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Application of gene FOXD4 to preparation of diagnosis reagent kit for acute myelocytic leukemia and reagent kit

A diagnostic kit and acute myeloid technology, applied in the fields of biomedical technology and hematological leukemia, can solve the problem of no research report on the expression of FOXD4 gene family members

Pending Publication Date: 2019-12-20
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no research on the expression of FOXD4 gene family members in bone marrow and peripheral blood nucleated cells of clinical patients with acute myeloid leukemia (AML).

Method used

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  • Application of gene FOXD4 to preparation of diagnosis reagent kit for acute myelocytic leukemia and reagent kit
  • Application of gene FOXD4 to preparation of diagnosis reagent kit for acute myelocytic leukemia and reagent kit
  • Application of gene FOXD4 to preparation of diagnosis reagent kit for acute myelocytic leukemia and reagent kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] We first validated Kras in an AML animal model G12D / + ; In CD2Cre transgenic mice, myeloid / monocytic progenitor cells in the bone marrow and myeloid leukemia cells in the spleen up-regulated the FOXD4 signaling pathway gene ( figure 1 ). (A, B) Quantitative qPCR to determine the difference in transcript levels of candidate genes. RNA from control and Kras G12D / + ; CD2Cre + myeloid / monocytic progenitors in bone marrow (A) or Gr1 in spleen + Mac1 + Extracted from myeloid leukemia cells (B). The relative expression level is expressed as the expression fold relative to GAPDH.

[0044] The result was that qPCR verified the gene expression of the granulocyte TGF-β pathway in AML mice was elevated ( figure 1 A, B). qPCR results proved that Tgfβr1, Tgfβ1, Foxd4 in Kras G12D / + ; CD2Cre + myeloid / monocytic progenitors and Gr1 + Mac1 + Highly expressed in myeloid leukemia cells ( figure 1 A, B), while the expression of Hoxa3, foxa2, and foxa3 are less different.

Embodiment 2

[0046] To evaluate the gene expression level of FOXD4 in peripheral blood of clinical acute myeloid leukemia patients.

[0047] Experimental materials: Collect peripheral blood samples from 8 patients with acute myeloid leukemia, and collect peripheral blood samples from 3 normal people at the same time, and use fluorescence real-time quantitative PCR to carry out classical molecular biology experiment verification (qRT-PCR).

[0048] Experimental results: The target band was determined by melting curve analysis and electrophoresis, and the ΔΔCT method was used for relative quantification. The results are as follows: figure 2 shown. Compared with normal peripheral blood samples, the expression of FOXD4 gene family members FOXD4(A) and FOXD4L1(B) in the peripheral blood of 3 clinical myeloid leukemia patients was up-regulated by q-PCR. There was no significant difference between FOXD4L4 and FOXD4L5.

Embodiment 3

[0050] The expression of FOXD4 in myeloid leukemia patients in Oncomine database.

[0051] We found in the Oncomine database that the expression of FOXD4 gene was positively correlated with the exacerbation stage of acute myeloid leukemia and chronic myeloid leukemia ( image 3 ).

[0052]

[0053]

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Abstract

The invention relates to an application of a gene FOXD4 to preparation of a diagnosis reagent kit for acute myelocytic leukemia and a reagent kit. The diagnosis reagent kit comprises an SYBR Green polymerase chain reaction system, and a primer pair for amplifying the FOXD4 gene and a house-keeping gene, wherein the SYBR Green polymerase chain reaction system comprises a PCR buffer solution, dNTPsand an SYBR Green fluorescent dye. The FOXD4 as a detection and diagnosis target point is applied to the diagnosis reagent kit which is clinically prepared, so that peripheral blood or bone marrow karyocyte of a patient suffering from the acute myeloid leukemia can be quickly diagnosed or forecasted; and the transcriptional level of the target gene FOXD4 in samples is detected, so that whether anexaminee suffers from the acute myeloid leukemia can be judged, and a basis is provided for early diagnosis and treatment of the cancer.

Description

technical field [0001] The invention belongs to the fields of biomedical technology and hematological leukemia, and relates to a nucleic acid detection kit and its application in in vitro diagnosis of acute myeloid leukemia. Background technique [0002] Acute myeloid leukemia (AML) is one of the diseases with high mortality caused by malignant changes of hematopoietic stem cells, and has a high degree of malignancy in cancer cases all over the world. During the course of treatment, radiation therapy, chemical drug therapy, bone marrow transplantation, etc. are generally used, but there are certain problems in terms of clinical success rate, transplant matching and prognosis. Early diagnosis of leukemia is very important in the treatment of the disease, so finding and identifying effective biomarkers and even serving as therapeutic targets is an urgent problem to be solved. Molecular diagnostic technology has a unique objective advantage in judging AML, and the in vitro det...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/158C12Q2600/166
Inventor 刘菲菲刘伟郑文杰王松刘凤勇李莹慧檀露王岩
Owner NANKAI UNIV