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Screening kit of malignant breast cancer stem cells

A technology of breast cancer stem cells and kits, applied in the field of malignant breast cancer stem cell screening kits, has achieved good application prospects

Inactive Publication Date: 2013-12-11
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Screening kit of malignant breast cancer stem cells
  • Screening kit of malignant breast cancer stem cells
  • Screening kit of malignant breast cancer stem cells

Examples

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Effect test

Embodiment 1

[0029] Example 1 Study on the relationship between CXCR6 gene expression and malignant phenotype of breast cancer stem cells

[0030] 1. Experimental method

[0031] (1) Cell sorting and detection of CXCR6 gene expression in various cells

[0032] Take the human breast cancer cell line MDA-MB-231, and use the surface marker CD44 + / CD24 - / low They were sorted according to the standard and divided into breast cancer stem cells (CSC) and breast cancer non-stem cells (NCSC), and then further sorted based on the expression of CXCR6.

[0033] Ⅰ. Sorting of breast cancer stem cells (CSC) and breast cancer non-stem cells (NCSC)

[0034] 1) Sample collection: the human breast cancer cell line MDA-MB-231 is in logarithmic growth phase;

[0035] 2) Centrifugation: 1000rpm / min, 5min, washed twice with PBS;

[0036] 3) Staining: Add 100 μl PBS Buffer, FITC-labeled CD44 (20 μg / ml), PE-labeled CD24 (20 μg / ml), room temperature for 30 minutes in the dark; add 400 μl PBS Buffer before ...

Embodiment 2

[0136] Embodiment 2 The composition of kit of the present invention and the detection of sample to be tested

[0137] 1. RT-PCR detection kit

[0138] 1. The composition of the kit

[0139] Reverse transcription reagents (50 persons):

[0140]

[0141] Amplification and detection reagents (50 persons):

[0142]

[0143] A pair of primers for the target gene CXCR6 (shown in SEQ ID NO.2-3):

[0144] F:5-ATGCCATGACCAGCTTTCACT-3

[0145] R: 5-TTAAGGCAGGCCCTCAGGTA-3

[0146] Primer pair for the housekeeping gene GAPDH (shown in SEQ ID NO.4-5):

[0147] F: 5-GAA GGT GAA GGT CGG AGT C-3

[0148] R: 5-GAAGATGGTGATGGGATTTC-3

[0149] 2. How to use the kit

[0150] (1) Take the sample to be tested, and extract the total RNA of the sample to be tested according to the method in the instruction manual of the RNA extraction kit.

[0151] (2) reverse transcription

[0152] Using the total RNA obtained in step (1) as a template, use the above reverse transcription reagents to...

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Abstract

The invention discloses a screening kit of malignant breast cancer stem cells. The screening kit comprises an optional related reagent for detecting CXCR6 gene expression. The invention also discloses application of the reagent for detecting the CXCR6 gene expression in preparation of a reagent for screening the malignant breast cancer stem cells. The kit provided by the invention is capable of accurately detecting whether a sample to be tested contains the malignant breast cancer stem cells, and provides a reliable index for diagnosis and treatment of clinical breast cancer, and also is bright in clinical application prospect.

Description

technical field [0001] The invention relates to a kit for screening malignant breast cancer stem cells. Background technique [0002] Breast cancer is the malignant tumor with the highest morbidity and mortality among women in the world. Nearly 1.3 million women suffer from breast cancer every year, and more than 400,000 women die of breast cancer metastasis and recurrence. Endocrine hormone imbalance, genetic factors, eating habits, and environmental carcinogens are all key factors in inducing breast cancer, which lead to the occurrence of breast cancer by inducing the activation of oncogenes or inactivation of tumor suppressor genes in human breast cells. With the continuous deepening of basic and clinical research on breast cancer, the treatment methods have also been continuously innovated, and the treatment effect has been increasing year by year. However, with continuous clinical and basic efforts, a considerable number of patients still cannot avoid metastasis and re...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68
Inventor 陈念永彭星辰
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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