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Screening method and application of tumor metastasis starting cells

A technology for initiating cells and tumor metastasis, applied in the field of cell screening, it can solve the problems of few tumor types, low enrichment rate, long time-consuming, etc., and achieve the effects of simple operation, high enrichment efficiency, and wide application.

Pending Publication Date: 2022-07-29
THE HONG KONG POLYTECHNIC UNIV SHENZHEN RES INST
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a screening method and application of tumor metastasis-initiating cells in view of the deficiencies in the prior art, aiming to solve the problems of long time-consuming and low enrichment rate in the existing methods for screening metastasis-initiating cells And the problem of fewer tumor types that can be screened

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  • Screening method and application of tumor metastasis starting cells
  • Screening method and application of tumor metastasis starting cells
  • Screening method and application of tumor metastasis starting cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Breast cancer cells SKBR3 cells were suspended in cell culture medium and loaded into the circulatory system for 12h at 20dyne / cm 2 After blood flow shear stress treatment, its transfer ability and self-renewal ability were tested.

[0049] The detection step of metastasis ability is as follows: add the same number of cells in the control group and the treatment group to the upper layer of the 8 μm pore size Transwell chamber containing the culture medium containing low serum. Due to the attraction of the high serum in the lower layer, the tumor cells with strong migration ability can pass through the chamber to reach the transwell chamber. lower end.

[0050] The result is as image 3 shown, tumor cells treated with blood flow shear stress versus untreated cells (Control) and cells treated only in suspension (0 dyne / cm 2 ) more cells reached the bottom of the chamber, indicating a stronger migration ability.

[0051] The steps of cell self-renewal ability detection ...

Embodiment 2

[0054] The same number of tumor cells (20 dyne / cm 2 ), only suspension-treated cells (0 dyne / cm 2 ), untreated control cells (Control), and positive control cells (Tic) cultured by the tumor stem cell culture method were injected subcutaneously in nude mice, and tumor growth was measured regularly after 14-34 days of growth. .

[0055] The result is as Figure 5 shown, 20dyne / cm 2 Tumor cells treated with blood flow shear stress have stronger tumor-forming ability in nude mice.

Embodiment 3

[0057] The CXCR4 pathway plays an important role in the process of tumor metastasis. In this example, different concentrations of CXCR4 inhibitor-AMD3465 were selected to inhibit the activity of the CXCR4 pathway. Tumor cells were first treated with blood flow shear stress for 12 hours, then added with different concentrations of inhibitors and cultured for 24 hours, and then their metastatic ability was detected by Transwell chamber.

[0058] The result is as Image 6 As shown, with the increase of inhibitor concentration, the migration ability of the selected tumor-initiating cells also decreased gradually. It was shown that inhibiting the CXCR4 pathway can significantly inhibit the metastasis ability of the screened metastasis-initiating cells.

[0059] To sum up, the present invention mainly uses a peristaltic pump to generate blood flow shear stress that meets physiological conditions to screen metastasis-initiating cells that can effectively generate metastatic tumors. ...

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Abstract

The invention discloses a screening method and application of tumor metastasis starting cells, and the screening method comprises the following steps: carrying out shearing treatment on tumor cells by adopting blood flow shearing forces with different intensities, and collecting and recording the treated tumor cells in different time periods; performing function detection related to metastasis starting cells on the treated tumor cells, and determining an optimal screening condition according to a detection result; and processing the tumor cells to be screened based on the optimal screening condition to obtain the tumor metastasis starting cells. The method provided by the invention has the advantages of simplicity in operation, wide application, high enrichment efficiency and the like. The method can be applied to metastasis starting cell screening of different tumor stages and different tumor types; according to the method, a part of malignant and weak metastasis starting cells which cannot survive under the blood flow shear stress can be killed, and meanwhile, a part of cells with weak metastasis capacity are converted into tumor cells with strong metastasis capacity, so that the enrichment efficiency is high.

Description

technical field [0001] The invention relates to the technical field of cell screening, in particular to a screening method and application of tumor metastasis initiating cells. Background technique [0002] Metastasis-initiating cells are a rare subset of tumor cells that can significantly form distant metastases. Therefore, it is of great significance to effectively isolate metastasis-initiating cells. Currently, the isolation of metastasis-initiating cells is mainly based on two methods: first, based on cell surface markers, which are limited by specific artificially selected markers and are difficult to be universally applied in different tumor types; second, injection of tumor cells In animals, an in vivo screening method of extracting cells after distant metastasis has been developed, which requires a longer time and has a lower enrichment rate. [0003] Therefore, the existing technology still needs to be improved and developed. SUMMARY OF THE INVENTION [0004] Th...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02
CPCC12N5/0693C12N5/0631G01N33/5005C12N2509/10
Inventor 谭又华辛影
Owner THE HONG KONG POLYTECHNIC UNIV SHENZHEN RES INST