically, the present invention includes the following.

In a first aspect of the present invention, there is provided a composition for predicting the risk of recurrence after local resection of rectal cancer, comprising a composition that specifically binds to a compound selected from the group consisting of WDR43, RAB31, FASN, TYMS, SNX2, DAP3, MRPS18C, FANCL, CIDECP Oligonucleotides of partially contiguous sequences of genes in the group consisting of , HNRNPH1 and CMSS1.

In certain embodiments, the contiguous sequence of the present invention is a conserved sequence of the corresponding gene.

In certain embodiments, the oligonucleotides of the present invention are designed to be capable of immobilizing one end of the oligonucleotide to bind to a substrate, and at least a portion of the sequence can specifically bind to a sequence selected from the group consisting of WDR43, RAB31, FASN, TYMS, SNX2, DAP3 Partially contiguous sequences of genes in the group consisting of , MRPS18C, FANCL, CIDECP, HNRNPH1, and CMSS1, enabling the capture of the desired gene in the sample.

In a second aspect of the present invention, there is provided a microarray for predicting the risk of recurrence after local excision of rectal cancer, which comprises the composition of the present invention and a substrate, the surface of the substrate has a plurality of site.

Preferably, each site has a reactive functional group capable of binding to the oligonucleotide; or each site binds to one of the oligonucleotides, respectively.

Preferably, each site is regularly arranged in such a manner that the interval between adjacent sites is in the range of 0.1-20 μm, and the area of each site is less than 1 μm ² .

Preferably, the oligonucleotide comprises repeating sequences of multiple target regions, and the sequence length of the oligonucleotide is 50-500 nt.

A third aspect of the present invention provides a computer system for predicting the risk of recurrence after local resection of rectal cancer, comprising:

a. An input device for receiving subject data, wherein the subject data includes genetic testing data from the microarray of the present invention;

b. a memory with a database for storing at least the genetic testing data;

c. A processor in communication with the memory and configured to:

Calculate an evaluation value F(x) through an algorithmic model using the genetic testing data; and

d. An output device configured to send a notification according to the result of the judgment of the evaluation value F(x).

Preferably, the algorithm model is:

F(x)=29.3341-0.7416*WDR43+0.9450*RAB31+2.9083*FASN-0.5904*TYMS-3.0449*SNX2-2.5533*DAP3+3.7309*pt+1.9524*MRPS18C+1.0107*FANCL-6.5343*NPCIDECP9*1.19 +0.1941*CMSS1.

Preferably, the processor is further configured to: obtain the P(x) value by the following formula, and compare the obtained P(x) value with a reference value of 0.37, and determine the local rectal cancer when P(x) is higher than 0.37 The risk of recurrence after resection was high, and the risk of recurrence after local resection of rectal cancer was determined to be low when P(x) was less than or equal to 0.37.

The composition, the microarray and the computer system of the present invention are developed for the specific postoperative recurrence of the Chinese rectal cancer patient population, and can be used for model validation of the clinical samples of the Chinese rectal cancer patient population. The results confirmed that the microarray for postoperative recurrence of rectal cancer of the present invention combined with the postoperative recurrence assessment model to evaluate postoperative recurrence of rectal cancer, the sensitivity of postoperative recurrence assessment was 78%, and the specificity was 100%.

Description of drawings

Figure 1. The results of random forest analysis.

Detailed ways

Various exemplary embodiments of the present invention will now be described in detail, which detailed description should not be construed as a limitation of the invention, but rather as a more detailed description of certain aspects, features, and embodiments of the invention.

It should be understood that the terms described in the present invention are only used to describe particular embodiments, and are not used to limit the present invention. Additionally, for numerical ranges in the present disclosure, it should be understood that the upper and lower limits of the range, and every intervening value therebetween, are specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated value or intervening value in that stated range is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention relates. Although only the preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials in connection with which the documents are referred. In the event of conflict with any incorporated document, the content of this specification controls. "%" is a percentage by weight unless otherwise stated.

The present invention is completed on the basis of the results of in-depth research on the Chinese rectal cancer patient population. In the study, strict screening was carried out for the enrolled population. All enrolled populations were patients with rectal cancer after stage II. The experimental group consisted of known postoperative recurrences, and the control group consisted of patients who had basically no postoperative recurrence. Patients with signs of recurrence. The experimental group consisted of blood samples from 68 patients with postoperative recurrence of stage II rectal cancer, and the control group consisted of blood samples from 68 patients with no recurrence of rectal cancer after surgery. RNA was extracted and detected by microarray chip (Affymetrix Gene Chips HG-U133Plus 2.0). 500 genes related to the pathogenesis of rectal cancer were selected for random forest analysis and a postoperative recurrence assessment model was established; the accuracy of the model was verified by leave-one-out cross-validation (LOOCV) method, and finally 11 genes were found to be associated with postoperative recurrence and postoperative recurrence. There is a certain difference in expression between rectal cancer and non-recurrence of rectal cancer after surgery; 9 patients in the experimental group and the control group were used to do a blind test to test the applicability of the data model and the accuracy of the test. Finally, the technical solution of the present invention is customized.

