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Method for enriching tumor stem cells through continuous low dose radiation

A tumor stem cell, low-dose technology, applied in the biological field, can solve the problems of difficult sample acquisition and small proportion of tumor stem cells.

Inactive Publication Date: 2014-11-19
BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The proportion of tumor stem cells in tumors is extremely small. Using existing isolation methods, such as using specific antigen molecular markers on the surface of specific tumor cells, only a very small amount of tumor stem cells can be obtained from a large number of tumor tissues or tumor cell lines for biological Scientific research, in some experiments that require a large amount of samples, it is more difficult to obtain samples

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  • Method for enriching tumor stem cells through continuous low dose radiation
  • Method for enriching tumor stem cells through continuous low dose radiation
  • Method for enriching tumor stem cells through continuous low dose radiation

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Example 1 Enrichment of MDA-MB-231 tumor stem cells

[0029] The breast cancer cell line MDA-MB-231 was cultured under routine laboratory conditions, after isotope 60 Co radiation source, 2Gy irradiation treatment (1:17min at a dose of 139.09cGy / min), and routine culture for 1-2 weeks after treatment, when the cell growth condition is stable, accept again 60 Co and 2Gy irradiation treatments were repeated, and cells (n*2Gy) irradiated with 1*2Gy, 2*2Gy and 3*2Gy were collected respectively.

Embodiment 2

[0030] Example 2 Enrichment of MCF7 tumor stem cells

[0031] The breast cancer cell line MCF7 was cultured under routine laboratory conditions, after isotope 60 Co radiation source, 2Gy irradiation treatment (1:17min at a dose of 139.09cGy / min), and routine culture for 1-2 weeks after treatment, when the cell growth condition is stable, accept again 60 Co and 2Gy irradiation treatments were repeated, and cells (n*2Gy) irradiated with 1*2Gy, 2*2Gy and 3*2Gy were collected respectively.

Embodiment 3

[0032] Example 3 Flow cytometry analysis of enrichment of tumor stem cells

[0033] The cells obtained in Examples 1 and 2 were incubated with two antibodies, CD24-APC and CD44-FITC, respectively, and flow cytometry analysis found that continuous low-dose radiation in the two cell lines could transform CD44 + CD24 - / low Especially in the MCF7 cell line, the proportion of tumor stem cells increased by nearly 10 times, thus indicating that the use of continuous low-dose radiation breast cancer cell lines can increase the proportion of tumor stem cells (see figure 1 ).

[0034] The above-mentioned flow cytometry experiment process is briefly described as follows: using the method of flow cytometry sorting, firstly collect different batches of processed cells, wash them with phosphate buffered solution PBS, and then use two kinds of CD24-APC and CD44-FITC The cells were incubated with the antibody, and 635nm and 488nm excitation light were selected respectively, and the quadrant...

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Abstract

The invention relates to a method for enriching tumor stem cells through continuous low dose radiation. The method comprises the following steps: culturing cancer cell lines, carrying out low dose radiation, carrying out the low dose radiation again after culturing for a period of time, radiating for N times in total to enrich the tumor stem cells in the cell lines and increase the ratio of the tumor stem cells, wherein the cancer cell lines are breast cancer cell lines preferably; 60 Co is preferably adopted to carry out the low dose radiation; the low dose is 1.5-3Gy preferably and 2Gy more preferably; the culturing time is 1-2 weeks preferably; and the radiating times in total is 3 preferably. The method for enriching the tumor stem cells, provided by the invention, is very simple and practical and a new way is developed for the follow-up further study of the characteristics of the tumor stem cells.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for continuously enriching tumor stem cells with low-dose radiation. Background technique [0002] Tumor is a collection of heterogeneous cells, among which there is a subgroup of cells with strong self-renewal ability and tumorigenic ability, which are defined as cancer stem cells (Cancer Stem Cells). The theory of cancer stem cells believes that there is a certain amount of tumor stem cells in the primary tumor lesion. After conventional treatments such as surgery, chemotherapy and radiotherapy, although almost all tumor cells can be killed, there are still some tumor stem cells remaining. Stem cells have a strong self-renewal ability, and will regenerate tumors at the original lesion, causing disease recurrence. At the same time, tumor stem cells can transfer in the body and form metastases (see Vermeulen, L., Sprick, M.R., Kemper, K., Stassi , G. & Medema, J.P. Cancer s...

Claims

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Application Information

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IPC IPC(8): C12N13/00C12N5/095
Inventor 孙英丽于慧
Owner BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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