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High-transfer kidney clear cell cancer cell system for the Han nationality

A technology for clear cell and renal cell carcinoma, applied in the field of microbiology, can solve the problems of inability to compare the biological characteristics of primary cancer cell lines and cell lines, and the inability to rule out differences

Inactive Publication Date: 2009-08-19
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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Problems solved by technology

However, among the renal cancer cell lines established so far, there is no clear cell renal cell carcinoma cell line that can stably metastasize from Han Chinese metastases, and it is impossible to compare the biological characteristics of primary cancer cell lines and cell lines with a tendency to stably metastasize , and it is impossible to rule out the differences caused by ethnic genetic background and living conditions in the study of kidney cancer in Han Chinese

Method used

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  • High-transfer kidney clear cell cancer cell system for the Han nationality
  • High-transfer kidney clear cell cancer cell system for the Han nationality
  • High-transfer kidney clear cell cancer cell system for the Han nationality

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Obtaining of HiMet-ccRCC cells

[0051] 1) The metastases were excised specimens of spinal metastases from patients with renal clear cell carcinoma in Shanghai Changzheng Hospital. The metastases were taken out under aseptic conditions and placed in a glass plate filled with 10ml of D-Hanks solution, whichever was more than half of the tissue. , to remove necrotic tissue, fat tissue, blood vessels, etc.;

[0052] 2) Soak the tissue block in 15ml1640 medium for 30min;

[0053] 3) Cut metastases into 1mm with ophthalmic scissors 3 Transfer the soaking solution and tissue pieces to a centrifuge tube together, balance the centrifuge tube, shake and wash repeatedly for 2 minutes. 1500rpm, centrifuge for 10min;

[0054] 4) Discard the supernatant, add 7ml of soaking solution to resuspend the tissue pieces, shake and wash repeatedly for 5min, centrifuge at 1500rpm for 10min;

[0055] 5) Discard the supernatant, suspend the tissue pieces with 1ml 1640 complete med...

Embodiment 2

[0060] Animal experiment of highly metastatic renal clear cell carcinoma cell line in Han nationality:

[0061] 1) Take the cell suspension (2×10 6 Cells / 0.2ml) were injected subcutaneously in the back of three 5-week-old NOD-SCID mice (provided by the Tumor Institute of Second Military Medical University). After 90 days, the NOD-SCID mice were sacrificed, and the first round of planting was carried out. The subcutaneous tumor was cut into small tissue pieces with a diameter of 1 mm and inoculated under the renal capsule. After 40 days, a tumor with a diameter of 1 cm was formed in situ.

[0062] 2) The above-mentioned orthotopic tumor material was replanted under the renal capsule of NOD-SCID mice for the second round of planting. After 39 days, the NOD-SCID mice were exhausted, and the lung metastases were removed and replanted under the renal capsule.

[0063] 3) After 27 days, the above lung metastases were removed and replanted under the renal capsule for the third round...

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Abstract

The invention discloses a method for constructing the Hans highly metastatic renal clear cell carcinoma cell line HiMet-ccRCC, CCTCC-C200909 with metastatic phenotype in the Hans renal clear cell carcinoma vertebrae metastasis. The invention adopts culture in vitro to construct a human renal clear cell carcinoma cell line having high metastatic potential with vertebrae metastasis of renal clear cell carcinoma patients as tumor source for constructing the line. Tests show that the inventive cell line has the typical characteristics of tumor cell lines, and is capable of leading NOD-SCID mice to generate stable metastasis and being used as the construction of cell model for the screening preparation detection and prophylaxis of Hans renal cell carcinoma pharmaceuticals.

Description

technical field [0001] The present invention relates to microorganisms, in particular to the establishment of a highly metastatic renal clear cell carcinoma cell line HiMet-ccRCC, CCTCC-C200909, with a metastatic phenotype from a spinal metastases of a Han human clear cell renal cell carcinoma. Background technique [0002] Renal cell carcinoma (RCC) is a common tumor of the urinary system, and the incidence of RCC has shown a significant upward trend in the world in recent years. Renal cell carcinoma has atypical clinical symptoms and is prone to distant metastasis. Nearly 30% of RCC patients are found to have metastases when they are first diagnosed. The average survival period is 6-12 months, and the 2-year survival rate is 10%-20%. Among them, renal clear cell carcinoma is the most common, accounting for 70%-80%. RCC is not sensitive to radiotherapy, chemotherapy, endocrine therapy and other tumor treatment methods. The current relatively stable treatment is surgery, bu...

Claims

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Application Information

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IPC IPC(8): C12N5/08C12Q1/18C12R1/91C12N5/09
Inventor 曹广文谭晓洁林丽萍贺松琴常文军
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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