[Composition for predicting recurrence risk after local excision of rectal cancer]

In a first aspect of the present invention, there is provided a composition for predicting the risk of recurrence after local resection of rectal cancer, comprising a composition that specifically binds to a compound selected from the group consisting of WDR43, RAB31, FASN, TYMS, SNX2, DAP3, MRPS18C, FANCL, CIDECP Oligonucleotides of partially contiguous sequences of genes in the group consisting of , HNRNPH1 and CMSS1.

In the composition of the present invention, WDR43, RAB31, FASN, TYMS, SNX2, DAP3, MRPS18C, FANCL, CIDECP, HNRNPH1 and CMSS1 are respectively referred to as target genes. The invention can predict the recurrence risk of rectal cancer after local excision operation through a smaller number of target genes, and the effect is better. The present invention can use at least one of the above-mentioned target genes. In order to improve the accuracy of the prediction results, the present invention preferably uses a plurality of target genes, and more preferably uses all the above-mentioned target genes.

In general, there are at least one, preferably multiple, oligonucleotides that specifically bind to the same target gene. For example, the oligonucleotides that specifically bind to the target gene WDR43 may be multiple oligonucleotides that specifically bind to different contiguous sequences of the gene. The contiguous sequence here is preferably the conserved sequence of the gene. Conserved sequences for these genes are known in the art and can be readily determined with general knowledge.

In certain embodiments, the oligonucleotides of the present invention are designed to be capable of immobilizing one end of the oligonucleotide to bind to a substrate, and at least a portion of the sequence can specifically bind to a sequence selected from the group consisting of WDR43, RAB31, FASN, TYMS, SNX2, DAP3 Partially contiguous sequences of genes in the group consisting of , MRPS18C, FANCL, CIDECP, HNRNPH1, and CMSS1, enabling the capture of the desired gene in the sample. At this point the oligonucleotides are used to capture the target gene. The sequence length of such oligonucleotides is generally 50-500 nt, eg, 60-400 nt, 70-300 nt. Also preferably, the oligonucleotides of the invention comprise multiple (eg, 5-15) repeats of the target region, more preferably, linker sequences between the repeats. In a certain embodiment, the oligonucleotide comprises 10 repeats with a linker sequence of 5-50 nucleotides in length between adjacent repeats.

[Microarray for predicting recurrence risk after local excision of rectal cancer]

The second aspect of the present invention provides a microarray for predicting the risk of recurrence after local excision of rectal cancer, which can also be called "gene chip" or "microarray chip", which is a miniaturized, high-throughput The main principle of the gene detection and analysis technology is to hybridize the oligonucleotide or probe on the surface of the solid support with the fluorescently pre-labeled sample nucleic acid, and obtain the detection result of the sample by detecting and analyzing the hybridization signal.

The microarray of the present invention includes the composition and the substrate described in the first aspect. The composition has been described in detail above, and will not be repeated here. The base material will be described below.

The base material of the present invention is preferably formed of a generally used material and preferably has a plate shape. There are multiple independently arranged sites on the surface of the substrate. In certain embodiments, each site each has a reactive functional group, such as an aldehyde group, capable of binding to an oligonucleotide. In certain embodiments, each site of the present invention binds to one of the oligonucleotides described in the present invention, more preferably, one site binds to one oligonucleotide. In the microarray of the present invention, each site is regularly arranged in such a manner that the interval between adjacent sites is in the range of 0.1-20 μm, and the area of each site is less than 1 μm ² .

The preparation of the microarrays of the present invention can be carried out using known methods. In an exemplary method, it includes designing and synthesizing specific probes for related genes, and spraying the dissolved and diluted probes on a well-known commercially available aldehyde substrate by a gene chip spotter to solidify them for rectal application. Microarrays for recurrence risk after local excision of cancer.

The microarray of the present invention has great advantages in many aspects such as the speed of gene locus detection, the reliability of the detection of medium and low expression levels of target genes, etc., and has a complete and mature quality control process and analysis means, which is fast and convenient And has a high degree of accuracy and sensitivity. Therefore, the risk prediction method for recurrence of rectal cancer after local excision of rectal cancer based on microarray detection technology has a good clinical application prospect.

