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CN-122012708-A - Aflatoxin-related liver cancer marker and application thereof

CN122012708ACN 122012708 ACN122012708 ACN 122012708ACN-122012708-A

Abstract

The invention provides an aflatoxin-related liver cancer marker and application thereof, belonging to the technical field of molecular diagnosis. The invention obtains a diagnosis and prognosis judgment marker of aflatoxin-related liver cancer parting, namely FBXW7 gene chr 4:152326101C > T by research methods and means such as a large-sample high-throughput gene sequencing technology, a TaqMan-PCR technology, a cytology technology, experimental zoology and the like. Experiments show that in the aflatoxin-related liver cancer cells, aflatoxin is closely related to biological behaviors such as cancer cell proliferation, migration, invasion, cell cycle, drug resistance and the like, and the relationship is influenced by the FBXW7 gene chr 4:152326101C > T. The marker and the establishment of a related evaluation technology system provide important data support for refining pathological characteristics of liver cancer molecules.

Inventors

  • LONG XIDAI
  • ZHU XIAOYING
  • LONG QINQIN
  • LIANG QIUJU
  • Wang Xingzhizi

Assignees

  • 武汉双旋生物科技有限责任公司

Dates

Publication Date
20260512
Application Date
20260122

Claims (10)

  1. 1. A marker for liver cancer related to aflatoxin is characterized in that GRCh38 is taken as a reference genome, and the marker is FBXW7 gene chr4:152326101C > T.
  2. 2. Use of the marker of claim 1 or an expressed product thereof for preparing a detection product or/and a prognosis evaluation product for aflatoxin-related liver cancer typing.
  3. 3. The use according to claim 2, wherein the detection product is a kit.
  4. 4. A TaqMan-PCR kit for detecting the aflatoxin-related liver cancer marker according to claim 1 is characterized in that the TaqMan-PCR kit comprises a primer pair for amplifying an FBXW7 gene, a probe T shown as SEQ ID No.17 and a probe C shown as SEQ ID No.18, wherein the sequences of the primer pair are shown as SEQ ID No.19 and SEQ ID No. 20.
  5. 5. The TaqMan-PCR kit according to claim 4, wherein the TaqMan-PCR kit comprises a positive reference substance and a standard substance, the nucleotide sequence of the positive reference substance is shown as SEQ ID No.21 and SEQ ID No.22, and the nucleotide sequence of the standard substance is shown as SEQ ID No.23 and SEQ ID No. 24.
  6. 6. The TaqMan-PCR kit according to claim 4, wherein the TaqMan-PCR kit comprises a PCR amplification reagent consisting of 0.08U/. Mu.L Taq deoxyribonucleic acid polymerase, 2U/. Mu.L enzyme buffer, 0.4. Mu.M deoxyribonucleotide triphosphate and 4. Mu.M magnesium chloride.
  7. 7. A PCR kit for amplifying the aflatoxin-related liver cancer marker according to claim 1 is characterized by comprising an amplicon primer pair, wherein the gene fragment of the amplicon comprises the marker according to claim 1, and the sequences of the amplicon primer pair are shown as SEQ ID No.25 and SEQ ID No. 26.
  8. 8. The PCR kit of claim 7, wherein the PCR kit comprises a positive control having nucleotide sequences as set forth in SEQ ID No.27 and SEQ ID No. 28.
  9. 9. The PCR kit of claim 7, wherein the PCR kit comprises an amplicon PCR amplification reagent comprising 0.08U/. Mu.L Taq deoxyribonucleic acid polymerase, 2U/. Mu.L enzyme buffer, 0.4. Mu.M deoxyribonucleotide triphosphate, and 4. Mu.M magnesium chloride.
  10. 10. The PCR kit of claim 7, wherein the PCR kit comprises an amplicon purification reagent comprising 3M sodium acetate at pH 5.2, absolute ethanol, and TE buffer at a concentration of 10mM Tris-HCl and 1mM ethylenediamine diacetic acid sodium salt.

