Detection marker and detection device for identifying primary and intra-wall metastasis of esophageal multifocal squamous carcinoma

Abstract

The invention relates to a detection marker and a detection device for identifying primary and intrawall metastasis of esophageal multifocal squamous carcinoma, and belongs to the technical field of cancer detection markers. According to the invention, a detection site set composed of 20 genes is constructed, and the gene diagnosis purpose of distinguishing primary and intrawall metastasis of esophageal multi-lesion squamous cell carcinoma is realized by analyzing the difference of mutation types on lesions.

Description

Technical Field

[0001] The invention relates to a detection marker and a detection device for distinguishing primary and intramural metastasis of esophageal multifocal squamous cell carcinoma, and belongs to the technical field of cancer detection markers. Background Art

[0002] Esophageal multifocal squamous cell carcinoma can be divided into multiple primary squamous cell carcinoma and intramural metastatic squamous cell carcinoma. However, it is often difficult to accurately distinguish multiple primary esophageal squamous cell carcinoma from intramural metastatic squamous cell carcinoma based on tissue pathology alone. It is hoped that a kit that can distinguish multiple primary esophageal squamous cell carcinoma from intramural metastatic squamous cell carcinoma at the molecular level can be constructed for the differential diagnosis of clinical multifocal esophageal squamous cell carcinoma. In the early stage, WES was performed on 14 samples of esophageal multifocal squamous cell carcinoma. The results showed that the use of WES can play an auxiliary diagnostic role in distinguishing multiple primary esophageal squamous cell carcinoma from intramural metastatic squamous cell carcinoma. However, WES is complicated to operate and is not suitable for clinical molecular diagnosis.

[0003] References: Li W, Cheng N, Zhao Z, et al. Molecular characteristics of multifocal esophageal squamous cell carcinomas to discriminate multicentric origin from intraramural metastasis[J]. The Journal of Pathology, 2022, 258(4): 395-407. Summary of the invention

[0004] The present invention provides a marker for distinguishing primary and intramural metastasis of esophageal multifocal squamous cell carcinoma based on gene molecular diagnosis.

[0005] The specific technical solution is:

[0006] The invention relates to an application of a mutation detection reagent of a gene marker in the preparation of a diagnostic reagent for distinguishing primary and intramural metastasis of multifocal squamous cell carcinoma of the esophagus, wherein the gene marker is composed of the following 20 genes: TP53, TTN, NOTCH1, PCLO, DAMTSL3, ZNF750, DNAH3, EP300, CDKN2A, MBTPS1, MYH15, RHBG, RLF, SYNE1, XIRP2, C6orf118, EYS, FILIP1L, LTBP1, and NCOA2.

[0007] The mutation detection reagent is used to detect exon mutations of gene markers.

[0008] A detection device for distinguishing primary and intramural metastasis of multifocal squamous cell carcinoma of the esophagus, comprising:

[0009] Gene detection module, used to detect mutations in the following 20 genes: TP53, TTN, NOTCH1, PCLO, DAMTSL3, ZNF750, DNAH3, EP300, CDKN2A, MBTPS1, MYH15, RHBG, RLF, SYNE1, XIRP2, C6orf118, EYS, FILIP1L, LTBP1, NCOA2;

[0010] The comparison module compares the information obtained by the gene detection module on different esophageal cancer foci of the same person to be tested. If the types of gene mutations in different cancer foci are the same, it is intramural metastasis; if they are different, it is multiple primary cancers.

[0011] A computer-readable medium for distinguishing primary and intramural metastasis of multifocal squamous cell carcinoma of the esophagus, wherein the medium records a computer program capable of executing the following method:

[0012] Step 1, obtaining mutation information of gene markers on different cancer lesions of the same person to be tested, wherein the gene markers are composed of the following 20 genes: TP53, TTN, NOTCH1, PCLO, DAMTSL3, ZNF750, DNAH3, EP300, CDKN2A, MBTPS1, MYH15, RHBG, RLF, SYNE1, XIRP2, C6orf118, EYS, FILIP1L, LTBP1, NCOA2;

[0013] Step 2: Obtain the gene mutation spectra of different cancer foci. If they are the same, it is judged as intramural metastasis; if they are different, it is multiple primary cancers. DETAILED DESCRIPTION

[0014] The specific first set of sample information used in the technical research of this patent is:

[0015]

[0016]

[0017] Two different tumor areas from the same tumor were sampled and named Ta and Tb, respectively.

[0018] 20 high-frequency mutation genes and their detection rates determined based on previous WES results.

[0019] Gene Mutation frequency TP53 67％ TTN 27％ NOTCH1 23％ PCLO 22％ DAMTSL3 18％ ZNF750 18％ DNAH3 16％ EP300 16％ CDKN2A 15％ MBTPS1 14％ MYH15 14％ RHB 14％ RLF 14％ SYNE1 14％ XIRP2 14％ C6orf118 12％ EYS 12％ FILIP1L 12％ LTBP1 12％ NCOA2 12％

[0020] On this basis, we re-selected 8 patients with multifocal squamous cell carcinoma of the esophagus after surgery and whose DNA quality control was qualified. We extracted DNA from each cancer foci of each patient and tested it through our 20-gene panel. Based on its mutation analysis, we compared the results. If there are mutations at the same site between different cancer foci of the same patient, it indicates intramural metastatic squamous cell carcinoma. If there are mutations at different sites between different cancer foci of the same patient, it indicates multiple primary squamous cell carcinoma. This can be seen from our results.