When using a microarray for predicting the risk of recurrence after local excision of rectal cancer, the following steps are generally included: (1) extracting the total RNA of the sample to be tested, and reverse transcribing it into the product to be hybridized; (2) in a suitable reaction The step of reacting the product to be hybridized with the microarray under the conditions; (3) the step of detecting the hybridization signal to obtain gene detection data.

[computer system]

A fourth aspect of the present invention provides a computer system for predicting the risk of recurrence after local resection of rectal cancer, comprising:

a. An input device for receiving subject data, wherein the subject data includes genetic testing data from the microarray of the present invention;

b. a memory with a database for storing at least the genetic testing data;

c. A processor in communication with the memory and configured to:

Calculate an evaluation value F(x) through an algorithmic model using the genetic testing data; and

d. An output device configured to send a notification according to the result of the judgment of the evaluation value F(x).

In the computer system of the present invention, preferably, the algorithm model is:

F(x)=29.3341-0.7416*WDR43+0.9450*RAB31+2.9083*FASN-0.5904*TYMS-3.0449*SNX2-2.5533*DAP3+3.7309*pt+1.9524*MRPS18C+1.0107*FANCL-6.5343*NPCIDECP9*1.19 +0.1941*CMSS1.

Further, according to the P(x) value obtained by the following formula, when P(x) is higher than 0.37, it is determined that the risk of recurrence after local resection of rectal cancer is high, and when P(x) is lower than or equal to 0.37, it is determined that local resection of rectal cancer is performed. The risk of later recurrence is low.

In the computer system of the present invention, the input device for receiving object data includes any form of input device. In the present invention, the object data includes at least the genetic detection data obtained from the microarray of the present invention or based on the microarray. These data include relative expression levels of each target gene, etc.

In the computer system of the present invention, a memory having a database is used to store at least the above-mentioned genetic testing data. The memory itself is a commonly used product in the field. Preferably, the memory is in communication with the input device, the output device or the processor to enable efficient exchange of data between the different components. In certain embodiments, the database in memory may be one or more. In the case of multiple databases, the data of each database can interact or exchange, or can interact or communicate with the processor separately, so as to ensure the effective performance of the evaluation. Preferably, the memory has a data management component, so as to effectively manage various types of data and improve data utilization efficiency.

In the computer system of the present invention, the processor is in communication with at least the memory, and is configured to calculate the sensitivity index through the algorithm model using the genetic testing data. Different algorithm models are extremely important to achieve the purpose of the present invention. Different algorithms get different results, sometimes not even at all. On the basis of a large number of in-depth studies, the present invention finds that the evaluation value obtained by the above algorithm has unexpected sensitivity and accuracy in predicting the risk of recurrence after local excision of rectal cancer.

Example 1

1. Collection of samples

Each patient uses PAXgene special RNA blood sample collection tube to collect blood samples. RNA extraction is performed immediately after sample collection to ensure that RNA is not degraded and to ensure sample quality.

2. RNA extraction

Blood RNA管)。严格按照试剂盒使用说明书进行操作；1. For each patient's blood sample, take 2.5mL of peripheral blood sample for RNA extraction. The extraction kit is MagMAX ^TM (with

Blood RNA tubes). Strictly follow the kit instructions for operation;

2. RNA quality identification: use Nanodrop OD 260nm to identify RNA purity, requiring OD260/280=2.0～2.2;

3. RNA quantification: RNA quantification was performed on the Agilent 2100 Bioanalyzer detection instrument using the RNA 6000 Nano Lab Chip kit. The entire operation process was completely operated in accordance with the kit's operating instructions.

3. Microarray chip hybridization

1. RNA reverse transcription: 50ng RNA was reverse transcribed using a reverse transcription kit (SuperScript ^™ IV ReverseTranscriptase, Invitrogen) to obtain cDNA, and the entire process was completely operated in accordance with the kit instructions;

2. cDNA purification: The reverse transcription product was purified by QIAquick PCR purification kit.

3. Sonication of cDNA: Adjust the concentration of cDNA to 10ng/μL, and use the Covaris Sonicator to perform sonication according to this procedure: 150bp, 340s. Peak Power 75, Duty Factor10, Cycle 200, Setpoint 20°C. The whole process is completely in accordance with the instruction manual of the Covaris ultrasonic interrupter.

