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Application of CD73 as stem cell surface marker of renal clear cell carcinoma

A technology of renal clear cell carcinoma and clear cells, which is applied in the application field of CD73 as a surface marker of renal clear cell carcinoma stem cells, which can solve the problems such as stemness analysis of renal clear cell carcinoma that have not yet been seen

Inactive Publication Date: 2013-09-04
INST OF MODERN PHYSICS CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no relevant report on the stemness analysis and clinical classification of clear cell renal cell carcinoma by using CD73

Method used

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  • Application of CD73 as stem cell surface marker of renal clear cell carcinoma
  • Application of CD73 as stem cell surface marker of renal clear cell carcinoma
  • Application of CD73 as stem cell surface marker of renal clear cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1. CD73 is a surface marker of renal clear cell carcinoma stem cells

[0043] 1. Obtaining and functional verification of stem cells of human renal clear cell carcinoma cell line

[0044] 1. Obtaining stem cells

[0045] 1) Take human renal clear cell carcinoma cell line 786-O cells and use serum-free 1640 medium supplemented with 10% fetal bovine serum, 100IU / mL penicillin and 100g / mL streptomycin in 5% CO 2 Incubate at 37°C in an incubator.

[0046] 2) After step 1) the 786-O cells cultured to the exponential phase are digested with trypsin, they are then digested with 10ng / mL fibroblast growth factor, 20ng / mL epidermal growth factor, 5μg / mL insulin and 0.4% fetal bovine serum The protein was resuspended in serum-free 1640 medium, and the cell suspension was seeded into a 6-well plate without adhesion factors at 5000 cells / well, and microcapsules were formed in 7-10 days. After enriching the microencapsulated cells, resuspend the cells in a mild cell separation solut...

Embodiment 2

[0081] Example 2. The expression of CD73 is correlated with the degree of malignancy (or clinical classification) of clear cell renal cell carcinoma

[0082] The clinical samples of clear cell renal cell carcinoma were obtained from the Second Affiliated Hospital of Lanzhou University. All specimens were obtained after partial or complete surgical resection of clear cell renal cell carcinoma. The acquisition of experimental specimens complied with the regulations of Gansu Ethics Committee. Among them, the sample size of normal kidney tissue was 15, and the sample size of renal clear cell carcinoma was 150. The sample acquisition time is from 2010.9 to 2012.4. None of the sample mothers received radiotherapy or chemotherapy.

[0083] 1. Clinical classification

[0084] First, the malignant degree (or clinical classification) of clear cell renal cell carcinoma of the patient to be tested is divided into three levels: G1, G2, and G3 according to the following clinical criteria:

[0085...

Embodiment 3

[0095] Example 3. Using the expression of CD73 to judge the degree of malignancy (or clinical classification) of renal clear cell carcinoma

[0096] 1. Acquisition of samples to be tested

[0097] Normal kidney tissue from healthy people with a sample size of 15; kidney tissue from patients with renal clear cell carcinoma with a sample size of 25. All samples were obtained after partial or complete surgical resection of renal clear cell carcinoma. The experimental samples were obtained in accordance with Gansu Regulations of the ethics committee. None of the sample mothers received radiotherapy or chemotherapy.

[0098] 2. Detection of the proportion of CD73 positive cells

[0099] According to the method of step 2 in Example 2, the results showed that the proportion of CD73-positive cells in the renal tissue cells of 15 healthy people was extremely significantly lower than 0.41%. The results of the renal tissue of 25 patients with renal clear cell carcinoma are shown in Table 2. . ...

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Abstract

The invention discloses an application of CD73 as a stem cell surface marker of renal clear cell carcinoma. The application is that a substance for detecting CD73 can be used for preparing the following products: 1) a product for detecting or screening the stem cells of renal clear cell carcinoma; 2) a product for diagnosis or auxiliary diagnosis of renal clear cell carcinoma; 3) a product for diagnosis or auxiliary diagnosis of the malignancy degree of renal clear cell carcinoma of a patient to be detected; and 4) a product for treating renal clear cell carcinoma by taking CD73 as a target. In-vitro experiments and animal experiments prove that the CD73 can be used as a surface marker of the stem cells of renal clear cell carcinoma; and the result of clinical sample analysis indicates that the proportion of CD73 positive cells is closely related to the clinical classification of renal clear cell carcinoma. The invention provides a new molecular target CD73 to the identification of the stem cells of renal clear cell carcinoma as well as the diagnosis, classification and detection of renal clear cell carcinoma, and provides a simple and efficient method for the prognosis of renal clear cell carcinoma.

Description

Technical field [0001] The present invention relates to the application of CD73 as a surface marker of renal clear cell carcinoma stem cells. Background technique [0002] Renal cancer, also known as renal cell carcinoma, originates from the urinary tubule epithelium. Kidney cancer accounts for about 80%-90% of adult malignant tumors. Renal cell carcinoma is usually divided into 4 types: clear cell type, granular cell type, mixed cell type, and undifferentiated cell type. Among them, clear cell renal cell carcinoma (clear cell renal cell carcinoma, CCRCC) accounts for 70%-80% of renal cancer. The cancer cells are often arranged in sheets, cords, alveoli or tubes, much like renal tubules. Often asymptomatic, or only systemic symptoms such as fever, fatigue, etc., are found when the tumor size increases (JC Cheville, et al. Comparisons of outcome and prognostic features among histologic subtypes of renal cell carcinoma. The American journal of surgical pathology 27( 2003)612-624...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/573G01N33/569A61K39/395A61P35/00A61P13/12
Inventor 周光明王菊芳宋雷叶文凌裴海龙张亚楠丁楠孙放叶才勇高笑菲徐瑚珊
Owner INST OF MODERN PHYSICS CHINESE ACADEMY OF SCI
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