[0021]

[0022]

[0023] A panel of 20 genes with high frequency mutations was formed. This panel was used to perform molecular detection of different foci of esophageal multifocal squamous cell carcinoma by next-generation sequencing. The similarities and differences of gene mutations in different foci were compared to differentiate between multiple primary squamous cell carcinoma of the esophagus and intramural metastatic squamous cell carcinoma. Patients with consistent mutation spectra can be considered as intramural metastasis, and patients with different mutation spectra are considered as multiple primary cancers.

[0024] The second group of research sample information is as follows:

[0025] Including 52 clinical sample data, the data is derived from the actual samples obtained by the patent applicant in clinical research. The 52 samples are samples of radical esophagectomy for esophageal tumors at the Chinese Academy of Medical Sciences from September 2022 to August 2024. The postoperative tumor tissue was sent to the pathology department for diagnosis. All patients had two independent esophageal squamous cell carcinoma lesions, and according to the existing tissue pathology diagnosis standards, they can be clearly diagnosed as multiple primary squamous cell carcinoma and intramural metastatic squamous cell carcinoma. The diagnosis was made by two different experienced pathologists, and the diagnosis of the two must be consistent. This sample can be used to verify the classification effect of the technical solution in this patent. There are a total of 52 multifocal squamous cell carcinoma samples in the verification sample, including 23 multiple primary squamous cell carcinomas and 29 esophageal squamous cell carcinomas with intramural metastasis. The verification sample has also been sequenced in different cancer foci of tumor tissue samples as in this patent, and mutation information is obtained and analyzed based on a panel composed of 20 genes.

[0026]

[0027]

[0028]

[0029]

[0030] Among them, 5 samples had no mutation detection, and it was impossible to determine whether they were metastatic or primary. Taking the tissue diagnosis results as the gold standard, based on the original data of this patent, it can be calculated that the diagnostic sensitivity of the technical solution in this patent for multiple primary cancers is 87.0%, and the specificity is 100%; the diagnostic sensitivity of intramural metastasis is 93.1%, and the specificity is 100%. The consistency of the molecular detection results of this technical solution compared with the gold standard is 47 / 52=90.4%. It can be seen that if it is determined to be intramural metastasis, the mutation types between the two are completely consistent, but if it is multiple primary squamous cell carcinoma, the two are different. According to the principle of tumor clonal evolution, multiple primary cancers originate from different tumor stem cells, so there are different gene variation spectra between different cancer foci; while the primary foci and metastatic foci of metastatic cancer originate from the same tumor stem cell, so the gene variation spectra between different cancer foci will be the same or similar.

Claims

1. Use of a mutation detection reagent of a gene marker in the preparation of a diagnostic reagent for distinguishing primary and intramural metastasis of multifocal squamous cell carcinoma of the esophagus, characterized in that: The gene markers are composed of the following 20 genes: TP53, TTN, NOTCH1, PCLO, DAMTSL3, ZNF750, DNAH3, EP300, CDKN2A, MBTPS1, MYH15, RHBG, RLF, SYNE1, XIRP2, C6orf118, EYS, FILIP1L, LTBP1, and NCOA2.

2. The use according to claim 1, characterized in that: The mutation detection reagent is used to detect exon mutations of gene markers.

3. A detection device, characterized in that: It is used to differentiate primary and intramural metastases of multifocal squamous cell carcinoma of the esophagus, including: Gene detection module, used to detect mutations in the following 20 genes: TP53, TTN, NOTCH1, PCLO, DAMTSL3, ZNF750, DNAH3, EP300, CDKN2A, MBTPS1, MYH15, RHBG, RLF, SYNE1, XIRP2, C6orf118, EYS, FILIP1L, LTBP1, NCOA2; The comparison module compares the information obtained by the gene detection module on different esophageal cancer foci of the same person to be tested. If the types of gene mutations in different cancer foci are the same, it is intramural metastasis; if they are different, it is multiple primary cancers.

4. A computer-readable medium, characterized in that: It is used to distinguish primary and intramural metastasis of multifocal squamous cell carcinoma of the esophagus, and it records a computer program that can run the following method: Step 1, obtaining mutation information of gene markers on different cancer lesions of the same person to be tested, wherein the gene markers are composed of the following 20 genes: TP53, TTN, NOTCH1, PCLO, DAMTSL3, ZNF750, DNAH3, EP300, CDKN2A, MBTPS1, MYH15, RHBG, RLF, SYNE1, XIRP2, C6orf118, EYS, FILIP1L, LTBP1, NCOA2; Step 2: Obtain the gene mutation spectra of different cancer foci. If they are the same, it is judged as intramural metastasis; if they are different, it is multiple primary cancers.

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