4. Preparation of cDNA before hybridization Hybridization with microarray chip: Biotin labeling of cDNA before hybridization is performed according to the instructions of the microarray chip (GeneChip U133plus2), and chip hybridization is performed in strict accordance with the operating specifications of the instructions.

5. Observation of results: After the hybridization of the microarray chip, GeneChip Scanner3000 was used to observe the results.

Fourth, the data analysis process

1. Sample assignment: digitally convert the clinical information of each patient. The conversion relationship between patient clinical information and numbers is shown in the following table.

Table 1 - Correspondence table of clinical information and numerical conversion

clinical information define value relapse 1 no relapse 0 distal 1 proximal 0 female 1 male 0 >45 1 <=45 0

2. Data analysis: 500 genes related to the pathogenesis of rectal cancer were selected, and the expression results of these 500 genes were analyzed by random forest method. The analysis results are shown in Figure 1. The results showed that 11 genes (WDR43, RAB31, FASN, TYMS, SNX2, DAP3, MRPS18C, FANCL, CIDECP, HNRNPH1, CMSS1) showed different expression patterns in different patients in the experimental group and the control group.

3. Establishment of risk assessment equation: According to the analysis results of random forest method, establish postoperative recurrence assessment model: F(x)=29.3341-0.7416*WDR43+0.9450*RAB31+2.9083*FASN-0.5904*TYMS-3.0449*SNX2-2.5533 *DAP3+3.7309*pt+1.9524*MRPS18C+1.0107*FANCL-6.5343*CIDECP-1.1939*HNRNPH1+0.1941*CMSS1.

4. Cutoff value setting: P(x)=1/(1+exp(-F(x))), comprehensively analyze the data results of 53 samples, set an appropriate cutoff value, P(x)>0.37 is considered a surgical procedure Posterior rectal cancer has a higher risk of recurrence.

5. 5 patients with postoperative recurrence (samples 6-10) and 5 patients with no signs of recurrence after surgery (samples 1-5) were used for blind testing to test the applicability of the data model and the accuracy of the test. The results are shown in Table 1.

Table 1

Example 2

Probes were designed for each of the 11 genes in Example 1, the sequences of which are shown in SEQ ID NOs: 1-33, and the dissolved and diluted probes were sprayed and solidified on well-known commercially available probes by a gene chip spotter. On aldehydeylated substrates, microarrays were prepared for the risk of recurrence after local resection of rectal cancer. The specific preparation of the microarray was completed by Boao Biological Co., Ltd.

Table 2 - Probe Information

In this example, 18 clinical samples were used to test the applicability of the microarray chip. The specific results are shown in Table 3 and Table 4.

Table 3-11-gene microarray application results of 9 cases of rectal cancer that did not recur after surgery

gene name Sample 1 Sample 2 Sample 3 Sample 4 Sample 5 Sample 6 Sample 7 Sample 8 Sample 9 HNRNPH1 6.26 5.56 5.86 6.36 6.50 7.74 6.41 6.66 6.74 SNX2 6.86 7.04 7.20 6.85 7.22 7.41 7.03 7.33 7.36 FASN 5.54 6.24 6.86 6.15 6.31 6.75 6.25 6.05 5.66 WDR43 6.72 6.65 6.82 6.58 6.79 7.21 6.69 7.00 6.64 DAP3 8.24 8.44 8.58 8.26 8.98 8.82 8.56 8.92 9.06 RAB31 9.28 8.76 8.51 6.61 7.47 7.30 8.16 7.50 7.62 MRPS18C 3.44 3.92 3.82 4.04 4.27 3.96 3.76 4.39 4.21 FANCL 6.23 6.22 6.38 7.07 7.29 6.99 7.06 7.16 7.61 CMSS1 7.23 7.54 7.29 7.42 7.83 7.36 8.01 8.44 7.21 TYMS 6.54 6.01 5.88 6.95 7.60 8.81 5.88 7.04 6.41 CIDECP 3.76 3.91 3.89 4.06 4.15 3.96 3.89 3.99 4.11 pt 3 3 3 3 3 4 3 3 3 F(x) -2.986 -1.288 -0.944 -3.877 -6.144 -3.767 -2.351 -5.844 -7.666 P value 0.048 0.216 0.280 0.020 0.002 0.023 0.087 0.003 0.000 Predict N N N N N N N N N Clinical N N N N N N N N N

Table 4-9 Application results of 11-gene microarray chip in 9 cases of postoperative recurrence of rectal cancer

It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present invention without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the present invention. The description and examples of the present application are only exemplary.

