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CN-122012717-A - Molecular marker combination for diagnosing uterine leiomyosarcoma, application thereof, kit and application thereof

CN122012717ACN 122012717 ACN122012717 ACN 122012717ACN-122012717-A

Abstract

The invention discloses a molecular marker combination for diagnosing uterine leiomyosarcoma and application thereof, a kit and application thereof, wherein the marker combination is a combination of (i), (ii) and (iii), wherein (i) is a CKS2 gene or a protein encoded by the CKS2 gene, (ii) is a TOP2A gene or a protein encoded by the TOP2A gene, and (iii) is a TRPC4 gene or a protein encoded by the TRPC4 gene. The invention also discloses application of the molecular marker combination in preparing a detection kit for diagnosis, auxiliary diagnosis or prognosis evaluation of uterine leiomyosarcoma. The invention discovers that the three genes have specific expression change in ULMS tissues through multi-group transcriptome data analysis and immune group verification, and can be used as a reliable index for distinguishing ULMS from benign lesions. The marker combination has the advantages of high diagnosis accuracy, wide application and remarkable clinical transformation value.

Inventors

  • LI JING
  • YU ZHIYING
  • XIAO ZHIJIE

Assignees

  • 浙江大学医学院附属妇产科医院(浙江省妇女医院、浙江省妇女保健院)

Dates

Publication Date
20260512
Application Date
20260318

Claims (10)

  1. 1. A combination of molecular markers for diagnosing uterine leiomyosarcoma, characterized in that the combination of molecular markers is a combination of (i), (ii) and (iii), wherein: (i) Is the CKS2 gene or a protein encoded by the same; (ii) A TOP2A gene or a protein encoded by the TOP2A gene; (iii) Is a TRPC4 gene or a protein encoded by the same.
  2. 2. The molecular marker combination for diagnosing uterine leiomyosarcoma according to claim 1, wherein Gene ID of the CKS2 Gene in NCBI is 1164, gene ID of the TOP2A Gene in NCBI is 7153, and Gene ID of the TRPC4 Gene in NCBI is 7223.
  3. 3. Use of a combination of molecular markers for the diagnosis of uterine leiomyosarcoma, characterized in that the combination of molecular markers for the diagnosis of uterine leiomyosarcoma according to claim 1 is used for diagnosis, auxiliary diagnosis or prognostic evaluation of uterine leiomyosarcoma.
  4. 4. A kit, characterized in that the kit comprises: a) Reagents for detecting the CKS2 gene or a protein encoded thereby; b) Reagents for detecting the TOP2A gene or a protein encoded thereby; c) An agent for detecting TRPC4 gene or a protein encoded thereby; d) Normal control samples.
  5. 5. The kit of claim 4, wherein the reagent for detecting the CKS2 gene is a specific nucleic acid probe or primer targeting mRNA transcribed from the CKS2 gene, the reagent for detecting the TOP2A gene is a specific nucleic acid probe or primer targeting mRNA transcribed from the TOP2A gene, the reagent for detecting the TRPC4 gene is a specific nucleic acid probe or primer targeting mRNA transcribed from the TRPC4 gene; The reagents for detecting the protein encoded by the CKS2 gene, the protein encoded by the TOP2A gene and the protein encoded by the TRPC4 gene are antibodies corresponding to the protein encoded by the CKS2 gene, the protein encoded by the TOP2A gene and the protein encoded by the TRPC4 gene, respectively.
  6. 6. The kit of claim 5, wherein the sequences of primers targeting mRNA transcribed from the CKS2 gene are shown in SEQ ID NO.1 and SEQ ID NO.2, respectively, the sequences of primers targeting mRNA transcribed from the TOP2A gene are shown in SEQ ID NO.3 and SEQ ID NO.4, respectively, and the sequences of primers targeting mRNA transcribed from the TRPC4 gene are shown in SEQ ID NO.5 and SEQ ID NO.6, respectively.
  7. 7. The kit of claim 6, further comprising reagents for RNA extraction, reverse transcription and qPCR detection.
  8. 8. Use of a kit according to claim 4 for distinguishing whether a sample to be tested originates from a subject suffering from uterine leiomyosarcoma, the method of use of the kit comprising the steps of: 1) Collecting a tissue sample or a body fluid sample of a subject to obtain a sample to be tested; 2) Detecting the expression amounts of CKS2 gene, TOP2A gene and TRPC4 gene in the sample to be detected by using the kit according to claim 4 and adopting a molecular biological means, wherein the expression amounts are the content of mRNA transcribed from the gene or the content of protein coded by the gene; 3) Comparing the detection result with the expression amounts of the CKS2 gene, the TOP2A gene and the TRPC4 gene in a normal control sample; 4) And distinguishing whether the sample to be tested is derived from a subject suffering from uterine leiomyosarcoma according to the difference of the gene expression quantity.
  9. 9. The use of the kit according to claim 8, wherein in step 2), the molecular biological means for detecting the expression levels of CKS2 gene, TOP2A gene and TRPC4 gene in the sample to be tested comprises real-time fluorescent quantitative PCR, digital PCR, RNA in situ hybridization, immunohistochemical detection and enzyme linked immunosorbent assay.
  10. 10. The use of the kit according to claim 9, wherein in step 3), the test sample is derived from a subject suffering from uterine leiomyosarcoma when both the expression level of CKS2 gene and the expression level of TOP2A gene in the test sample are increased compared to those in a normal control sample, and the expression level of TRPC4 gene in the test sample is decreased compared to those in a normal control sample.

