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
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Embodiment 1
[0042] Example 1, CD73 is the surface marker of renal clear cell carcinoma stem cells
[0043] 1. Obtainment and functional verification of human renal clear cell carcinoma cell line stem cells
[0044] 1. Obtaining stem cells
[0045] 1) Take 786-O cells, a human renal clear cell carcinoma cell line, and culture them in serum-free 1640 supplemented with 10% fetal bovine serum, 100IU / mL penicillin and 100g / mL streptomycin based on 5% CO 2 Incubate at 37°C in an incubator.
[0046]2) After the 786-O cells cultured to the exponential phase in step 1) were digested with trypsin, then treated 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 attachment factors at 5000 cells / well, and microcapsules were formed within 7-10 days. After the microcapsule cells were enriched, the cells were resuspended w...
Embodiment 2
[0081] Example 2. The expression level of CD73 is correlated with the malignant degree (or clinical type) of renal clear cell carcinoma
[0082] Clinical samples of clear cell renal cell carcinoma were obtained from the Second Affiliated Hospital of Lanzhou University. All samples were obtained after partial or total surgical resection of clear cell renal cell carcinoma. The experimental samples were obtained in accordance with the regulations of the 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 samples were acquired from September 2010 to April 2012. All sample mothers did not receive radiation therapy or chemotherapy.
[0083] 1. Clinical classification
[0084] First, the malignancy (or clinical classification) of clear cell renal cell carcinoma of the patient under test is divided into three grades: G1, G2 and G3 according to the following clinical criteria:
[0085]The ...
Embodiment 3
[0095] Example 3. Using the expression level of CD73 to judge the malignant degree (or clinical type) of renal clear cell carcinoma
[0096] 1. Obtaining samples to be tested
[0097] Normal kidney tissues from healthy people, with a sample size of 15; kidney tissues from patients with clear cell renal cell carcinoma, with a sample size of 25. All samples were obtained after partial or total surgical resection of clear cell renal cell carcinoma. The experimental samples were obtained in accordance with Gansu Regulations of the Ethics Committee. All sample mothers did not receive radiation therapy or chemotherapy.
[0098] 2. Detection of the proportion of CD73 positive cells
[0099] Carry out according to the method of step 2 in the embodiment 2, the CD73 positive cell ratio in the renal tissue cells of 15 cases of healthy people is extremely significantly lower than 0.41%, the result of the renal tissue of 25 cases of renal clear cell carcinoma patients is shown in Table 2...
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