SEQUENCE LISTING

<110> Cancer Hospital, Chinese Academy of Medical Sciences

Yuancode Gene Technology (Beijing) Co., Ltd.

<120> Compositions, microarrays and computer systems for predicting recurrence risk after local resection of rectal cancer

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<213> Artificial sequences

<400> 29

tgctaaagag ctgtcttcca agggagtgaa aatctgggat gccaatggat cccgagactt 60

tttggacagc ctgg 74

<210> 30

<211> 76

<212> DNA

<213> Artificial sequences

<400> 30

cttgggccca gtttatggct tccagtggag gcattttggg gcagaataca gagatatgga 60

atcagattat tcagga 76

<210> 31

<211> 70

<212> DNA

<213> Artificial sequences

<400> 31

cccatcgtta tcactcttcg tagacatgat ccgccactac gtgtccatcc tgctggagag 60

cgacaagaag 70

<210> 32

<211> 70

<212> DNA

<213> Artificial sequences

<400> 32

ctcacccagg aacaagtatc tgacagggga cgaggcaccc acagtccctc tcccataagc 60

ctgccaagaa 70

<210> 33

<211> 71

<212> DNA

<213> Artificial sequences

<400> 33

gattgatgtg gcccgtgtaa cctttgacct gtacaagctg aacccacagg acttcattgg 60

ctgcctgaac a 71

Claims (10)

1. A composition for predicting the risk of recurrence after local resection of rectal cancer, comprising specifically binding to a compound selected from the group consisting of WDR43, RAB31, FASN, TYMS, SNX2, DAP3, MRPS18C, FANCL, CIDECP, HNRNPH1 and CMSS1 Oligonucleotides of partially contiguous sequences of genes in a set. 2 . The composition for predicting the risk of recurrence after local excision of rectal cancer according to claim 1 , wherein the continuous sequence is a conserved sequence of the corresponding gene. 3 . 3. The composition for predicting the risk of recurrence after local excision of rectal cancer according to claim 1, wherein the oligonucleotide is designed so that one end thereof can be fixedly bound to a substrate, and at least part of the sequence can be Specifically binds to a partially contiguous sequence of genes selected from the group consisting of WDR43, RAB31, FASN, TYMS, SNX2, DAP3, MRPS18C, FANCL, CIDECP, HNRNPH1 and CMSS1, thereby enabling capture of the desired gene in the sample. 4. A microarray for predicting the risk of recurrence after local excision of rectal cancer, comprising the composition according to any one of claims 1-3 and a substrate, the surface of the substrate having a plurality of mutually independent Set multiple sites. 5. The microarray for predicting the risk of recurrence after local resection of rectal cancer according to claim 4, wherein each site has a reactive functional group capable of binding to the oligonucleotide; or each site binds the One of the oligonucleotides. 6. The microarray for predicting the risk of recurrence after local excision of rectal cancer according to claim 4 or 5, wherein each site is regularly arranged in a manner that the interval between adjacent sites is in the range of 0.1-20 μm, and each site is arranged regularly. The area of the dots is less than 1 μm ² . 7. The microarray for predicting the risk of recurrence after local resection of rectal cancer according to claim 4 or 5, wherein the oligonucleotide comprises repeating sequences of a plurality of target regions, and the oligonucleotide has The sequence length is 50-500nt. 8. A computer system for predicting the risk of recurrence after local resection of rectal cancer, comprising: a. An input device for receiving subject data, wherein the subject data comprises genetic detection data from the microarray of any one of claims 4-7; b. a memory with a database for storing at least the genetic testing data; c. A processor in communication with the memory and configured to: Calculate the evaluation value F(x) through an algorithmic model using the genetic testing data; and d. An output device configured to send a notification according to the result of the judgment of the evaluation value F(x). 9. The computer system for predicting recurrence risk after local resection of rectal cancer according to claim 8, wherein the algorithm model is: F(x)=29.3341-0.7416*WDR43+0.9450*RAB31+2.9083*FASN-0.5904*TYMS-3.0449*SNX2-2.5533*DAP3+3.7309*pt+1.9524*MRPS18C+1.0107*FANCL-6.5343*NPCIDECP9*1.19 +0.1941*CMSS1. 10. The computer system for predicting the risk of recurrence after local resection of rectal cancer according to claim 9, wherein the processor is further configured to: obtain the P(x) value by the following formula, and use the obtained P(x) ) value was compared with the reference value of 0.37. When P(x) was higher than 0.37, the risk of recurrence after local resection of rectal cancer was determined to be high, and when P(x) was lower than or equal to 0.37, the risk of recurrence of rectal cancer after local resection was determined to be low.

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