Authenticating method of cancer stem cell

A cancer stem cell and identification method technology, applied in the identification field of cancer stem cells, can solve the problems of leukemia recurrence, difficult bone marrow source, etc.

Inactive Publication Date: 2015-09-23
冯宝章
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Problems solved by technology

[0003] Bone marrow transplantation is an effective way to treat acute leukemia, but the source of bone marrow is very difficult
There is no difficulty in the source of auto

Method used

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  • Authenticating method of cancer stem cell
  • Authenticating method of cancer stem cell
  • Authenticating method of cancer stem cell

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Embodiment Construction

[0022] The above and other technical features and advantages of the present invention will be described in more detail below in conjunction with the accompanying drawings.

[0023] Based on the above research, the identification method of cancer stem cells of the present invention is:

[0024] In situ hybridization (ISH) was performed on bone marrow smears with two oncogene oligonucleotides labeled with biotin. The specific process is:

[0025] Step 1: Anticoagulate 4ml of the bone marrow aspiration specimen in a heparin tube, collect the lymphocyte suspension in an EP tube by centrifugation, and then make a smear on a smear machine;

[0026] Step 2, V-erbB Oligo probe labeling: pre-treatment of the cell smear before hybridization, pre-hybridization and hybridization, washing and detection after hybridization, etc. are all carried out according to the requirements of the kit.

[0027] 1) Observation of results: 200 nucleated cells were counted under a microscope. Brown spot...

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Abstract

The invention relates to an authenticating method of a cancer stem cell. Oligonucleotides of pathogenic cancer gene and cancerous cancer gene are marked by biotins to be subjected to in-situ hybridization, and the cell with dual-positive result is identified to be the cancer stem cell. The authenticating method of the cancer stem cell comprises the following step of performing in-situ hybridization on a to-be-detected bone marrow cell smear, wherein the used probe is V-erbB (virus oncogene) and TS gene oligonucleotides marked by digoxin, and the V-erbB oligonucleotide is the oligonucleotide generating mutation/amplification location. The authenticating method can be used as a basis for judging whether the bone marrow of an acute leukemia complete remission (AL-CR) patient is normal or not, so that collection of normal bone marrow is determined and is stored at a low temperature for later use. When leukemia relapse occurs on the AL-CR patient, the normal bone marrow can be used for effective treatment. The other technology in the study, i.e. a V-erbB and TS gene PCR (polymerase chain reaction) gene diagnosis method is combined to detect a specimen, and a negative reaction result supports judgment that whether the bone marrow is normal or not.

Description

technical field [0001] The invention relates to the field of cancer diagnosis, in particular to a method for identifying cancer stem cells. Background technique [0002] At present, the understanding of stem cells stays in their function, rather than in the shape of the naked eye, which brings many difficulties. In particular, the identification and amount of leukemia stem cells in the bone marrow are particularly important for autologous bone marrow transplantation in the treatment of leukemia relapse in patients with AL-CR. Doctors should judge whether the patient’s bone marrow is normal at the AL-CR stage, that is, whether it contains or not or how many leukemia stem cells it contains, and decide whether to collect and store it at low temperature, so as to evaluate whether the bone marrow can be used for autologous bone marrow transplantation in the future to treat leukemia relapse. [0003] Bone marrow transplantation is an effective way to treat acute leukemia, but the...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 冯宝章
Owner 冯宝章
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