Description

Molecular marker combination for diagnosing uterine leiomyosarcoma, application thereof, kit and application thereof Technical Field The invention relates to the technical field of medicines. In particular to a molecular marker combination for diagnosing uterine leiomyosarcoma and application thereof, a kit and application thereof. Background Uterine leiomyosarcoma (Uterine Leiomyosarcoma, ULMS) is a rare but highly malignant tumor of myometrial origin, accounting for about 2% -5% of uterine malignancies, with an incidence of about 1 out of hundred thousand people per year. ULMS has extremely strong invasiveness and metastatic capacity, shows high mortality, high recurrence rate and broad drug resistance, and has extremely poor prognosis. Its clinical symptoms are very similar to common benign uterine leiomyomas (Uterine Leiomyoma, ULM), such as irregular vaginal bleeding, anemia, and abdominal mass, etc., resulting in difficulty in accurate diagnosis before ULMs surgery. At present, the clinical diagnosis of ULMS mainly depends on the pathological examination of the postoperative frozen section and the postoperative paraffin, lacks sensitive and specific preoperative diagnosis markers, and has unsatisfactory sensitivity and specificity of common indexes such as CA125, MRI, ki-67 and the like. ULMS treatment approaches are still mainly surgical excision, auxiliary chemotherapy or radiotherapy schemes have limited survival improvement, and targeted treatment and immunotherapy have not been progressed. However, misdiagnosis of uterine leiomyosarcoma as benign uterine fibroid prior to surgery may lead to the spread of sarcoma cells during resection using an electric rotary cutter, severely affecting patient prognosis. In view of the above, there is a need to develop a molecular marker combination that is able to sensitively and specifically distinguish ULMS from ULM to improve preoperative diagnostic accuracy and aid in predicting prognosis to optimize therapeutic decisions and follow-up protocols. Disclosure of Invention Therefore, the technical problem to be solved by the invention is to provide a molecular marker combination for diagnosing uterine leiomyosarcoma, application thereof, a kit and application thereof, wherein the related molecular marker combination is developed based on transcriptome integrated analysis, shows higher diagnosis accuracy and good prognosis prediction value in a plurality of groups of data sets, prompts the molecular marker combination to be used as a clinical application potential of the molecular marker group for diagnosing uterine leiomyosarcoma, can be used for early auxiliary diagnosis of ULMS, can assist in carrying out layered evaluation on prognosis of patients, and provides support for personalized treatment. In order to solve the technical problems, the invention provides the following technical scheme: a combination of molecular markers for diagnosing uterine leiomyosarcoma, said combination of molecular markers being a combination of (i), (ii) and (iii), wherein: (i) Is the CKS2 gene or a protein encoded by the same; (ii) A TOP2A gene or a protein encoded by the TOP2A gene; (iii) Is a TRPC4 gene or a protein encoded by the same. The CKS2 gene, TOP2A gene and TRPC4 gene in the molecular markers specifically refer to the mRNA produced by their transcription, which may include transcripts of different splice forms. The different transcripts can all be used as components in a combination of molecular markers for diagnosing uterine leiomyosarcoma. Furthermore, the proteins produced by translation of the gene and natural variants or different translation subtypes thereof can also be used as components in a combination of molecular markers for diagnosing leiomyosarcoma in the uterus. Although single or double marker detection systems exist on the market, the combined detection of three molecular markers can effectively eliminate false positives caused by heterogeneous expression (only up-regulated CKS2, only up-regulated TOP2A or only down-regulated TRPC 4) of other tumors, and improve the accuracy of ULMS differential diagnosis. The above molecular marker combination for diagnosing uterine leiomyosarcoma, gene ID 1164 of the CKS2 Gene in NCBI, gene ID 7153 of the TOP2A Gene in NCBI, and Gene ID 7223 of the TRPC4 Gene in NCBI. Use of a combination of molecular markers for diagnosing uterine leiomyosarcoma, the combination of molecular markers for diagnosing uterine leiomyosarcoma described above being used for diagnosis, auxiliary diagnosis or prognostic evaluation of uterine leiomyosarcoma. A kit, the kit comprising: a) Reagents for detecting the CKS2 gene or a protein encoded thereby; b) Reagents for detecting the TOP2A gene or a protein encoded thereby; c) An agent for detecting TRPC4 gene or a protein encoded thereby; d) Normal control samples. The reagent used for detecting the CKS2 gene is a specific nucleic acid probe or primer targeting mRNA transcribed from the CKS2