Description

Aflatoxin-related liver cancer marker and application thereof Technical Field The invention belongs to the technical field of molecular diagnosis, and particularly relates to an aflatoxin-related liver cancer marker and application thereof. Background Aflatoxin is a class I chemical carcinogen capable of inducing liver cancer, and the liver cancer induced by aflatoxin (namely, aflatoxin-related liver cancer) has high incidence rate in southeast coastal areas of China such as Guangxi and Guangdong areas, and seriously affects the physical health of residents. Epidemiological and toxicological studies have demonstrated that aflatoxins are metabolites of aspergillus flavus and aspergillus parasiticus, which are relatively easy to grow and reproduce in a humid and hot environment, and therefore, a large amount of these microorganisms and their metabolites aflatoxins exist in foods such as peanuts, corns, etc. which are poorly preserved in a humid and hot environment. Aflatoxin enters the human body along with the grains polluted by the aflatoxin, and is mainly metabolized in the liver. After entering the nucleus of liver cells, aflatoxin often forms aflatoxin-DNA adducts, which can cause DNA damage, cause unstable genome structure, and further induce malignant transformation of liver cells. Research shows that in the development and progress of aflatoxin-related liver cancer, specific molecular changes exist. For example, in tumor tissue, there is a relatively high frequency of point mutations at codon 249 of the TP53 gene, which are positively correlated with aflatoxin exposure levels. With the advancement of high throughput sequencing technology and genome planning, some significant genetic heterogeneity features were found in aflatoxin-related liver cancers. However, in terms of research on application value of aflatoxin-related liver cancer molecular typing markers and clinical treatment and prognosis judgment, no report is currently available. Disclosure of Invention In order to solve the problems, the first aspect of the invention provides an aflatoxin-related liver cancer marker, GRCh38 is taken as a reference genome, and the marker is FBXW7 gene chr4:152326101C > T. The second aspect of the invention provides the application of the aflatoxin-related liver cancer marker or the expression product thereof in preparing a detection product and/or a prognosis evaluation product for aflatoxin-related liver cancer typing. In a preferred embodiment, the detection product is a kit. The third aspect of the invention provides a TaqMan-PCR kit for detecting the aflatoxin-related liver cancer marker, wherein the TaqMan-PCR kit comprises a primer pair for amplifying an FBXW7 gene, a probe T shown as SEQ ID No.17 and a probe C shown as SEQ ID No.18, and the sequences of the primer pair are shown as SEQ ID No.19 and SEQ ID No. 20. In a preferred scheme, the TaqMan-PCR kit comprises a positive control and a standard substance, wherein the nucleotide sequence of the positive control is shown as SEQ ID No.21 and SEQ ID No.22, and the nucleotide sequence of the standard substance is shown as SEQ ID No.23 and SEQ ID No. 24. In a preferred embodiment, the TaqMan-PCR kit comprises PCR amplification reagents consisting of 0.08U/. Mu.L Taq deoxyribonucleic acid polymerase, 2U/. Mu.L enzyme buffer, 0.4. Mu.M deoxyribonucleotide triphosphate and 4. Mu.M magnesium chloride. In a preferred scheme, the DNA sample to be detected of the TaqMan-PCR kit is a liver cancer tumor tissue source, and the specific extraction method comprises the steps of taking a proper amount of tumor tissue, separating cells by using proteinase K, and extracting DNA in the cells by using a phenol-chloroform extraction method as the DNA to be detected. The fourth aspect of the invention provides a PCR kit for amplifying the aflatoxin-related liver cancer marker, which comprises an amplicon primer pair, wherein the gene fragment of the amplicon comprises the marker of claim 1, and the sequences of the amplicon primer pair are shown as SEQ ID No.25 and SEQ ID No. 26. In a preferred embodiment, the PCR kit comprises a positive control, wherein the nucleotide sequence of the positive control is shown as SEQ ID No.27 and SEQ ID No. 28. In a preferred embodiment, the PCR kit comprises an amplicon PCR amplification reagent comprising the following components of 0.08U/. Mu.L Taq deoxyribonucleic acid polymerase, 2U/. Mu.L enzyme buffer, 0.4. Mu.M deoxyribonucleotide triphosphate, and 4. Mu.M magnesium chloride. In a preferred embodiment, the PCR kit comprises an amplicon purification reagent comprising 3M sodium acetate at pH 5.2, absolute ethanol and TE buffer at a concentration of 10mM Tris-HCl and 1mM ethylenediamine diacetate. In a preferred scheme, the DNA sample to be detected of the PCR kit is a liver cancer tumor tissue source, and the specific extraction method comprises the steps of taking a proper amount of tumor tissue, separating cells by using proteinas